ovalbumin and Food-Hypersensitivity

The Drug Allergy Committee of the Spanish Society of Allergology and Clinical Immunology reviewed the allergenic potential of several substances of food origin that are found in the composition of some drugs. Despite recent legislation on labeling, many labels do not clearly state whether the drug contains raw material (active ingredients, excipient, or other manufacturing intermediate) with an origin in any of the substances in the list of the 14 groups of food allergens that are subject to mandatory declaration. The objective of legislation is that the drug package, the Summary of Product Characteristics, and the patient information leaflet clearly state the food content in order to improve the safety of allergic patients. Therefore, any food or allergen derivative that must be declared should be clearly stated on the drug label. Of all the evaluated products, egg and milk derivatives are the most frequently discussed in literature reviews. The natural or synthetic origin of potentially allergenic substances such as lysozyme, casein, lactose, albumin, phosphatide, and aromatic essences should be clearly stated. Providing this information has 2 clear advantages. First, allergic reactions to drugs in patients with food allergy could be avoided (if the substances have a natural origin). Second, prescription would improve by not restricting drugs containing synthetic substances (which do not usually induce allergic reactions).

Topics: Drug Hypersensitivity; Food Additives; Food Hypersensitivity; Glucosamine; Humans; Lactose; Muramidase; Ovalbumin; Propofol; Spain

Involvement of sub-classes of IgG that are specific for food allergens in anaphylactoid reactions and some manifestations of atopy no longer needs to be shown. Accordingly, sub-classes of IgG specific for ovalbumin (OVA) and beta-lactoglobulin (BLG) were compared in healthy subjects and those who presented with an intolerance or food allergy to OVA and BLG to decide whether a restrictive diet was necessary. The four sub-classes of IgG1, IgG2, IgG3 and IgG4 were isolated in all the groups. IgG4 was highest in the allergic subjects and the IgG sub-class values were modified by the diet differently in each group. Unfortunately, the small number of subjects does not allow the formation of a definite conclusion to this study.

Topics: Antigens; Food Hypersensitivity; Humans; Immunoglobulin G; Lactoglobulins; Ovalbumin

Topics: Aging; Animals; Antigens; Antigens, Bacterial; Cattle; Dietary Proteins; Digestive System; Food Hypersensitivity; Humans; Immunoglobulin A; Immunoglobulin A, Secretory; Immunoglobulin G; Immunoglobulin M; Intestinal Mucosa; Intestines; Lymphocytes; Milk; Milk Proteins; Ovalbumin; Plant Proteins, Dietary; Soybean Proteins

Topics: Antigen-Antibody Complex; Food Hypersensitivity; Humans; Immunity; Immunoglobulin A; Immunoglobulin E; Immunoglobulin G; Immunoglobulin M; Ovalbumin

There is limited understanding of how maternal diet affects breastmilk food allergen concentrations, and whether exposure to allergens through this route influences the development of infant oral tolerance or sensitization.. To investigate how maternal dietary egg ingestion during early lactation influences egg protein (ovalbumin) levels detected in human breastmilk.. In a randomized controlled trial, women were allocated to a dietary group for the first six weeks of lactation: high-egg diet (> 4 eggs per week), low-egg diet (one-three eggs per week) or an egg-free diet. Breastmilk samples were collected at 2, 4 and 6 weeks of lactation for the measurement of ovalbumin. The permeability of the mammary epithelium was assessed by measuring the breastmilk sodium : potassium ratio. Egg-specific IgE and IgG4 were measured in infant plasma at 6 weeks, and prior to the introduction of egg in solids at 16 weeks.. Average maternal egg ingestion was associated with breastmilk ovalbumin concentration. Specifically, for each additional egg ingested per week, there was an average 25% increase in ovalbumin concentration (95% CI: 5-48%, P = 0.01). Breastmilk ovalbumin concentrations were significantly higher in the 'high-egg' group (> 4 eggs per week) compared with the 'egg-free' group (P = 0.04). However, one-third of women had no breastmilk ovalbumin detected. No detectable associations were found between mammary epithelium permeability and breastmilk ovalbumin concentrations. Infant plasma egg-specific IgG4 levels were also positively associated with maternal egg ingestion, with an average 22% (95% CI: 3-45%) increase in infant egg-specific IgG4 levels per additional egg consumed per week (P = 0.02).. Increased maternal egg ingestion is associated with increased breastmilk ovalbumin, and markers of immune tolerance in infants. These results highlight the potential for maternal diet to benefit infant oral tolerance development during lactation.

Topics: Adult; Allergens; Animals; Antibody Specificity; Breast Feeding; Diet; Eggs; Female; Food Hypersensitivity; Humans; Immunoglobulin E; Immunoglobulin G; Infant; Infant, Newborn; Lactation; Male; Milk, Human; Ovalbumin; Pregnancy; Risk Factors

Humoral responses to food antigens may reflect the propensity of a child's immune system to develop tolerance to innocuous antigens. Early nutrition as well as probiotics may influence these immunological responses.. To study the association of humoral responses to early food antigens with the administration of prebiotics and probiotics, with the occurrence of allergy, and with the length of exclusive breastfeeding.. In a randomized double-blind allergy prevention trial in high-risk children, 1018 mothers took probiotics or placebo from the 36th week of gestation, and their newborn infants received probiotics and prebiotics or placebo during 6 months. At 2 and 5 years, we evaluated the cumulative incidence of allergic diseases (food allergy, eczema, asthma, rhinitis) and sensitization (skin prick test ≥3 mm or serum antigen-specific IgE>0.7 kU/L). In 688 infants at age 2, we measured in sera-specific IgA, IgG, IgG1, and IgG4 antibody levels to cow's milk (CM), α-casein (CAS), β-lactoglobulin (BLG), and ovalbumin (OVA) with ELISA, and specific IgE levels to CM and hen's egg with UniCap.. Probiotic treatment (n=342) compared with placebo (n=346) showed no effect on serum food-specific IgA, IgG, IgG1, or IgG4 concentrations at age 2. Atopic children had higher OVA-IgA (P<0.001), OVA-IgG (P=0.001), OVA-IgG1 (P<0.001), and egg-IgE but lower OVA-IgG4/egg-IgE ratio (P<0.001) than non-atopic children. Longer duration of exclusive breastfeeding (≥4 vs. <4 months) was associated with reduced CM- and CAS-specific serum IgA (P<0.001) and IgG levels (P<0.001; P=0.003).. Allergy was associated with more intense IgA and IgG responses to OVA. Breastfeeding depressed humoral responses, whereas prebiotics and probiotics supplementation showed no immunomodulatory effect. The effect of probiotics on allergies is not mediated through food-specific antibody responses. Furthermore, OVA-specific IgA and IgG antibodies may help in assessing the risk for atopy.

Topics: Animals; Antibody Specificity; Breast Feeding; Cattle; Child, Preschool; Double-Blind Method; Female; Food Hypersensitivity; Humans; Hypersensitivity, Immediate; Immunoglobulin A; Immunoglobulin G; Infant; Infant, Newborn; Male; Milk Hypersensitivity; Milk Proteins; Ovalbumin; Prebiotics; Probiotics

Recent Australian and international legislation requires labeling of wines made by using the potentially allergenic food proteins casein, milk, egg white, or isinglass (fish-derived) where "there is a detectable residual processing aid." We investigated whether wines fined using these proteins or non-grape-derived tannins (tree-nut derived) can provoke significant clinical allergic reactions (anaphylaxis) in patients with confirmed immunoglobulin E-mediated relevant food allergy.. A double-blind, placebo-controlled trial was performed to determine whether allergic reactions followed consumption of Australian commercial wines fined using one or more of the legislation-targeted food proteins. In addition, allergenicity of a larger panel of these wines was evaluated by blood basophil activation.. No anaphylaxis was induced by wine consumption. Three mild clinical reactions to protein-fined wine and two mild reactions to unfined wine occurred, but there was no statistically significant difference in reaction parameters between subject groups or between processing aids. No pattern of basophil activation correlated with wine type, processing aid, or subject group.. Wines fined with egg white, isinglass, or non-grape-derived tannins present an extremely low risk of anaphylaxis to fish-, egg-, or peanut-allergic consumers. Although consumption of milk protein-fined wine did not induce anaphylaxis, there were insufficient subjects to determine statistically whether wines fined with milk proteins present a risk to the very rare milk-allergic consumers. In summary, the observed lack of anaphylaxis and basophil activation induced by wines made using the legislation-targeted food proteins according to good manufacturing practice suggests negligible residual food allergens in these wines.

Topics: Adult; Allergens; Anaphylaxis; Arachis; Basophils; Caseins; Double-Blind Method; Female; Food Contamination; Food Handling; Food Hypersensitivity; Gelatin; Humans; Male; Middle Aged; Ovalbumin; Tannins; Wine; Young Adult

While immediate-type clinical reactions to food can quite easily be identified by history or measurement of specific IgE in combination with positive oral food challenges, the evaluation of food allergy in the absence of immediate clinical reactions still presents diagnostic difficulties--particularly in children with atopic dermatitis. The objective of this study was to evaluate the diagnostic value of the atopy patch test (APT) with regard to late-phase reactions observed in double-blind, placebo-controlled food challenges with cow's milk, hen's egg, wheat, and soybean.. We investigated 75 children (median age 2.1 years) with suspected food allergy by double-blind, placebo-controlled food challenges, specific IgE in serum, skin prick test, and APT. Of the subjects, 69/75 suffered from atopic dermatitis.. Of 209 oral challenges, 133 were performed with allergen and 76 with placebo. We assessed 77/133 allergen and 2/76 placebo challenges as positive. In 66 of 77 (86%) positive oral challenges, specific IgE in serum to the corresponding allergen was positive; in 64/77 (83%) the skin prick test, and in 42/77 (55%) the APT was positive. While immediate-type reactions were associated with positive skin prick test and proof of specific IgE in serum, late-phase clinical reactions were associated with a positive APT (sensitivity 76%, specificity 95%).. The APT seems to be a valuable additional tool in the diagnostic work-up of food allergy in children with atopic dermatitis - especially with regard to late-phase clinical reactions. The APT may help to prevent unnecessary restrictive diets which may be the consequence of misjudging late reactions by clinical assessment alone.

Topics: Animals; Child; Child, Preschool; Dermatitis, Atopic; Double-Blind Method; Female; Food Hypersensitivity; Glycine max; Humans; Immunoglobulin E; Infant; Intradermal Tests; Male; Milk; Ovalbumin; Patch Tests; Predictive Value of Tests; Sensitivity and Specificity; Triticum

We occasionally see egg-allergic children who develop contact urticaria to hen's egg despite the absence of the overt symptoms on ingestion. The mechanisms remain to be elucidated.. Twenty-one subjects with positive reactions to 20-min patch tests for egg-white antigens were divided into subgroups with positive (n = 10) and negative (n = 11) results to oral challenge tests by the same antigens. We measured IgE antibody for egg white and its components, and IgE-binding activities to digestive enzyme-treated ovomucoid by RAST inhibition.. There were no significant differences in IgE antibody titers to egg white (positive vs negative: 30.3% vs 15.3%, P=0.130), ovomucoid (21.5% vs 10.2%, P= 0.078), ovotransferrin (9.9% vs 3.7%, P = 0.105), and lysozyme (3.4% vs 2.9%, P=0.944), except ovalbumin (16.8% vs 5.6%, P=0.024), between the positive and negative subjects in the provocation tests. In contrast, the concentration (1.93 microg/ml) of pepsin-treated ovomucoid needed for 50% RAST inhibition in the challenge-positive subjects was significantly (P=0.0003) lower than that (114.9 microg/ml) of negative subjects. Similar but less significant differences were obtained when ovomucoid fragments treated with chymotrypsin (0.91 microg/ml vs 6.86 microg/ml, P=0.014) and trypsin (0.75 microg/ml vs 4.67 microg/ml, P= 0.041) were used as inhibitors.. We suggest that IgE antibodies from subjects showing contact urticaria despite the absence of reactions to the ingestion of egg white recognize the epitope(s) unstable to digestive enzymes.

Topics: Administration, Oral; Allergens; Animals; Child; Child, Preschool; Chymotrypsin; Conalbumin; Double-Blind Method; Egg White; Eggs; Female; Food Hypersensitivity; Humans; Immunoglobulin E; Infant; Male; Muramidase; Ovalbumin; Ovomucin; Pepsin A; Placebos; Radioallergosorbent Test; Trypsin; Urticaria

No egg white products have been clearly proven to be hypoallergenic. The role of egg white proteins in allergic reactions to eggs is still debatable.. This study was designed to determine the importance of ovomucoid, an egg white protein, in the development of allergies to egg white.. We performed a double-blind, placebo-controlled food challenge in subjects with high levels of IgE antibodies for egg white to compare the allergenicities of heated and ovomucoid-depleted egg white, freeze-dried egg white, and heated egg white. Levels of IgE antibodies for egg white, ovomucoid, ovalbumin, ovotransferrin, and lysozyme were measured in serum by RAST.. Twenty-one of 38 subjects with positive challenge responses to freeze-dried egg white had negative challenge responses to heated egg white, whereas 16 of 17 subjects (94.1%) with positive responses to heated egg white did not respond to the heated and ovomucoid-depleted egg white challenge. The subjects with positive challenge responses to freeze-dried egg white tended to have higher IgE antibody values to ovomucoid than those with negative responses. IgE antibody levels to ovomucoid were significantly higher in subjects with positive responses to a challenge with heated egg white than in those with no response. There were no significant differences in the levels of IgE antibodies to the other proteins, except ovomucoid, in the negative-response and positive-response groups in challenge tests with freeze-dried and heated egg white.. The heated and ovomucoid-depleted egg white preparation was less allergenic than heated or freeze-dried preparations. Ovomucoid has a more important role in the pathogenesis of allergic reactions to egg white than other proteins in egg white.

Topics: Allergens; Antibodies, Anti-Idiotypic; Antibody Specificity; Child; Child, Preschool; Conalbumin; Double-Blind Method; Egg White; Electrophoresis, Polyacrylamide Gel; Female; Food Hypersensitivity; Hot Temperature; Humans; Infant; Male; Muramidase; Ovalbumin; Ovomucin; Placebos; Sodium Dodecyl Sulfate

Food antigens transferred into breast milk sometimes cause an allergic reaction in exclusively breast-fed infants. This study will show whether the intake of a whey hydrolysate formula for lactating women (MOM HA) can reduce the appearance of food antigens in breast milk. Lactating women in the MOM group (n = 12) consumed MOM HA as a substitute for cow's milk and those in the COW group (n = 13) consumed cow's milk for more than 4 months. After the ingestion of 200 mL of MOM HA and cow's milk by the women in the MOM and COW groups, respectively, the first breast milk samples were obtained and beta-lactoglobulin was measured using enzyme-linked immunosorbent assay. The number of subjects with detectable beta-lactoglobulin (> 0.1 ng/mL) in the MOM group was two (17%), which was significantly less than that in the COW group (11 subjects, 85%, p < 0.01). The level of beta-lactoglobulin was also lower in the MOM group than the COW group (p < 0.01). Subsequently, the women in the MOM group consumed cow's milk and those in the COW group consumed MOM HA for one week; then a second sampling was performed. beta-Lactoglobulin was detected in three (25%) and 8 subjects (62%) in the MOM and COW groups, respectively. The level of beta-lactoglobulin was still lower in the MOM group (p < 0.05). The consumption of whey hydrolysate formula by lactating women over a considerable time reduces the transfer of beta-lactoglobulin into their breast milk, and the low level can be maintained even after inadvertent ingestion of cow's milk.

Topics: Animals; Cattle; Female; Food Hypersensitivity; Humans; Hydrolysis; Infant, Newborn; Lactation; Lactoglobulins; Milk Proteins; Milk, Human; Ovalbumin; Whey Proteins

The causal relation between egg allergy and cytokines derived from lymphocytes is unknown.. Interferon-gamma (IFN-gamma) and interleukin-4 (IL-4) production of ovalbumin-stimulated and interleukin-2 (IL-2)-stimulated peripheral blood mononuclear cells from egg-sensitive patients was investigated and compared with that of stimulated peripheral blood mononuclear cells from nonatopic healthy children.. Peripheral blood mononuclear cells from egg-sensitive patients and nonatopic healthy children were cultured with ovalbumin and IL-2 for seven days. The IFN-gamma and IL-4 concentrations in culture supernatants of the peripheral blood mononuclear cells were investigated.. The levels of IFN-gamma production of only IL-2-stimulated or both ovalbumin-stimulated and IL-2-stimulated peripheral blood mononuclear cells from egg-sensitive patients with atopic dermatitis was significantly higher than that of healthy children and that of egg-sensitive patients with immediate allergic symptoms.. Increased IFN-gamma production by lymphocytes after IL-2 and antigen stimulation has important implications for the mechanism of food-sensitive atopic dermatitis.

Topics: Cell-Free System; Cells, Cultured; Child; Child, Preschool; Double-Blind Method; Eggs; Female; Food Hypersensitivity; Humans; Immunoglobulin E; Infant; Interferon-gamma; Interleukin-4; Lymphocyte Activation; Male; Ovalbumin; Radioallergosorbent Test

Experimental studies have shown that ketotifen fumarate inhibits reaginic antibody-mediated hypersensitivity reactions. In this study, the proliferative responses of peripheral blood mononuclear cells (PBMCs) to ovalbumin (OA) in children with atopic dermatitis (AD), who are sensitive to hen's egg, were significantly higher than those of healthy children. The proliferative responses of PBMCs to OA were dose-dependently inhibited by ketotifen in patients with hen's egg-sensitive AD. Moreover, the inhibition resulted from the effects of ketotifen on T cells. In contrast, the proliferative responses of PBMCs to phytohemagglutinin and tetanus toxoid were not inhibited by ketotifen. These results suggest that ketotifen inhibits food antigen-specific proliferative responses of PBMCs in patients with food-sensitive AD.

Topics: Child, Preschool; Depression, Chemical; Dermatitis, Atopic; Double-Blind Method; Epitopes; Food Hypersensitivity; Humans; Infant; Ketotifen; Lymphocyte Activation; Ovalbumin; Serum Albumin, Bovine

N(3',4'-dimethoxycinnamoyl) anthranilic acid (Tranilast) inhibits antibody-mediated hypersensitivity reactions, and is an effective drug for patients with bronchial asthma or allergic rhinitis. Interferon-gamma (IFN-gamma) production of ovalbumin (OA)-stimulated peripheral blood mononuclear cells (PBMCs) from hen's egg-sensitive patients with atopic dermatitis (AD) was significantly higher than those of healthy controls. Tranilast inhibited this IFN-gamma production. Moreover, interleukin-2 (IL-2) production of OA-stimulated PBMCs from hen's egg-sensitive patients with AD was also inhibited by Tranilast. Our results suggest that Tranilast can be used to the patients with food sensitive AD.

Topics: Anti-Allergic Agents; Cells, Cultured; Child, Preschool; Dermatitis, Atopic; Double-Blind Method; Food Hypersensitivity; Humans; Infant; Interferon-gamma; Interleukin-2; Lymphocytes; ortho-Aminobenzoates; Ovalbumin

The timing of onset of allergic symptoms in a double-blind, placebo-controlled food challenge (DBPCFC) was combined with a radioallergosorbent test (RAST) and evaluation of proliferative responses of peripheral blood mononuclear cells (PBMCs) in 27 atopic dermatitis patients with hen's egg allergy. Of the 27 patients, 11 showed reactions within 2 h (immediate group), and 16 showed reactions after more than 2 h (nonimmediate group) after ingestion of hen's egg. The RAST scores for hen's egg in the immediate group were significantly (p < 0.01) higher than those in the nonimmediate group, while the proliferative responses of PBMCs to ovalbumin in the nonimmediate group were significantly (p < 0.01) higher than those in the immediate group. These results suggest that the RAST values are related to immediate allergic symptoms in DBPCFC and the proliferative responses of PBMCs are related to nonimmediate allergic symptoms in DBPCFC. The timing of onset of allergic symptoms in DBPCFC will render DBPCFC more useful in the diagnosis of food allergy, including the late-onset reactions to foods.

Topics: Adolescent; Cells, Cultured; Child; Child, Preschool; Dermatitis, Atopic; DNA; Double-Blind Method; Eggs; Evaluation Studies as Topic; Food Hypersensitivity; Humans; Infant; Lymphocyte Activation; Lymphocytes; Ovalbumin; Radioallergosorbent Test; Time Factors

Serum antibodies to cow milk proteins and ovalbumin were measured quantitatively. Food hypersensitivity of the immediate type was determined to be present or absent by skin tests and double-blind food challenges. Elevated levels of antibodies to milk proteins in sera characteristic of infants fed cow milk were found to decline with age, so that sera from children who were 6 to 15 years of age (inclusive), not hypersensitive to food, had significantly lower levels than the infants. In contrast, sera from age-matched children, who were shown to have hypersensitivity to some food, were found to have levels of antibodies to milk proteins as elevated as in infancy. Hypersensitivity was not necessarily to milk but often to some other food. This persistence of greater antibody production to milk throughout childhood in those hypersensitive to some food indicates a fundamental difference from those without hypersensitivity to food, either in permeability, in immunological reactivity of the gut or in development of immunological unresponsiveness. Implications for pathogenesis of clinical disorders are discussed.

Topics: Adolescent; Allergens; Alternaria; Animals; Antibodies; Binding Sites, Antibody; Cattle; Child; Child, Preschool; Clinical Trials as Topic; Food Hypersensitivity; Histamine; Humans; Hypersensitivity, Immediate; Infant; Milk Proteins; Ovalbumin; Plants; Skin Tests; Time Factors

Food allergy diagnosis and management causes a number of social and emotional challenges for individuals with food allergies and their caregivers. This has led to increased interest in developing approaches to accurately predict food allergy diagnosis, severity of food allergic reactions, and treatment outcomes. However, the utility of these approaches is somewhat conflicting.. We sought to develop and utilize a murine model that mimics the disease course of food allergy diagnosis and treatment in humans and to identify biomarkers that predict reactivity during food challenge (FC) and responsiveness during oral immunotherapy (OIT) and how these outcomes are modified by genetics.. Skin-sensitized intestinal IL-9 transgenic (IL9Tg) and IL9Tg mice backcrossed onto the IL-4Rα. Subcutaneous sensitization and a single intragastric allergen challenge of egg antigen to BALB/c IL9Tg mice and Il4ra. Collectively, these data indicate that single nucleotide polymorphisms in IL-4Rα can alter clinical symptoms of food allergic reactions, severity, and reactive dose during FC and OIT, and that severity of first reaction can predict the likelihood of reaction during FC in mice with IL-4Rα gain of function.

Topics: Administration, Oral; Allergens; Animals; Biomarkers; Desensitization, Immunologic; Food Hypersensitivity; Humans; Immunotherapy; Mice; Mice, Transgenic; Ovalbumin

The incidence and prevalence of food allergy have sharply risen over the past several decades. Oral administration of probiotic stains has been proven as a safe and effective method to control food allergy. In this study, it aims to comprehensively investigate the anti-allergic effect of Lactobacillus plantarum JC7.. OVA-sensitised mice showed mitigation of respiratory manifestations, alleviation of lung inflammation and congestion, and the presence of an intact intestinal villus structure. Furthermore, OVA-specific immunoglobulin E (IgE), OVA-specific-IgG1, and plasma histamine levels were declined in mice treated with L. plantarum JC7 than in OVA-sensitised mice. In addition, interferon-γ (IFN-γ) and interleukin 10 (IL-10) levels were significantly increased, while IL-4 and IL-17A levels were clearly decreased in mice that had undergone oral administration of L. plantarum JC7, compared with OVA-sensitised mice. These findings indicated imbalances of T helper cell type 1 (Th1)/Th2 and regulatory T cells (Treg)/Th17, which were confirmed by quantitative polymerase chain reaction (PCR). Western blotting demonstrated that the expression levels of phosphorylated IκBα and nuclear factor kappa B p65 were significantly increased in OVA-sensitised mice, but these changes were partly reversed after treatment with L. plantarum JC7. Oral administration of L. plantarum JC7 increased the richness, diversity, and evenness of cecum microbiota, characterised by higher Bacteroidetes abundance and lower Firmicutes abundance. Additionally, the intestinal microbial community composition was significantly altered in the OVA-sensitised group, indicating a disordered intestinal microbiota that was restored by the oral administration of L. plantarum JC7.. Overall, L. plantarum JC7 can prevent food allergy by rectifying Th1/Th2 and Treg/Th17 imbalances, combined with modifications of disordered intestinal microbiota.

Topics: Administration, Oral; Animals; Cytokines; Disease Models, Animal; Food Hypersensitivity; Gastrointestinal Microbiome; Lactobacillus plantarum; Mice; Mice, Inbred BALB C; NF-kappa B; NF-KappaB Inhibitor alpha; Ovalbumin; RNA, Ribosomal, 16S

Epidemiologic studies suggest a link between the incidence of food allergy and the consumption of dietary advanced glycation end-products (AGEs). However, the pathogenic role of dietary AGEs in food allergy is largely unknown. This study aims to investigate the effect of allergen-specific and non-specific AGEs on the allergenic manifestation of ovalbumin (OVA), a typical food allergen in vivo.. OVA is glycated by methylglyoxal to prepare allergen-specific AGEs (i.e., OVA-AGE), and a standard AIN-93G diet is heated to obtain allergen-non-specific AGEs. A BALB/c mouse model orally sensitizes to OVA with different forms of AGEs is established and the outcomes are measured as clinical signs, specific antibodies, type-2/type-2 cytokines, immune cell subpopulations, intestinal barrier function, and gut microbiota (GM) composition. The OVA-AGE which has a lower immunoglobulin E (IgE)-binding level in vitro does not reduce the allergenicity of OVA but promotes a stronger T helper 2 cells (Th2)-response than native OVA in vivo. Both forms of AGEs up-regulate the expression of splenic RAGE and aggravate the destruction of gut barrier and GM dysbiosis, especially when exposes to non-relevant AGEs.. This study highlights the role of dietary AGEs in food allergy and helps to understand the biological consequences of immune-toxic compounds in modern diet.

Topics: Allergens; Animals; Cytokines; Disease Models, Animal; Food Hypersensitivity; Glycation End Products, Advanced; Mice; Mice, Inbred BALB C; Ovalbumin

To test the effect of two dietary antioxidants: butylated hydroxytoluene (BHT) and 3-hydroxytyrosol (3-HT) in experimental food allergy.. BALB/c mice maintained on control diet or diet with BHT or 3-HT were sensitized with ovalbumin (OVA) or saline through transdermal exposure. Plasma OVA-specific IgE (OVA-IgE) and IgG1 (OVA-IgG1) antibody levels were determined using ELISA. Sensitized mice were challenged by oral gavage with OVA. Rectal temperature (RT) was measured before and after challenge. Mast cell degranulation was quantified by measuring the plasma levels of mouse mucosal mast cell protease-1 (mMCP-1). Flow cytometry was carried out to evaluate the percentage Th2 cells from the spleen.. Mice on either a 3-HT or BHT diet showed a significantly decreased IgE response to OVA sensitization and less severe anaphylaxis, as evidenced by a diminished drop in body temperature, attenuated clinical signs, a more rapid recovery and decreased mast cell degranulation (as determined by lower plasma mMCP-1 levels).. The present study indicates two dietary antioxidants: BHT and 3-HT may be protective against experimental food allergy. These results suggest 3-HT and BHT could potentially be useful for prevention of food allergy.

Topics: Animals; Antioxidants; Butylated Hydroxytoluene; Disease Models, Animal; Food Hypersensitivity; Immunoglobulin E; Immunoglobulin G; Mast Cells; Mice; Mice, Inbred BALB C; Ovalbumin

Rosmarinic acid (RA) has been proven to exert antianaphylaxis in atopic dermatitis, asthma, and allergic rhinitis. The aim of this study was to determine the hepatoprotective effects of RA on ovalbumin (OVA) challenge-induced intestinal allergy. The results exhibited that RA could relieve anaphylactic symptoms, decrease diarrhea, and prevent hypothermia in allergic mice. Moreover, the elevation of OVA specific IgE (OVA-sIgE), histamine, and mouse mast cell proteinases (mMCP-1) in the serum of OVA challenged mice were remarkably inhibited by RA. OVA challenge resulted in notable increases in serum alanine aminotransferase (ALT), aspartate aminotransferase (AST) activities, liver malondialdehyde (MDA) and nitic oxide (NO) levels, and a remarkable decrease in liver superoxide dismutase (SOD) activity and glutathione (GSH) level. RA treatments succeeded in improving these biochemical parameters and promote the redox homeostasis. Cytokine expression evaluation showed that RA effectively enhanced the expression of anti-inflammatory cytokines (IL-10 and FOXP-3) in the liver of OVA-challenged mice. Meanwhile, the elevation of pro-inflammatory cytokines (TNF-α, IL-4, IL-6, mMCP-1, and iNOS) were remarkably inhibited by RA. These findings suggest that RA possesses hepatoprotective effects on OVA challenge-induced liver injury. The anti-oxidative and anti-inflammatory activities of RA potentially play vital roles in this process.

Topics: Animals; Anti-Inflammatory Agents; Cytokines; Disease Models, Animal; Food Hypersensitivity; Mice; Mice, Inbred BALB C; Ovalbumin; Rosmarinic Acid

Food allergy has become a world recognized public health problem due to its versatility and lack of efficacious methods for its treatment. Probiotics supplement is a potential way to prevent food allergy.. In this study, potential strains are screen out by peripheral blood mononuclear cells (PBMCs), and their abilities of alleviating food allergy are examined using a mouse model induced by ovalbumin (OVA). The results show that six strains increase ratio of interferon-γ (IFN-γ)/interleukin (IL)-4 secreted by PBMCs with good abilities in intestinal adhesion and gastrointestinal tolerance. Oral administration of Bifidobacterium animalis KV9 (KV9) and Lactobacillus vaginalis FN3 (FN3) attenuates allergic responses in allergy mice, including allergic symptoms, mast cells aggregation and activity, serum OVA-special-immunoglobulins E (OVA-sIgE) production. KV9 and FN3 upregulate the production of IFN-γ/IL-4 in splenocytes, increase the genes and proteins expression of toll-like receptor 4 (TLR4), myeloid differentiation primary response 88 (Myd88) and interferon regulatory factor (IRF)-1 in allergic mice spleen, and decrease the IRF-4.. The study demonstrates that KV9 and FN3 possess anti-allergic activities via activation of TLR4 pathway and modulating the expression of IRF-1 and IRF-4 which leads to T helper type 1 (Th1)/T helper type 2 (Th2) cell immunology balance.

Topics: Allergens; Animals; Cytokines; Food Hypersensitivity; Interferon-gamma; Leukocytes, Mononuclear; Mice; Mice, Inbred BALB C; Ovalbumin; Probiotics; Signal Transduction; Th1 Cells; Th2 Cells; Toll-Like Receptor 4

Various studies have reported relationships between salt intake and diseases, such as hypertension, cardiovascular disease, stroke, gastric cancer, and bronchial asthma. However, no reports exist on the relationship between salt intake and food allergies. In this study, we investigated the effect of continuous ingestion of sodium chloride (NaCl) on allergy symptoms using a mouse model of food allergy. BALB/c mice were divided into four groups of 6-8 animals each. The control-water group (CW) and sensitization-water group (SW) groups were provided free access to water, and the control-1% NaCl group (CS) and sensitization-1% NaCl group (SS) groups were provided a 1% NaCl solution. The SW and SS groups were sensitized with 50 µg ovalbumin (OVA) at 2 timepoints by intraperitoneal injection. After oral administration of OVA, anaphylactic response was measured and blood was collected. The mice were sacrificed, and serum levels of OVA and anti-OVA immunoglobulin (Ig)E and IgG1 were measured by enzyme-linked immunosorbent assays. The sodium ion (Na

Topics: Allergens; Animals; Disease Models, Animal; Eating; Food Hypersensitivity; Mice; Mice, Inbred BALB C; Ovalbumin; Sodium Chloride; Sodium Chloride, Dietary

Food allergy is recognized as a global medical problem with increasing prevalence in recent years. Currently, the treatment of food allergy mainly involves avoidance of allergens and allergen-specific immunotherapy. Barring the spontaneous resolution of food allergy during the growth process, this disease is difficult to treat fundamentally. In recent years, the use of functional food ingredients derived from natural products has been attracting attention for their prophylactic use in food allergy. Theaflavins, i.e., black tea polyphenols, are potent antioxidants that have inhibitory effects on a variety of diseases. However, little is known about the preventive effect of theaflavins on food allergy. In this study, we designed a mouse model of food allergy and examined the effect of theaflavins using the severity of diarrhea, a symptom of food allergy, as an indicator. The administration of a black tea extract rich in theaflavins or theaflavin 1 (subgroup of theaflavins) to mice reduced the severity of diarrhea when compared with a normal diet. A reduction in malondialdehyde levels, a key marker of lipid peroxidation, was also observed. Overall, these data suggest that theaflavins may potentially inhibit food allergy by alleviating oxidative stress in the colon and can be a potential food material for prevention of food allergy.

Topics: Animals; Antioxidants; Food Hypersensitivity; Mice; Ovalbumin; Polyphenols; Tea

A major route of sensitization to food allergen is through an impaired skin barrier. IL-33 and thymic stromal lymphopoietin (TSLP) have both been implicated in epicutaneous sensitization and food allergy, albeit in different murine models.. We assessed the respective contributions of TSLP and IL-33 to the development of atopic dermatitis (AD) and subsequent food allergy in TSLP and IL-33 receptor (ST2)-deficient mice using an AD model that does not require tape stripping.. ASP and/or OVA patched, but not OVA-alone patched, BALB/cJ mice developed an AD-like skin phenotype. However, epicutaneous OVA sensitization occurred in OVA patched mice and was decreased in ST2. Epicutaneous sensitization to food allergen and development of food allergy can occur without skin inflammation and is partly mediated by TSLP, suggesting that prophylactic targeting of TSLP may be useful in mitigating the development of AD and food allergy early in life in at-risk infants.

Topics: Allergens; Animals; Cytokines; Dermatitis, Atopic; Disease Models, Animal; Food Hypersensitivity; Interleukin-1 Receptor-Like 1 Protein; Interleukin-33; Mice; Mice, Inbred BALB C; Ovalbumin; Thymic Stromal Lymphopoietin

Acrylamide (AA) forms during the thermal processing of food, but adversely affects human health. As the consumption of heat-processed foods increases, the potentially harmful effect of AA on food allergies needs to be clarified. Here, we investigated how AA affects the allergenicity of OVA in vivo using a mouse model of orally induced OVA allergy. AA enhanced OVA-induced food allergic response by increasing IgE, IgG, IgG1, histamine, and MCP-1. AA promoted the Th2 cell response to modulate the imbalance in Th1/Th2. Furthermore, AA reduced the expression of intestinal tight junction proteins, and disrupted the permeability of the intestine, which impaired the intestinal epithelial barrier, resulting in more OVA crossing it. These actions aggravated the allergic reaction of OVA. In conclusion, this study confirmed the potentially harmful effect of AA on food allergy.

Topics: Acrylamides; Allergens; Animals; Cytokines; Disease Models, Animal; Food Hypersensitivity; Humans; Immunoglobulin E; Intestines; Mice; Mice, Inbred BALB C; Ovalbumin

Food allergies (FAs) are caused by a failure of the immune system to regulate oral tolerance (OT). The use of soap containing hydrolyzed wheat overrides acquired OT to wheat through skin exposure. However, in mouse models, the experimental OT is robust, suggesting that acquired OT to allergens prevents the development of FAs. We aimed to analyze the mechanisms and developed a mouse model of FA that overrides acquired OT via skin exposure. Three murine FA models (intraperitoneal [IP], epicutaneous [EC], and intradermal [ID]) were compared to evaluate if allergies to ovalbumin (OVA) that had been previously tolerated orally could be induced. In the ID model, OT was overridden, and allergic reactions of severe anaphylaxis were developed. To analyze this effect in the ID model, we measured the migration of dendritic cells (DCs) into lymph nodes. The induction of OT promoted the migration of CD103

Topics: Allergens; Anaphylaxis; Animals; Disease Models, Animal; Food Hypersensitivity; Mice; Mice, Inbred BALB C; Ovalbumin; Skin

Objective To investigate the preventive therapeutic effect and possible mechanism of single chain variable fragments chimeric protein (SD) of ovalbumin epitopes internalizing receptor DEC-205 antibody on food allergy in mice. Methods Mice were randomly divided to five groups (control, PBS, scFv DEC 100 μg, SD 50 μg, SD 100 μg) and treated for 24 hours before OVA administration. After challenge, the serum level of OVA-specific IgE, IgG1, IgG2a and IL-4 were detected by ELISA. Infiltration of eosinophils and mast cells in the jejunum was observed by HE staining and toluidine blue staining respectively. The bone marrow of tibia and femur was isolated and cultured to obtain immature dendritic cells(BMDCs), which were further treated with LPS (10 ng/mL), TSLP (50 ng/mL), scFv DEC protein (1000 ng/mL) and SD protein (10,100,1000)ng/mL for 24 hours, and the IL-10 level of supernatant was assayed by ELISA. Results Compared with PBS group, the number of SD-treated mice with diarrhea was markedly reduced. The difference in rectal temperature and the levels of serum OVA-specific IgE, IgG1, IgG2a and IL-4 decreased significantly after prophylactic administration of SD; The number of eosinophils and mast cells in jejunum also decreased significantly while the IL-10 level in the supernatant of BMDCs increased significantly after SD intervention. Conclusion SD mitigates experimental FA response by fosters the immune tolerance property of dendritic cells.

Topics: Animals; Disease Models, Animal; Epitopes; Food Hypersensitivity; Immunoglobulin E; Immunoglobulin G; Interleukin-10; Interleukin-4; Mice; Mice, Inbred BALB C; Ovalbumin; Recombinant Fusion Proteins; Single-Chain Antibodies

Patients with food allergy often suffer from atopic dermatitis, in which. The non-tape-stripped skins of wild-type,. Stimulation with δ-toxin induced the release of IL-1α, but not IL-33, in murine keratinocytes. Epicutaneous treatment with OVA and δ-toxin induced the local production of IL-1α. This treatment induced the translocation of OVA-loaded cDC2 from skin to draining LN and OVA-specific IgE production, independently of mast cells and ST2. This resulted in OVA-administered food allergic responses. In these models, pretreatment with anti-IL-1α antibody inhibited the cDC2 activation and OVA-specific IgE production, thereby dampening food allergic responses.. Even without tape stripping, δ-toxin present on skin enhances epicutaneous sensitization to food allergen in an IL-1α-dependent manner, thereby promoting the development of food allergy.

Topics: Animals; Dermatitis, Atopic; Disease Models, Animal; Exotoxins; Food Hypersensitivity; Immunoglobulin E; Interleukin-1 Receptor-Like 1 Protein; Mice; Ovalbumin; Staphylococcus aureus

Obesity and type 2 diabetes (T2D) have been found to be associated with abnormalities in several organs, including the intestine. These conditions can lead to changes in gut homeostasis, compromising tolerance to luminal antigens and increasing susceptibility to food allergies. The underlying mechanisms for this phenomenon are not yet fully understood. In this study, we investigated changes in the intestinal mucosa of diet-induced obese mice and found that they exhibited increased gut permeability and reduced Treg cells frequency. Upon oral treatment with ovalbumin (OVA), obese mice failed to develop oral tolerance. However, hyperglycemia treatment improved intestinal permeability and oral tolerance induction in mice. Furthermore, we observed that obese mice exhibited a more severe food allergy to OVA, and this allergy was alleviated after treatment with a hypoglycemic drug. Importantly, our findings were translated to obese humans. Individuals with T2D had higher serum IgE levels and downregulated genes related to gut homeostasis. Taken together, our results suggest that obesity-induced hyperglycemia can lead to a failure in oral tolerance and to exacerbation of food allergy. These findings shed light on the mechanisms underlying the relationship among obesity, T2D, and gut mucosal immunity, which could inform the development of new therapeutic approaches.

Topics: Administration, Oral; Allergens; Animals; Diabetes Mellitus, Type 2; Food Hypersensitivity; Humans; Immune Tolerance; Mice; Mice, Inbred BALB C; Mice, Obese; Obesity; Ovalbumin

Egg is one of the eight major food allergens, and ovalbumin (OVA) is the most abundant allergenic protein in eggs. In this study, the effects of pulsed electric field (PEF)-assisted Alcalase hydrolysis on the spatial conformation and potential allergenicity of OVA were studied, and the mechanism of its inhibiting allergic reactions effect was revealed. PEF-assisted Alcalase hydrolysis increased the degree of hydrolysis, surface hydrophobicity, and free sulfhydryl group content. Moreover, the reduction in the α-helix content, fluorescence intensity, and disulfide bond content suggested that PEF promoted the OVA hydrolysis by Alcalase. Additionally, enzyme-linked immunosorbent assay data indicated that PEF-assisted Alcalase hydrolysis hindered OVA binding to immunoglobulins E and G1. Finally, based on bioinformatics combined with mass spectrometry, PEF-assisted Alcalase reduced OVA-induced allergic reactions by destroying epitopes in OVA. Overall, PEF technology further destroyed the epitopes of allergens by targeting the binding sites of substrates and enzymes to improve the affinity of enzymes and substrates, reducing allergic reactions.

Topics: Allergens; Epitopes; Food Hypersensitivity; Humans; Ovalbumin; Subtilisins

Previous research studies have shown that sulfated polysaccharides can inhibit food allergy, but the detailed mechanism remains largely unknown. In this study, RBL-2H3 cells were used to compare the anti-allergic activities of four sulfated polysaccharides, and an ovalbumin (OVA)-sensitized allergic mouse experiment was used to explore their desensitization effect, with regard to the alteration in gut microbiota and immune cell differentiation. Compared with the shark, bovine and porcine chondroitin sulfate, sea cucumber chondroitin sulfate (SCCS) significantly inhibited the degranulation of RBL-2H3 cells. SCCS reduced allergic symptoms and protected the jejunum from injury in mice. Furthermore, SCCS increased the relative abundance of

Topics: Animals; Anti-Allergic Agents; Cattle; Cell Differentiation; Chondroitin Sulfates; Disease Models, Animal; Food Hypersensitivity; Gastrointestinal Microbiome; Mice; Mice, Inbred BALB C; Ovalbumin; Polysaccharides; Sea Cucumbers; T-Lymphocytes, Regulatory

Food allergies are an exceptional response of the immune system caused by the ingestion of specific foods. The main foods responsible for allergic reactions are milk, eggs, seafood, soy, peanuts, tree nuts, wheat, and their derived products. Chicken egg ovalbumin (OVA), a common allergen molecule, is often used for the clarification process of wine. Traces of OVA remain in the wine during the fining process, and they can cause significant allergic reactions in sensitive consumers. Consequently, the European Food Safety Authority (EFSA) and the American Food and Drug Administration (FDA) have shown the risks for allergic people to assume allergenic foods and food ingredients, including eggs. Commonly, OVA detection requires sophisticated and time-consuming analytical techniques. Intending to develop a faster assay, we designed a proof-of-concept non-Faradaic impedimetric immunosensor for monitoring the presence of OVA in wine. Polyclonal antibodies anti-OVA were covalently immobilised onto an 11-mercaptoundecanoic-acid (11-MUA)-modified gold surface. The developed immunosensor was able to detect OVA in diluted white wine without the need for an external probe or any pre-treatment step with a sensitivity of 0.20 µg/mL, complying with the limit established by the resolution OIV/COMEX 502-2012 for the quantification of allergens in wine.

Topics: Allergens; Biosensing Techniques; Electric Impedance; Food Hypersensitivity; Humans; Immunoassay; Ovalbumin; Wine

Although an animal model of food allergy has been used to investigate its progression mechanism, most researcher could not assess its symptoms for long especially under dark environment. We assessed the behavioral changes of food allergic mice using an image analysis system to track a mouse under both light and dark environments. Mice were sensitized with intraperitoneal ovalbumin (OVA) injections and challenged ten times with oral OVA administration. The OVA challenges induced weight loss and diarrhea. We assessed their behavior and found that the OVA challenges decreased their total moving distance during the dark period. We also revealed that the OVA challenges increased the inactive time of mice during the dark period. Interestingly, these changes were not observed or very small during the light period. We next assessed the location of mice in the home-cage and found that the OVA challenges increased the time when mice stayed at corners and decreased the time at the center during the dark period. These observations suggest mental abnormality of mice. Indeed, the OVA challenges increased the immobility time of mice in the tail suspension test. Thus, food allergic mice exhibited reduced activity and might exhibit psychological symptoms during dark period.

Topics: Allergens; Animals; Diarrhea; Disease Models, Animal; Food Hypersensitivity; Mice; Mice, Inbred BALB C; Ovalbumin

Recent work highlighted the importance of environmental contaminants in the development of allergic diseases.. The intestinal mucosal barrier, Th (helper T) cells, DCs (dendritic cells), and intestinal flora were analyzed with flow cytometry, RNA-seq, and 16s sequencing in the present study to demonstrate whether the exposure of enterotoxins like. We found that co-exposure to SEB and Ovalbumin (OVA) could impair the intestinal barrier, imbalance the intestinal Th immune, and cause the decline of intestinal flora diversity in OVA-sensitized mice. Moreover, with the co-stimulation of SEB, the transport of OVA was enhanced in the Caco-2 cell monolayer, the uptake and presentation of OVA were promoted in the bone marrow dendritic cells (BMDCs), and Th cell differentiation was also enhanced. In summary, co-exposure to SEB in allergens should be considered a food allergy risk factor.

Topics: Animals; Caco-2 Cells; Enterotoxins; Food Hypersensitivity; Humans; Mice; Ovalbumin; Staphylococcus aureus; T-Lymphocytes, Helper-Inducer

Gut microbiota plays an important role in the regulation of food allergy. However, the interactions between the gut flora and immune system are not well studied. Here, we obtained ovalbumin (OVA)-sensitive BALB/c mice, combined with serum untargeted metabolomics to investigate the mechanisms of the interactions. The serum metabolomics results showed that 17 serum metabolites were downregulated, enriched in the aminoacyl-tRNA biosynthesis pathway, whereas indole-3-propionic acid (IPA) was increased. Six operational taxonomic units (OTUs) at the family level were altered and correlated with immune endpoints. Combined metabolomic and microbiomic analyses revealed that IPA levels were correlated with differential bacterial OTUs and a positive correlation with Treg in splenic lymphocytes. These results suggest that the regulatory effects of intestinal flora on allergic responses may be achieved by metabolizing tryptophan to produce indole derivatives and the aminoacyl-tRNA biosynthesis pathway. The formation of OVA tolerance in mice may be related to the enrichment of

Topics: Animals; Food Hypersensitivity; Gastrointestinal Microbiome; Metabolomics; Mice; Mice, Inbred BALB C; Ovalbumin

Food allergy (FA) is a common immune disorder caused by food antigens. Probiotic strains showed alleviating effects on FA, such as the alleviation of FA pathological symptoms, serum OVA-sIgE levels, and the gut microbiota diversity and composition. The results showed that intragastric administration of

Topics: Animals; Food Hypersensitivity; Gastrointestinal Microbiome; Indoles; Mice; Ovalbumin; Probiotics

Food allergy is a worldwide food safety problem with increasing prevalence. Developing novel approaches for food allergy investigations is the basis for controlling food allergies. In this work, a 3-dimensional (3D) intestinal cell model was established to simulate the intestinal mucosal immune system. Gut epithelial cell line CMT93 was cultured in a transwell insert above dendritic cells (DCs) isolated from mouse spleen and stimulated by egg allergen ovalbumin (OVA), then the conditioned media of DCs was transferred to T cells isolated from mouse spleen. The allergy-related indexes of each cell type were determined by qPCR and flow cytometry. Then the TAZ gene was knocked down in the CMT93 cells and the role of the Hippo pathway in OVA-induced food allergy was investigated. The 3D intestinal cell model showed more significant and more specific allergic responses than conventional cell models and is more convenient to be manipulated than the mouse models. This model is an ideal tool for food allergy investigations and would facilitate studies in the field of intestinal mucosal immunity.

Topics: Allergens; Animals; Food Hypersensitivity; Intestinal Mucosa; Intestines; Mice; Mice, Inbred BALB C; Ovalbumin; T-Lymphocytes

House dust mites (HDM) are one of the important factors of airway allergic diseases, HDM allergens can be detected in the human gut mucosa, which induces local inflammation and increases intestinal epithelial permeability. This study tests a hypothesis that HDM contribute to the development of OVA (ovalbumin)-induced intestinal allergy. The serum levels of IgE against HDM in patients with food allergy were detected with UniCAP100 (Pharmacia, Uppsala, Sweden); a mouse model of food allergy was developed with OVA and HDM as the specific antigens. Compared to healthy controls, patients with food allergy have higher levels of serum HDM-specific IgE. Compared to food allergy alone groups, the levels of HDM-specific IgE in patients with food allergy and asthma or allergic rhinitis were significantly higher. In mouse models, we found that HDM/OVA induced allergy-like symptoms, lower body temperature, and lower body weight. The levels of IgE, IgG1, mMCP-1 (mouse mast cell protease-1), IL-4 and IL-5 in the HDM and HDM + CT (cholera toxin) groups were higher than the control groups, and the levels of IgE, IgG1, IL-4 and IL-5 in the HDM, OVA and HDM + OVA groups were higher than the control groups. The pathological changes of intestinal tissues in the HDM and HDM + CT/the HDM, OVA and HDM + OVA groups were more severe, more eosinophil infiltration than the control groups. Moreover, exposure to HDM induced intestinal barrier dysfunction, and facilitated the development of intestinal allergy in mice. In conclusion, HDM exposure enhances immune responses to OVA-induced food allergy.

Topics: Allergens; Animals; Antigens, Dermatophagoides; Dermatophagoides pteronyssinus; Dust; Food Hypersensitivity; Humans; Immunity; Immunoglobulin E; Immunoglobulin G; Interleukin-4; Interleukin-5; Mice; Ovalbumin; Pyroglyphidae

(1) Background: Posttranslational protein modifications have been demonstrated to change protein allergenicity. Previously, it was reported that pretreatment with highly nitrated food proteins induced a tolerogenic immune response in an experimental mouse model and in human immune cells. Here, we investigated a possible therapeutic effect of modified proteins and evaluated the safety of oral exposure to highly nitrated proteins in an experimental food allergy model. (2) Methods: BALB/c mice were orally sensitized towards ovalbumin (OVA) under gastric acid suppression. Thereafter, treatment via intragastric gavage with maximally nitrated OVA (nOVAmax) and OVA as a control was performed six times every 2 weeks. On the last day of experiments, all the treated mice were orally challenged with OVA. Systemic anaphylactic reaction was determined by measuring the core body temperature. Moreover, antibody levels, regulatory T cell numbers, cytokine levels and histology of antrum tissues were analyzed. (3) Results: After oral immunotherapy, OVA-specific IgE titers were decreased while IgG1 titers were significantly elevated in the mice receiving OVA. After oral challenge with OVA, nOVAmax-treated allergic animals showed no drop of the core body temperature, which was observed for OVA-allergic and OVA-treated allergic animals. Significantly fewer eosinophils and mast cells were found in the gastric mucosa of the allergic mice after nOVAmax treatment. (4) Conclusions: Oral immunotherapy with nOVAmax reduced allergic reactions upon allergen exposure and the number of allergen effector cells in the gastric mucosa. Thus, maximally nitrated allergens enabled an efficient and safe treatment for food allergy in our experimental model.

Topics: Allergens; Animals; Disease Models, Animal; Food Hypersensitivity; Immunologic Factors; Immunotherapy; Mice; Mice, Inbred BALB C; Ovalbumin

The prevalence of food allergies has increased in Asian countries. The aim of this study was to determine the potential value of sea cucumber peptide (SCP) for anti-allergic therapeutics in terms of their effect on immune response and gut microbiota composition. Results exhibited that SCP could significantly improve the allergy symptoms caused by ovalbumin and could reduce the risk of IgE mediated allergic disorders, as well as repair the morphological damage in the colon. Flow cytometry analysis indicated that SCP could improve the ratio of CD4+/CD8+ T lymphocytes. 16S rRNA results indicated that SCP could differently impact the composition of microbiota. The relative abundances of Bacteroidetes and Firmicutes and the Bacteroidetes/Firmicutes ratio were altered in normal mice. When compared with the OVA treated group, the SCP treated groups showed an increase in the relative abundance of Lachnospiraceae, Muribaculaceae and Ruminococcaceae, and a decrease in Bacteroidaceae, Prevotellaceae, and Lactobacillaceae. These results demonstrate that SCP exhibits potential antiallergic activities in a mouse model of ovalbumin allergy by regulating intestinal microbiota diversity and upregulating the immune response of T lymphocyte subpopulations, which might provide important evidence that SCP can be developed into a novel functional food for inhibiting ovalbumin allergy.

Topics: Animals; Anti-Allergic Agents; Food Hypersensitivity; Gastrointestinal Microbiome; Immunity; Mice; Ovalbumin; RNA, Ribosomal, 16S; Sea Cucumbers

Food allergy has been one of the main problems threatening people's health in recent years. However, there is still no way to completely cure it at present. Therefore, the development of food allergy related drugs is still necessary.

Topics: Alginates; Allergens; Animals; Food Hypersensitivity; Gastrointestinal Microbiome; Humans; Immunoglobulin E; Mice; Ovalbumin; Polysaccharides; Sargassum

Epicutaneous immunotherapy (EPIT) protocols have recently been developed to restore tolerance in patients with food allergy. The mechanisms by which EPIT protocols promote desensitization rely on a profound immune deviation of pathogenic T- and B-cell responses.. To date, little is known about the contribution of skin dendritic cells (skDCs) to T-cell remodeling and EPIT efficacy.. We capitalized on a preclinical model of food allergy to ovalbumin (OVA) to characterize the phenotype and functions of OVA. Our results showed that both Langerhans cells and dermal conventional cDC1 and cDC2 subsets retained their ability to capture OVA in the skin and to migrate toward the skin-draining lymph nodes during EPIT. However, their activation/maturation status was significantly impaired, as evidenced by the gradual and selective reduction of CD86, CD40, and OVA protein expression in respective subsets. Phenotypic changes during EPIT were also characterized by a progressive diversification of single-cell gene signatures within each DC subset. Interestingly, we observed that OVA. Our results therefore emphasize that the acquisition of distinct phenotypic and functional specializations by skDCs during EPIT is at the cornerstone of the desensitization process.

Topics: Allergens; Desensitization, Immunologic; Food Hypersensitivity; Humans; Langerhans Cells; Ovalbumin; T-Lymphocytes, Regulatory

Increasing evidence indicates that intestinal microecological imbalances are strongly associated with food allergen intolerance. This study investigated the effect of olive oil on food allergy susceptibility and intestinal microecology based on an ovalbumin (OVA)-sensitized mouse model. The results indicated that the allergic symptoms of sensitized mice were alleviated when they were supplemented with olive oil at 1-3 g/kg per day for 7 weeks. Intestinal epithelium observation showed repaired ileum villi and upregulated tight junction (TJ) protein expression. Furthermore, the levels of the cytokines (e.g., IL-10) secreted by regulatory T cells were increased, whereas T helper 2 (Th2) cell-associated factors were decreased in lamina propria. Furthermore, 16S rRNA sequencing indicated reduced Burkholderiaceae and increased Clostridiaceae in the intestinal microflora. The results suggest that an olive oil-enriched diet may effectively prevent food allergies by regulating the intestinal microecological balance. PRACTICAL APPLICATIONS: Recent studies emphasized that intestinal ecological imbalance, including intestinal immunity and microflora structure, plays an important role in affecting the occurrence and development of food allergy. The present results implied that olive oil, one of the main components of the Mediterranean diet, can effectively ameliorate the symptoms of OVA-induced food allergy by regulating intestinal microecological homeostasis. Therefore, dietary supplementation with olive oil may be an effective strategy for preventing food allergies.

Topics: Allergens; Animals; Cytokines; Diet; Dietary Fats, Unsaturated; Food Hypersensitivity; Homeostasis; Interleukin-10; Mice; Mice, Inbred BALB C; Olea; Olive Oil; Ovalbumin; RNA, Ribosomal, 16S

Ovalbumin (OVA) is a food allergen whose allergenicity is modulated by heating. We aimed to establish a molecular connection between heat-induced structural modifications and the modulation of the IgE binding capacity of OVA. For this, we used model samples of heat-modified OVA with increasing complexity; glycated, aggregated, or glycated and aggregated. Using sera from egg-allergic individuals, we show that both aggregation and glycation strongly impacted IgE binding capacity, despite limited structural changes for glycated OVA. A molecular exploration at the amino acid level using high-resolution mass spectrometry revealed extensive cross-linking, mostly through disulfide and dehydroprotein bridges, and moderate but significant glycation. Structural modifications affected residues located within or at a few amino acids distance of known human linear IgE epitopes, such as C121, K123, S169, K190, K207, H332 and C368. We thus unveil key amino residues implicated in the changes in IgE binding of OVA induced by heating.

Topics: Allergens; Food Hypersensitivity; Heating; Humans; Immunoglobulin E; Mass Spectrometry; Ovalbumin

This study aimed to determine whether fucoxanthin alleviated ovalbumin (OVA)-induced food allergy (FA) and explored the possible mechanisms. The results indicated that supplementation with fucoxanthin at 10.0-20.0 mg/kg per day for 7 weeks inhibited food anaphylaxis and the production of immunoglobulin (Ig) E, IgG, histamine, and related cytokines while alleviating allergic symptoms in sensitized mice. Fucoxanthin enhanced the intestinal epithelial barrier by up-regulating tight junction (TJ) protein expression and promoting regenerating islet-derived protein III-gamma (RegIIIγ) and secretory IgA (sIgA) secretion. In addition, fucoxanthin induced the secretion of anti-inflammatory factors (interleukin (IL)-10 and transforming growth factor β (TGF-β)) by regulatory T (Treg) cells and decreased the pro-inflammatory factor levels (IL-4, tumor necrosis factor-α (TNF-α), IL-17, and IL-1β), ameliorating intestinal inflammation. Compared with the model group, beneficial bacteria, such as Lactobacillaceae, increased in the intestinal flora, while pathogenic bacteria like Helicobacteraceae, Desulfovibrionaceae, and Streptococcaceae decreased. Therefore, fucoxanthin may effectively prevent FA by enhancing the intestinal epithelial barrier and reshaping the intestinal flora.

Topics: Animals; Cytokines; Disease Models, Animal; Food Hypersensitivity; Gastrointestinal Microbiome; Immunoglobulin E; Intestinal Mucosa; Mice; Mice, Inbred BALB C; Ovalbumin; Xanthophylls

Food allergies are adverse health effects that arise from specific immune responses, occurring upon exposure to given foods, even if present in traces. Egg allergy is one of the most common food allergies, mainly caused by egg white proteins, with ovalbumin being the most abundant. As allergens can also be present in foodstuff due to unintended contamination, there is a need for analytical tools that are able to rapidly detect allergens in food products at the point-of-use. Herein, we report an origami paper-based device for detecting ovalbumin in food samples, based on a competitive immunoassay with chemiluminescence detection. In this biosensor, magnetic microbeads have been employed for easy and efficient immobilization of ovalbumin on paper. Immobilized ovalbumin competes with the ovalbumin present in the sample for a limited amount of enzyme-labelled anti-ovalbumin antibody. By exploiting the origami approach, a multistep analytical procedure could be performed using reagents preloaded on paper layers, thus providing a ready-to-use immunosensing platform. The assay provided a limit of detection (LOD) of about 1 ng mL

Topics: Allergens; Biosensing Techniques; Egg Proteins; Food Hypersensitivity; Humans; Immunoassay; Luminescence; Microspheres; Ovalbumin

This study assesses whether oleuropein prevents ovalbumin (OVA)-induced food allergy (FA) and investigates the underlying mechanisms.. A Balb/c FA mouse model is established and maintained for 7 weeks. The subjects are administered OVA by oral gavage to induce FA and supplemented with different oleuropein doses (1.00-20.00 mg kg. These findings suggest that oleuropein prevents FA by enhancing intestinal epithelial barrier function and improving immune homeostasis and intestinal flora in sensitized mice. Therefore, diets rich in oleuropein should be recommended for people with FA.

Topics: Animals; Disease Models, Animal; Food Hypersensitivity; Gastrointestinal Microbiome; Immunoglobulin E; Intestinal Mucosa; Mice; Mice, Inbred BALB C; Ovalbumin

β-Carotene is a dietary source of vitamin A, and its physiological functions, such as anti-inflammatory activity, immune regulation, and improvement of intestinal flora, are attracting increasing attention. Recent studies have shown that the development of food allergy is closely related to intestinal dysfunction. Therefore, the present study investigated the potential anti-food allergy activity of β-carotene and its regulatory intestinal homeostasis pathway. The results obtained using an ovalbumin (OVA)-induced food allergy mouse model indicated that the clinical allergic symptoms were alleviated, and the levels of anaphylactic mediators (such as immunoglobulin (Ig) E, IgG, and histamine) were reduced after β-carotene supplementation at 5.00 mg per kg per day. In addition, the expression of tight junction (TJ) proteins (claudin-1, occludin, and ZO-1) increased by 38.58%, 24.39%, and 26.23%, respectively. Additionally, the secretion of secretory IgA (sIgA) and the regeneration of islet-derived protein (Reg) IIIγ were promoted in the intestinal mucous after β-carotene administration. Furthermore, the alpha and beta diversity analysis showed that the composition and diversity of the intestinal flora in the β-carotene group tended to be normalized compared to the model group. Higher levels of beneficial bacteria, such as

Topics: Animals; beta Carotene; Food Hypersensitivity; Gastrointestinal Microbiome; Immunoglobulin E; Intestinal Mucosa; Mice; Ovalbumin; Tight Junction Proteins

A food allergy is caused by an abnormal immune reaction and can induce serious intestinal inflammation and tissue damage. Currently, the avoidance of food allergens is still the most effective way to prevent or reduce allergic symptoms, so the development of new strategies to treat allergies is important. Avenanthramide (AVA) is a bioactive polyphenol derived from oats with a wide range of biological activities; however, it is still not clear whether or how AVA alleviates intestinal damage under allergic situations. The aim of this study was to explore the effect of AVA on the small intestinal damage in an ovalbumin (OVA)-induced food allergy model and its mechanism. In experiment 1, 10 mg/kg bw and 20 mg/kg bw doses of AVA both decreased the serum levels of OVA-specific IgE, histamine, and prostaglandin D induced by OVA. The AVA administration relieved inflammation indicated by the lower serum concentrations of pro-inflammatory cytokines including interleukin-1β, IL-6, and tumor necrosis factor-α. The levels of tight junction proteins including Claudin-1, ZO-1, and Occludin in the jejunum were elevated after AVA administration, accompanied by the improved intestinal morphology. Furthermore, AVA elevated the protein expression of heat shock protein 70 (Hsp70) and inhibited the phosphorylation of nuclear factor kappa-B (NF-κB), thus the apoptozole, which a Hsp70 inhibitor, was applied in experiment 2 to assess the contribution of Hsp70-NF-κB signaling to the effects of AVA. In the experiment 2, the inhibition of Hsp70 signaling treatment abolished the beneficial effects of AVA on the small intestinal damage and other allergic symptoms in mice challenged with OVA. Taken together, our results indicated that AVA exerted an intestinal protection role in the OVA-induced allergy, the mechanism of which was partly mediated by the Hsp70-NF-κB signaling.

Topics: Animals; Cytokines; Food Hypersensitivity; HSP70 Heat-Shock Proteins; Intestine, Small; Mice; NF-kappa B; Ovalbumin; Signal Transduction

The significant increase in food allergy incidence is correlated with dietary changes in modernized countries. Here, we investigated the impact of dietary emulsifiers on food allergy by employing an experimental murine model. Mice were exposed to drinking water containing 1.0% carboxymethylcellulose (CMC) or Polysorbate-80 (P80) for 12 weeks, a treatment that was previously demonstrated to induce significant alterations in microbiota composition and function leading to chronic intestinal inflammation and metabolic abnormalities. Subsequently, the ovalbumin food allergy model was applied and characterized. As a result, we observed that dietary emulsifiers, especially P80, significantly exacerbated food allergy symptoms, with increased OVA-specific IgE induction and accelerated type 2 cytokine expressions, such as IL-4, IL-5, and IL-13, in the colon. Administration of an antibiotic regimen completely reversed the emulsifier-induced exacerbated susceptibility to food allergy, suggesting a critical role played by the intestinal microbiota in food allergy and type 2 immune responses.

Topics: Animals; Colon; Diet; Disease Models, Animal; Emulsifying Agents; Food Hypersensitivity; Immunity; Inflammation; Mice; Mice, Inbred BALB C; Ovalbumin; Polysorbates

The global demand of sustainable food sources leads to introduction of novel foods on the market, which may pose a risk of inducing allergic sensitization. Currently there are no validated. This. Ovalbumin activates IEC and underlying moDCs, both resulting in downstream IgE isotype-switching. However, only direct exposure of moDCs to ovalbumin drives Th2 polarization and a humoral B cell response allowing for IgE mediated mast cell degranulation, IL13 and IL4 release in this sequential DC-T cell-B cell-mast cell model, indicating also an immunomodulatory role for IEC.

Topics: Allergens; Food Hypersensitivity; Humans; Immunoglobulin E; Intestines; Ovalbumin

Topics: Animals; Anti-Allergic Agents; Antibodies, Monoclonal; Disease Models, Animal; Food Hypersensitivity; Histamine Release; Mice, Inbred BALB C; Ovalbumin; Peptides; Rabbits; Tumor Protein, Translationally-Controlled 1

Delivery by cesarean section (CS) is linked to an increased incidence of food allergies in children and affects early gut microbiota colonization. Furthermore, emerging evidence has connected disordered intestinal microbiota to food allergies. Here, we investigated the impact of CS on a rat model for food allergy to ovalbumin (OVA). Rats delivered by CS were found to be more responsive to OVA sensitization than vaginally born ones, displaying a greater reduction in rectal temperature upon challenge, worse diarrhea, and higher levels of OVA-specific antibodies and histamine. 16S rRNA sequencing of feces revealed reduced levels of

Topics: Allergens; Animals; Bifidobacterium; Cells, Cultured; Cesarean Section; Disease Models, Animal; Dysbiosis; Female; Food Hypersensitivity; Gastrointestinal Microbiome; Humans; Immunoglobulin E; Lactobacillus; Male; Ovalbumin; Pregnancy; Probiotics; Rats; Rats, Sprague-Dawley; RNA, Ribosomal, 16S; Th2 Cells; Tight Junctions

(1) Background: The use of antibiotics affects the composition of gut microbiota. Studies have suggested that the colonization of gut microbiota in early life is related to later food allergies. Still, the relationship between altered intestinal microbiota in adulthood and food allergies is unclear. (2) Methods: We established three mouse models to analyze gut microbiota dysbiosis' impact on the intestinal barrier and determine whether this effect can increase the susceptibility to and severity of food allergy in later life. (3) Results: The antibiotic-induced gut microbiota dysbiosis significantly reduced Lachnospiraceae, Muribaculaceae, and Ruminococcaceae, and increased Enterococcaceae and Clostridiales. At the same time, the metabolic abundance was changed, including decreased short-chain fatty acids and tryptophan, as well as enhanced purine. This change is related to food allergies. After gut microbiota dysbiosis, we sensitized the mice. The content of specific IgE and IgG1 in mice serum was significantly increased, and the inflammatory response was enhanced. The dysbiosis of gut microbiota caused the sensitized mice to have more severe allergic symptoms, ruptured intestinal villi, and a decrease in tight junction proteins (TJs) when re-exposed to the allergen. (4) Conclusions: Antibiotic-induced gut microbiota dysbiosis increases the susceptibility and severity of food allergies. This event may be due to the increased intestinal permeability caused by decreased intestinal tight junction proteins and the increased inflammatory response.

Topics: Animals; Anti-Bacterial Agents; Biodiversity; Disease Models, Animal; Disease Susceptibility; Dysbiosis; Female; Food Hypersensitivity; Gastrointestinal Microbiome; Haptoglobins; Inflammation; Injections, Intraperitoneal; Intestines; Metabolome; Mice; Mice, Inbred BALB C; NF-kappa B; Ovalbumin; Phylogeny; Protein Precursors; Receptor, PAR-2; Severity of Illness Index; Tight Junction Proteins

Mounting evidence demonstrates that a high-salt diet (HSD) not only affects hemodynamic changes but also disrupts immune homeostasis. The T helper 17 (Th17) and regulatory T cells (Tregs) are susceptible to hypersalinity. However, research on the influence of sodium on Th2-mediated food allergies remains scarce. We aimed to investigate the effect of dietary sodium on the immune response to food allergies. Mice maintained on an HSD (4% NaCl), low-salt diet (LSD; 0.4% NaCl), or control diet (CTRL; 1.0% NaCl) were orally sensitized with ovalbumin (OVA) and a cholera toxin (CT) adjuvant, and then subjected to an intragastric OVA challenge. OVA-specific immunoglobulin G (IgG), IgG1, IgG2a, and IgE antibodies were significantly higher in the HSD group than in the CTRL group (

Topics: Animal Nutritional Physiological Phenomena; Animals; Diet; Diet, Sodium-Restricted; Disease Models, Animal; Food Hypersensitivity; Immunity; Mice; Ovalbumin; Salt Tolerance; Sodium, Dietary; T-Lymphocytes, Regulatory; Th2 Cells

Deoxynivalenol (DON), a highly prevalent contaminant of grain-based products, is known to induce reproductive- and immunotoxicities. Considering the importance of immune development in early life, the present study investigated the effects of perinatal DON exposure on allergy development and vaccine responsiveness in the offspring. Pregnant mice received control or DON-contaminated diets (12.5 mg/kg diet) during pregnancy and lactation. After weaning, female offspring were sensitized to ovalbumin (OVA) by oral administration of OVA with cholera toxin (CT). Male offspring were injected with Influvac vaccine. OVA-specific acute allergic skin response (ASR) in females and vaccine-specific delayed-type hypersensitivity (DTH) in males were measured upon intradermal antigen challenge. Immune cell populations in spleen and antigen-specific plasma immunoglobulins were analyzed. In female CT+OVA-sensitized offspring of DON-exposed mothers ASR and OVA-specific plasma immunoglobulins were significantly higher, compared to the female offspring of control mothers. In vaccinated male offspring of DON-exposed mothers DTH and vaccine-specific antibody levels were significantly lower, compared to the male offspring of control mothers. In both models a significant reduction in regulatory T cells, Tbet

Topics: Animals; Antibodies, Viral; Cells, Cultured; Cholera Toxin; Cytokines; Disease Models, Animal; Female; Food Hypersensitivity; Immunogenicity, Vaccine; Influenza Vaccines; Lactation; Male; Maternal Exposure; Mice, Inbred C3H; Ovalbumin; Pregnancy; Prenatal Exposure Delayed Effects; Spleen; T-Lymphocytes, Regulatory; Th1 Cells; Trichothecenes; Vaccination; Vaccine Efficacy

Oral immunotherapy (OIT) aims to establish desensitization and sustained unresponsiveness (SU) in patients with food allergy by ingestion of gradually increasing doses of specific food allergens. However, little is known about the mechanisms by which OIT induces SU to specific allergens.. We investigated the role of Notch signaling, which controls cell fate decisions in many types of immune cells in the induction of SU by OIT treatment.. Two types of mouse models, ovalbumin-induced food allergy and OIT, were generated. To elucidate the role of Notch signaling in OIT-induced SU, mice were intraperitoneally injected with the Notch signaling inhibitor N-[(3,5-difluorophenyl)acetyl]-l-alanyl-2-phenylglycine-1,1-dimethylethyl ester during the OIT treatment period.. Ovalbumin-sensitized mice were desensitized and also had SU induced by OIT treatment, whereas repeated challenges with ovalbumin caused the development of severe allergic reactions in ovalbumin-sensitized mice. Administration of N-[(3,5-difluorophenyl)acetyl]-l-alanyl-2-phenylglycine-1,1-dimethylethyl ester to mice during the OIT treatment period inhibited the establishment of SU to ovalbumin but did not affect the induction of desensitization. OIT induced a systemic expansion of IL-10-producing CD4. Notch signaling contributes to the establishment of SU induced by OIT through systemic expansion of immunosuppressive cells, such as IL-10-producing CD4

Topics: Administration, Oral; Allergens; Animals; Cells, Cultured; Desensitization, Immunologic; Disease Models, Animal; Female; Food Hypersensitivity; Humans; Immune Tolerance; Interleukin-10; Mice; Mice, Inbred BALB C; Myeloid-Derived Suppressor Cells; Ovalbumin; Receptors, Notch; Signal Transduction; Th2 Cells

Food allergy is an antigen-specific immunological adverse reaction after exposure to a given food. Multiple clinical studies showed that oral immunotherapy (OIT) is effective for the prevention and treatment for food allergy that is developed in infants and children. However, the effectiveness of OIT for epicutaneously sensitized food allergy remains unclear. Previously, we established a mouse model of epicutaneous-sensitized food allergy. In this model, systemic allergic reaction including intestinal and skin symptoms, such as anaphylaxis, was observed. We treated this model with OIT in two ways (OIT before sensitization or OIT during the sensitization phase) and evaluated the preventive effect of both methods. OIT before sensitization significantly ameliorated mast cell degranulation in sensitized skin, but there was no decrease in rectal temperatures or in mast cell degranulation in the jejunum. However, OIT administered during the sensitization phase significantly ameliorated the decrease in rectal temperature and mast cell degranulation in the skin and jejunum. OIT before sensitization increased the regulatory T cells in mesenteric lymph node (MLN), but not in the spleen, and it reduced antigen-specific IgG, but not IgE, production compared with the non-OIT control. However, OIT during sensitization caused a greater increase in regulatory T cells in both the MLN and spleen and reduced antigen-specific IgE and IgG generation compared with the non-OIT control group. Thus, OIT during the sensitization phase was effective for the prevention of epicutaneous-sensitized food allergy.

Topics: Administration, Cutaneous; Administration, Oral; Anaphylaxis; Animals; Antigens; Body Temperature; Cell Degranulation; Chymases; Desensitization, Immunologic; Disease Models, Animal; Food Hypersensitivity; Immune Tolerance; Immunoglobulin E; Immunoglobulin G; Jejunum; Lymph Nodes; Mast Cells; Mesentery; Mice; Ovalbumin; Skin; Skin Diseases; Spleen; T-Lymphocytes, Regulatory

Hippo signaling is a crucial pathway in innate immune responses, but the relationship between food allergy and Hippo pathway is unknown. The aim of this work is to investigate the regulation of food allergy by Hippo pathway and reveal the molecular mechanisms.. Two food allergens tropomyosin and ovalbumin are used to challenge a mouse model and CMT93 intestinal epithelia cell model. The allergic responses and the activation of Hippo pathway are tested in these models. In the mouse model, both allergens trigged significant allergic responses, and Hippo pathway is suppressed after allergen challenge. In CMT93, both allergens upregulate the expression of allergic cytokines thymic stromal lymphopoietin, interleukin (IL)-25, and IL-33. In TAZ KD CMT93, the Hippo pathway is blocked, and the expression of allergenic cytokines are also suppressed.. Both in vivo and in vitro data demonstrate that the two food allergens suppressed Hippo pathway by downregulating TAZ expression, resulting in intestinal epithelia instability, and finally leading to hypersensitivity reactions. These findings provide potential therapeutic targets and molecular markers for food allergy, and provide dietary guidelines for allergenic individuals.

Topics: Adaptor Proteins, Signal Transducing; Animals; Cell Line; Cysteine-Rich Protein 61; Cytokines; Female; Food Hypersensitivity; Hippo Signaling Pathway; Intestinal Mucosa; Mice, Inbred BALB C; Ovalbumin; Protein Serine-Threonine Kinases; Thymic Stromal Lymphopoietin; Tropomyosin

Polyunsaturated fatty acids (PUFAs) are present in biological membranes and influence membrane fluidity and immune responses. PUFAs such as 18:2n-6 and 18:3n-3 cannot be synthesized de novo in mammals and are thus called essential fatty acids (EFAs). In addition, PUFAs can be converted to very long-chain PUFAs (VLC-PUFAs), such as arachidonic acid and docosahexaenoic acid, in the body. Although avoiding allergens is an effective strategy for food-allergy patients, the dietary exclusion of several allergens reportedly induces deficiencies in essential nutrients such as PUFAs. In this study, we investigated whether an EFA-deficient (EFAD) diet influenced allergic symptoms in ovalbumin (OVA)-immunized mice. Unexpectedly, no exacerbation of immune responses after OVA-sensitization was observed in mice fed an EFAD diet, and no differences in serum PUFA levels between OVA-immunized and non-immunized mice fed the EFAD diet were detected. However, levels of VLC-PUFAs in the small intestine increased after OVA-sensitization and did not decrease during EFAD diet administration, showing that small intestinal VLC-PUFAs levels were strongly preserved in the food-allergy model mice. Further studies are required to elucidate the mechanisms by which small intestinal VLC-PUFAs are retained in food-allergy model mice.

Topics: Animals; Disease Models, Animal; Fatty Acids, Essential; Food Hypersensitivity; Immunization; Intestine, Small; Male; Mice; Mice, Inbred BALB C; Ovalbumin

Up to 20% of people worldwide develop gastrointestinal symptoms following a meal

Topics: Abdominal Pain; Adult; Allergens; Animals; Citrobacter rodentium; Diarrhea; Enterobacteriaceae Infections; Female; Food; Food Hypersensitivity; Glutens; Humans; Immunoglobulin E; Intestinal Mucosa; Intestines; Irritable Bowel Syndrome; Male; Mast Cells; Mice; Mice, Inbred BALB C; Middle Aged; Milk; Ovalbumin; Quality of Life; Receptors, Histamine H1; Soybean Proteins; Triticum

The role of serum S100A8/A9 in intestinal inflammation has been confirmed, and its role in food allergy is currently being investigated.. To explore the levels of S100A8/A9 and inflammatory factors, including Toll-like receptors 4 (TLR4), Nuclear transcription factors (NF-κB) and Tumor necrosis factor α (TNF-α), in mild food allergies.. Eighty 3-week-old male Brown Norway rats were used. Forty rats were randomly assigned to the ovalbumin-sensitized experimental group, while 40 rats were assigned to the normal saline sham-sensitized control group. Body weight and length and the levels of serum ovalbumin-specific IgE (OVA-IgE), histamine, Th1-associated and Th2-associated factors, S100A8/A9 and inflammation-associated cytokines were compared.. Through the evaluation of OVA-IgE level and Th1/Th2 balance in the experimental group, a successful IgE-mediated food allergy model was constructed. Compared with the control group, the experimental group had higher serum S100A8/A9 levels on days 21, 28, 35 and 42 (all P < 0.05); higher TLR4 levels on days 28, 35 and 42 (all P < 0.05); higher TNF-α levels on days 28, 35 and 42 (all P < 0.05); higher NF-κB levels on days 35 and 42 (all P < 0.05); and higher IL-1β and IL-6 levels on days 7 to 42 (all P < 0.05). Moreover, positive correlations were found between the serum levels of S100A8/A9 and inflammation-associated cytokines [TNF-α: r = 0.378, P = 0.039; IL-1β: r = 0.679, P = 0.000; IL-6: r = 0.590, P = 0.001].. S100A8/A9 and inflammatory-related factors, including TLR4, NF-κB, TNF-α, IL-6 and IL-1β, is closely related to food allergies. Moreover, immune and inflammatory factors interact with each other in food allergies, which may provide insight into food allergy causes and treatments.

Topics: Animals; Calgranulin A; Calgranulin B; Cytokines; Food Hypersensitivity; Immunoglobulin E; Inflammation; Inflammation Mediators; Male; NF-kappa B; Ovalbumin; Rats; Rats, Inbred BN; Th1-Th2 Balance; Toll-Like Receptor 4; Tumor Necrosis Factor-alpha

Topics: Animals; Anti-Allergic Agents; Anti-Inflammatory Agents; Antioxidants; Cytokines; Female; Food Hypersensitivity; Immunoglobulin E; Mice; Mice, Inbred BALB C; Ovalbumin; Piper nigrum; Plant Extracts; T-Lymphocytes, Regulatory; Th17 Cells; Th2 Cells

Impaired gastric digestion due to suppressed gastric acidity enhances the risk for food allergy development. In the current study, we aimed to evaluate the impact of a supported gastric digestion via application of a pharmaceutical gastric enzyme solution (GES) on food allergy development and allergic reactions in a BALB/c mouse model. The ability of the GES to restore hypoacidic conditions was tested in mice treated with gastric acid suppression medication. To evaluate the impact on allergic symptoms, mice were orally sensitized with ovalbumin (OVA) under gastric acid suppression and subjected to oral challenges with or without GES. The immune response was evaluated by measurement of antibody titers, cytokine levels, mucosal allergy effector cell influx and regulatory T-cell counts. Clinical response was objectified by core body temperature measurements after oral OVA challenge. Supplementation of GES transiently restored physiological pH levels in the stomach after pharmaceutical gastric acid suppression. During oral sensitization, supplementation of gastric enzymes significantly reduced systemic IgE, IgG1 and IgG2a levels and allergic symptoms. In food allergic mice, clinical symptoms were reduced by co-administration of the gastric enzyme solution. Support of gastric digestion efficiently prevents food allergy induction and alleviates clinical symptoms in our food allergy model.

Topics: Allergens; Animals; Antibodies; B-Lymphocyte Subsets; Cytokines; Dietary Supplements; Digestion; Disease Models, Animal; Food Hypersensitivity; Gastrointestinal Agents; Immune Tolerance; Lymphocyte Count; Mice; Mice, Inbred BALB C; Ovalbumin; Stomach; T-Lymphocytes, Regulatory

Fructooligosaccharides (FOS) can change gut microbiota composition and play a protective role in food allergy (FA). Furthermore, the protective mechanism of FOS against FA is unclear. In this study, intestinal flora and tryptophan (Trp) metabolites were investigated in a mouse model with FA supplemented with FOS. Meanwhile, we injected aryl hydrocarbon receptor antagonists (AhR-A) into a mouse model of FA supplemented with FOS to investigate whether T helper cell (Th) 17/regulatory T (Treg) cell balance was affected. Our research studies showed that dietary intake of FOS provided moderate protection from the intestinal inflammation induced by ovalbumin (OVA). This protective effect disappeared in AhR-A mice. The OVA mice manifestations had significantly lower bacterial richness, when compared to the normal control (NC) mice. Among fecal bacteria, the abundance of Akkermansiaceae (family level) and Verrucomicrobia (phylum level) increased and Ruminococcacere (phylum level) decreased in the feces of allergic mice. These changes were reversed by FOS treatment. FOS modulated the gut microbiome profiles that were altered in OVA mice, which showed an increase in the abundance of Ruminococcacere (phylum level) and a decrease in the abundance of Akkermansiaceae (family level) and Verrucomicrobia (phylum level). Liquid chromatography/tandem mass spectrometry (LC-MS/MS) analysis of Trp metabolites showed significant reductions in the level of kynurenine (kyn) in the serum of OVA mice, as compared to NC and FOS mice. Conversely, the levels of Trp and 5-hydroxytryptamine (5-HT) were significantly increased in OVA mice. Correlation analysis revealed a negative relationship between the relative abundance of Verrucomicrobiae (class level) and Akkermansiaceae (family level) with kyn, and a positive relationship with 5-HT. FOS significantly reduced interleukin-17A (IL-17A) and retinoic acid-associated nuclear orphan receptor-γt (RORγt) in FOS mice but not in AhR-A mice. FOS increased the level of interleukin-10 (IL-10) and Forkhead box P3 (Foxp3) in FOS mice but not in AhR-A mice. These findings suggest that FOS ameliorates allergic symptoms and impacts Th17/Treg balance in mice by modulating the gut microbiota composition and Trp metabolites. FOS may serve as an effective tool for the treatment of FA by regulating immune and gut microbiota.

Topics: Animals; Disease Models, Animal; Food Hypersensitivity; Male; Mice; Mice, Inbred BALB C; Oligosaccharides; Ovalbumin; T-Lymphocytes, Regulatory; Th17 Cells; Tryptophan

Food allergies are usually managed by food avoidance. Hidden allergens in food, due to cross-contamination and/or allergenic additives added during production, place an important concern in today's increasing food allergy cases worldwide. Previous studies showed that the introduction of unacquainted food components, in an inflamed intestine, results in sensitization to this food. Thus, our aim was to evaluate the kinetics of multiple food allergy induction. Adult male C57BL/6 mice were divided into five groups, four of which were submitted to an intestinal inflammation induction protocol to peanuts. Egg white (OVA) diluted 1:5 v/v in distilled water was instilled by gavage 6h-before (PRIOR), concomitant (AT) and 6h-after (DURING) the onset of the peanut challenge diet. Positive control (POS CONT) and NEG CONT received saline per gavage. Finally, animals were challenged with subcutaneous injections of OVA. Results showed no changes in diet intake were observed. Anti-OVA polyisotypic IgG antibody titers significantly increased in AT. Flow cytometry revealed significant decrease in CD4

Topics: Allergens; Animals; Biomarkers; Cytokines; Disease Models, Animal; Disease Susceptibility; Food Hypersensitivity; Gastroenteritis; Humans; Immune Tolerance; Immunization; Immunoglobulin G; Immunomodulation; Lymphocyte Subsets; Mice; Organ Specificity; Ovalbumin

We previously described that Porphyra haitanensis sulfated polysaccharide (PHSP) maintains the balance of pro-inflammation and immunosuppression. However, it is unclear whether degraded PHSP (DPHSP) still shows the immunomodulatory activity. Here, we degraded PHSP by four different methods alone or combined in pairs, and the results showed that the molecular weight and viscosity of DPHSP were significantly decreased, while the main chemical bonds and functional structure were consistent with those of PHSP. We then investigated the immunomodulatory function of DPHSP in vitro and in vivo. Actually, DPHSP enhances the inhibitory effects on mast cell activation and improves the suppression activity of PHSP on the food anaphylactic response. In an ovalbumin-induced food allergy mouse model, the production of allergic mediators and cytokines (interleukin-4 and 13, and interferon-γ) was inhibited by DPHSP. Meanwhile, DPHSP had a stronger ability to up-regulate the differentiation of regulatory T (Treg) cells and its related cytokines. These results suggested that DPHSP showed a better anti-food allergic ability than PHSP by regulating T helper cell balance and promoting Treg cell differentiation, which indicates that DPHSP is a novel potential nutrient component against food allergy.

Topics: Anaphylaxis; Animals; Anti-Allergic Agents; Antibodies; Cell Differentiation; Cytokines; Female; Food Hypersensitivity; Histamine; Mice; Mice, Inbred BALB C; Ovalbumin; Polysaccharides; Porphyra; Sulfates; T-Lymphocytes, Helper-Inducer; T-Lymphocytes, Regulatory

Topics: Abdominal Pain; Animals; Citrobacter rodentium; Disease Models, Animal; Enterobacteriaceae Infections; Food Hypersensitivity; Humans; Immunoglobulin E; Irritable Bowel Syndrome; Mast Cells; Mice; Ovalbumin

In the previous study, pickle-derived Lactiplantibacillus plantarum 22A-3 (LP22A3) suppressed ear edema in passive cutaneous anaphylaxis by its oral administration. Moreover, LP22A3 treatment directly to RBL-2H3 cells shows no effect on β-hexosaminidase release from RBL-2H3 but inhibited its release using the Caco-2/RBL-2H3 cells co-culture system stimulated with LP22A3 from the apical side. In this study, oral administration of LP22A3 decreased total IgE and ovalbumin (OVA) specific IgE contents in blood of BALB/c mice induced food allergy by OVA. Moreover, its oral administration suppressed the development of dermatitis induced by 2,4-dinitrochlorobenzene (DNCB) which was used to develop atopic dermatitis-like lesions in NC/Nga mice. This alleviation was further correlated with a reduction of elevated serum total IgE, transepidermal water loss and elevated acanthosis in the LP22A3-treated group compared with vehicle-treated positive group. In co-culture system composed of Caco-2 and RBL-2H3 cells, LP22A3 treatment on apical side before or after the sensitization with anti-dinitrophenyl (DNP) IgE antibody indicated the different effect on β-hexosaminidase release from RBL-2H3. Its treatment before the sensitization decreased β-hexosaminidase release, but not after sensitization, indicating that LP22A3 affected mast cells sensitized with allergen through intestinal epithelial cells. These results suggest that LP22A3 may have a potential therapeutic property for Type 1 hypersensitivity and atopic dermatitis.

Topics: Animals; Caco-2 Cells; Dermatitis, Atopic; Dinitrochlorobenzene; Fermented Foods; Food Hypersensitivity; Humans; Mast Cells; Mice; Mice, Inbred BALB C; Ovalbumin

This research aimed to investigate the allergic reaction of C3H/HeJ mice after sensitization with ovalbumin (OVA) without any adjuvant and to analyze the association between intestinal microbiota and allergy-related immune cells in mesenteric lymph nodes (MLN). The allergic responses of C3H/HeJ mice orally sensitized with OVA were evaluated, and immune cell subsets in spleen and MLN and cytokines were also detected. The intestinal bacterial community structure was analyzed, followed by Spearman correlation analysis between changed gut microbiota species and allergic parameters. Sensitization induced a noticeable allergic response to the gavage of OVA without adjuvant. Increased levels of Th2, IL-4, CD103

Topics: Animals; Bacteria; Cytokines; Dendritic Cells; Disease Models, Animal; Feces; Food Hypersensitivity; Gastrointestinal Microbiome; Lymph Nodes; Mesentery; Mice; Mice, Inbred C3H; Ovalbumin; Spleen; T-Lymphocytes, Regulatory

Food allergy is associated with a high personal health and economic burden. For immunomodulation toward tolerance, food compounds could be chemically modified, for example, by posttranslational protein nitration, which also occurs via diet-derived nitrating agents in the gastrointestinal tract.. We sought to analyze the effect of pretreatment with nitrated food proteins on the immune response in a mouse food allergy model and on human monocyte-derived dendritic cells (moDCs) and PBMCs.. The model allergen ovalbumin (OVA) was nitrated in different nitration degrees, and the secondary structures of proteins were determined by circular dichroism (CD). Allergy-preventive treatment with OVA, nitrated OVA (nOVA), and maximally nitrated OVA (nOVAmax) were performed before mice were immunized with or without gastric acid-suppression medication. Antibody levels, regulatory T-cell (Treg) numbers, and cytokine levels were evaluated. Human moDCs or PBMCs were incubated with proteins and evaluated for expression of surface markers, cytokine production, and proliferation of Th2 as well as Tregs.. In contrast to OVA and nOVA, the conformation of nOVAmax was substantially changed. nOVAmax pretreated mice had decreased IgE as well as IgG1 and IgG2a levels and Treg numbers were significantly elevated, while cytokine levels remained at baseline level. nOVAmax induced a regulatory DC phenotype evidenced by a decrease of the activation marker CD86 and an increase in IL-10 production and was associated with a higher proliferation of memory Tregs.. Oral pretreatment with highly nitrated proteins induces a tolerogenic immune response in the food allergy model and in human immune cells.

Topics: Administration, Oral; Allergens; Animals; Blood Donors; Cells, Cultured; Cytokines; Dendritic Cells; Disease Models, Animal; Female; Food Hypersensitivity; Humans; Immune Tolerance; Immunization; Immunoglobulin E; Mice; Mice, Inbred BALB C; Nitro Compounds; Ovalbumin; Signal Transduction; T-Lymphocytes, Regulatory

Esophagitis with eosinophilia, inflammation, and fibrosis represent a chronic condition in humans with food allergies.. In this investigation, we asked whether esophagitis with an eosinophilic component is observed in young pigs rendered allergic to hen egg white protein (HEWP).. Food allergy was induced in young pigs using two protocols. In one protocol, sensitized pigs were challenged by gavage with a single dose of HEWP. Clinical signs were monitored for 24 hours, and then, gastrointestinal (GI) tissues were collected for histological examination. The phenotype of circulating, ovalbumin (OVA)-specific T cells also was examined in HEWP challenged animals. In the second protocol, sensitized animals were fed HEWP for 28 days. Animals were then examined by endoscopy and gastrointestinal tissues collected for histological examination.. In pigs challenged by gavage with HEWP, clinical signs were noted in 5/6 pigs including diarrhoea, emesis, and skin rash. Clinical signs were not seen in any control group. Histological analysis revealed significant levels of oesophageal eosinophilic infiltration (P < .05) in 4/6 of these animals, with two also displaying eosinophilic infiltration in the stomach. Eosinophils were not increased in ileum or colon samples. Increased numbers of circulating, OVA-specific CD4. Food allergy in the pig can be associated with esophagitis based on histological and endoscopic findings, including eosinophilic infiltration. The young pig may, therefore, be a useful large animal model for the study of eosinophilic esophagitis in humans.

Topics: Animals; CD4-Positive T-Lymphocytes; Colon; Diarrhea; Disease Models, Animal; Egg Hypersensitivity; Egg Proteins; Endoscopy, Digestive System; Eosinophilic Esophagitis; Eosinophils; Esophagus; Exanthema; Food Hypersensitivity; Ileum; Immunophenotyping; Ovalbumin; Sus scrofa; Vomiting

Inotodiol is a lanostane triterpenoid found only in Chaga mushroom. In the previous study investigating anti-allergic effects of fractionated Chaga mushroom extracts, we have found evidence that purified inotodiol holds an activity to suppress the mast cell function in vivo. To address the therapeutic relevance of the finding, in this study, we investigated whether inotodiol could also alleviate allergy symptoms observed in a chicken ovalbumin (cOVA)-induced mouse model of food allergy. Like the crude 70% ethanol extract of Chaga mushroom (320 mg/kg), oral administration of inotodiol (20 mg/kg), regardless of whether that was for preventive or treatment purpose, resulted in a significant improvement in allergic symptoms and inflammatory lesions in the small intestine appearing after repeated oral challenge with cOVA. Despite the results that inotodiol (20 mg/kg) and the Chaga mushroom extract (320 mg/kg) took effect to a similar extent, immunological mechanisms underlying those effects were found to be distinct from each other. That is, the results obtained from several in vivo assays, including mast cell-mediated passive systemic anaphylaxis, activation/proliferation of adoptively transferred antigen-specific T cells and immunoglobulin (IgG1, IgE, IgA) production by antigen-specific B cells, illustrated that inotodiol selectively inhibited the mast cell function without having any noticeable effect on other immune responses while the crude Chaga mushroom extract indiscriminately suppressed diverse immune responses. The strong anti-allergic activity of inotodiol, along with its remarkable selectivity to mast cell, makes it an excellent therapeutic candidate for food allergy with both high efficacy and outstanding safety.

Topics: Allergens; Animals; Anti-Allergic Agents; Cell Degranulation; Disease Models, Animal; Food Hypersensitivity; Humans; Inonotus; Lanosterol; Mast Cells; Mice; Mice, Inbred BALB C; Ovalbumin; Triterpenes

The therapies for food allergy (FA) need to be improved. The generation of inducible regulatory T cells (Tregs) can support immune tolerance in the body. This study aims to suppress experimental FA by inducing Tregs through the employment of modified exosomes (mExosomes). In this study, mExosomes were prepared by incubating dendritic cells with interleukin (IL)-2 and ovalbumin (OVA, used as a specific antigen) in the culture. Exosomes were purified from culture supernatant and used as the mExosomes. A murine FA model was developed to test the effects of mExosomes on the generation of Tregs in the mouse intestinal tissues and inhibiting FA. The results showed that mExosomes, which carried IL-2 and a complex of OVA peptide-major histocompatibility complex class II on the surface of exosomes, bound to OVA-specific CD4

Topics: Animals; Dendritic Cells; Exosomes; Food Hypersensitivity; Immune Tolerance; Interleukin-2; Intestines; Mice; Mice, Inbred BALB C; Ovalbumin; T-Lymphocytes, Regulatory

There has been a marked increase in life-threatening food allergy (FA). One hypothesis is that changes in bacterial communities may be key to FA. To better understand how gut microbiota regulates FA in humans, we established a mouse model with FA induced by ovalbumin. We found that the mice with FA had abnormal bacterial composition, accompanied by increased immunoglobulin G, immunoglobulin E, and interleukin-4/interferon-γ, and there existed a certain coherence between them. Interestingly, Bifidobacterium breve M-16V may alter the gut microbiota to alleviate the allergy symptoms by IL-33/ST2 signaling. Our results indicate that gut microbiota is essential for regulating FA to dietary antigens and demonstrate that intervention in bacterial community regulation may be therapeutically related to FA.

Topics: Animals; Bifidobacterium breve; Food Hypersensitivity; Gastrointestinal Microbiome; Interleukin-33; Mice, Inbred BALB C; Ovalbumin; Probiotics; Signal Transduction

It is known that the immune tolerance can be naturally established in the intestine, while the mechanism by which the immune tolerance development in the intestine is not fully understood yet. Vasoactive intestinal peptides (VIP) has the immune regulatory functions. This study aims to investigate the role of VIP in the immune tolerance development in the intestine.. Intestinal epithelial cell (IEC)-derived exosomes were prepared. The exosomes carried IL-10 and antigen/MHC II complexes. VIP-deficient (VIPd) mice and wild type mice were employed to test the role of VIP in the development of immune tolerance in the intestine.. VIPd mice failed to induce type 1 regulatory T cells (Tr1 cells) in the intestine and retarded the establishment of antigen (Ag)-specific immune tolerance. Exposure to VIP in the culture induced IL-10 expression in intestinal epithelial cells (IECs). Exosomes derived from ovalbumin (OVA, used as a specific Ag)/VIP-primed IECs carried IL-10 and OVA/MHC II complexes; these exosomes were designated IL10CARs (IL-10/chimeric antigen receptor-carrying exosomes). IL10CARs could recognize OVA-specific CD4. The data show that IL10CARs are capable of suppressing experimental FA by inducing antigen-specific Tr1 cells, which has the translation potential for FA treatment.

Topics: Animals; Antigens; CD4-Positive T-Lymphocytes; Epithelial Cells; Exosomes; Food Hypersensitivity; Histocompatibility Antigens Class II; Immune Tolerance; Interleukin-10; Intestinal Mucosa; Mice; Mice, Inbred BALB C; Ovalbumin; T-Lymphocytes, Regulatory; Vasoactive Intestinal Peptide

Allergic disorders, characterized by Th2 immune responses to environmental substances, are increasingly common in children in Western societies. Multiple studies indicate that breastfeeding, early complementary introduction of food allergens, and antibiotic avoidance in the first year of life reduces allergic outcomes in at-risk children. Why the benefit of these practices is restricted to early life is largely unknown. We identified a preweaning interval during which dietary antigens are assimilated by the colonic immune system. This interval is under maternal control via temporal changes in breast milk, coincides with an influx of naive T cells into the colon, and is followed by the development of a long-lived population of colonic peripherally derived Tregs (pTregs) that can be specific for dietary antigens encountered during this interval. Desynchronization of mothers and offspring produced durable deficits in these pTregs, impaired tolerance to dietary antigens introduced during and after this preweaning interval, and resulted in spontaneous Th2 responses. These effects could be rescued by pTregs from the periweaning colon or by Tregs generated in vitro using periweaning colonic antigen-presenting cells. These findings demonstrate that mothers and their offspring are synchronized for the development of a balanced immune system.

Topics: Allergens; Animals; Animals, Newborn; Antigen-Presenting Cells; Colon; Female; Food Hypersensitivity; Immune Tolerance; Immunoglobulin G; Mice; Mice, Inbred C57BL; Milk; Mothers; Ovalbumin; T-Lymphocytes, Regulatory; Th2 Cells; Weaning

Effectiveness of retinoic acid (RA) in treating food allergy is not yet clear. Using an allergic mouse model, we examined the amelioration of the severity of food allergy by daily RA intake with allergen or without. Female Balb/c mice were systemically sensitized to egg white (EW) and alum by intraperitoneal injection. Sensitized mice were provided diets supplemented with 0% (non-treated group), 0.1% EW (allergen group), 0.0017% RA (RA group), or 0.1% EW plus 0.0017% RA (RA+allergen group) with 20% casein for 4 wk. Oral food challenge (OFC) and allergic biomarkers were quantified. The decrease in rectal temperature post-OFC was significantly suppressed in the RA and RA+allergen groups compared to those in the non-treated and allergen groups, respectivety. The plasma levels of ovalbumin-specific IgE, IgA and IgG1 at the study endpoint were higher in the allergen and RA+allergen groups than those in the non-treated and RA+allergen groups, respectivety. Plasma ovalbumin-specific IgG2a levels at the study endpoint were significantly higher in the RA+allergen group than those in the RA groups. The supernatant concentrations of interleukin-10 and interferon-γ in the cultured spleen lymphocytes were highest in the RA+allergen group compared to those in the other groups. Thus, continuous intake of RA under allergen exposure ameliorated the severity of food allergy in a mouse model with food allergy.

Topics: Allergens; Animals; Body Temperature; Dietary Supplements; Disease Models, Animal; Egg Hypersensitivity; Egg White; Female; Food Hypersensitivity; Immunoglobulin A; Immunoglobulin E; Immunoglobulin G; Interferon-gamma; Interleukin-10; Lymphocytes; Mice; Mice, Inbred BALB C; Ovalbumin; Tretinoin

Glycomacropeptide (GMP) is a bioactive peptide derived from milk κ-casein with immune-modulatory and anti-inflammatory properties. Food allergy (FA) is an adverse immune reaction with a broad spectrum of manifestations. Allergen intake induces persistent intestinal inflammation and tissue damage. In this study, the anti-allergic activity of GMP was evaluated using a rat ovalbumin (OVA)-induced FA model with gastrointestinal manifestation. Rats were orally GMP treated from 3 days prior and during FA development. The severity of food anaphylaxis and diarrheal episodes, antibody production and histamine level were measured. Histopathological changes, inflammation and predominant cytokine profile at intestine were analyzed. Oral GMP intake decreased clinical signs and diarrhea severity induced by allergen, with a significant reduction in intestinal edema and expression level of

Topics: Allergens; Anaphylaxis; Animals; Anti-Allergic Agents; Caseins; Cytokines; Disease Models, Animal; Down-Regulation; Food Hypersensitivity; GATA3 Transcription Factor; Interleukin-13; Interleukin-1beta; Interleukin-5; Intestines; Male; Mast Cells; Ovalbumin; Peptide Fragments; Rats; Rats, Wistar

Viridicatol is a quinoline alkaloid isolated from the deep-sea-derived fungus

Topics: Anaphylaxis; Animals; Anti-Allergic Agents; Aquatic Organisms; B-Lymphocytes; beta-N-Acetylhexosaminidases; Calcium; Cell Line, Tumor; Disease Models, Animal; Food Hypersensitivity; Histamine; Hydroxyquinolines; Immunoglobulin E; Interleukin-10; Intestines; Mast Cells; Mice; Ovalbumin; Penicillium; Peptide Hydrolases; Quinolones; Rats; T-Lymphocytes, Regulatory; Tumor Necrosis Factor-alpha

Nevadensin (NEV), a natural flavonoid compound derived from Lysionotus pauciflorus Maxim, has numerous biological activities. However, few researchers have examined its potential impact on alleviating allergies. In the present study, NEV was found to upregulate rectal temperature, suppress the development of diarrhea, and decrease the levels of serum specific immunoglobulin E, histamine and mouse MC protease-1 in ovalbumin-allergic mice. Moreover, NEV also alleviated passive cutaneous anaphylaxis reactions and inhibited the release of β-hexosaminidase and histamine in bone marrow-derived mast cells. Furthermore, we provide the first demonstration that NEV decreases the expression of c-Kit and suppresses the proliferation of bone marrow-derived mast cells and accelerates their apoptosis. These findings indicated that L. pauciflorus-derived NEV might have the potential to alleviate food hypersensitivity.

Topics: Animals; beta-N-Acetylhexosaminidases; Cell Line; Cell Proliferation; Cell Survival; Cytokines; Disease Models, Animal; Flavones; Food Hypersensitivity; Histamine; Immunoglobulin E; Mice; Mice, Inbred BALB C; Ovalbumin; Passive Cutaneous Anaphylaxis; Proto-Oncogene Proteins c-kit

Isoflavones have many biological activities and are major bioactive components of kakkonto, a traditional Japanese herbal medicine. We previously reported that the combined therapy of oral immune therapy (OIT) and kakkonto downregulates the mRNA expression of Cyp26b1, a major retinoic acid (RA)-degrading enzyme, in the colon of food allergy mice and thereby ameliorates allergic symptoms. In this study, we evaluated the effects of various isoflavones on Cyp26b1 expression in primary cultured lamina propria (LP) cells isolated from the mouse colon. The mRNA expression of Cyp26b1 was extremely downregulated by all isoflavones tested in the LP cells except for puerarin. In particular, genistein and genistin markedly suppressed Cyp26b1 mRNA expression without affecting RA-synthesizing enzyme expression. Moreover, to evaluate the effects of isoflavones on allergic reactions, genistein and genistin were administered to ovalbumin (OVA)-induced food allergy mice. Oral administration of genistin suppressed the development of allergic symptoms. These results raise the possibility that isoflavones elevated the level of RA in the colon by inhibiting RA degradation and then the high concentration of RA in the colon might exert immunosuppressive and antiallergic effects on food allergy mice.

Topics: Animals; Colon; Food Hypersensitivity; Gene Expression Regulation, Enzymologic; Intestinal Mucosa; Isoflavones; Mice; Mice, Inbred BALB C; Mucous Membrane; Ovalbumin; Retinoic Acid 4-Hydroxylase

Silkworm droppings have long been used in traditional medicine to remedy allergic itching, palsy, blood circulation problems, and arthritis in Asian countries. To investigate the anti-allergic effect of silkworm dropping extract (SDE) and its mechanism, we used a mouse model of food allergy induced by ovalbumin (OVA).. SDE ameliorated the symptoms of OVA-induced food allergies, and the levels of T helper 2 (Th2)-related cytokines [such as interleukin (IL)-4, IL-5, IL-10, and IL-13] were found to be significantly decreased in both the spleen and mesenteric lymph nodes by SDE. Furthermore, SDE treatment directly inhibited OVA permeation, IL-4 production, and degranulation of mast cells; in contrast, immunoglobulin E (IgE) production from B cells was not affected.. These results suggest that SDE has potential anti-allergic activities, and SDE may be useful in the treatment/prevention of allergic disorders such as food allergies, serving as therapeutic agents. © 2019 Society of Chemical Industry.

Topics: Animals; Anti-Allergic Agents; Bombyx; Feces; Female; Food Hypersensitivity; Humans; Immunoglobulin E; Interleukin-13; Interleukin-4; Interleukin-5; Mast Cells; Mice; Mice, Inbred BALB C; Ovalbumin; Th2 Cells

The prevalence of IgE-mediated food allergy is increasing in the whole wide world which often causes skin and gastrointestinal tract symptoms, or even fatal anaphylactic shock. However, the evaluation of food allergens remains difficult, and the mechanism of food allergy is still not fully clear. To study the gene expression profile in food allergy animal models and identify the regulatory mechanism of the crucial genes, two administration routes were used to build animal models in our study. OVA-specific IgE and IL-4 levels were tested by ELISA, transcriptome profiling was carried out by microarray, and the regulatory mechanism of the highest expressed gene was studied in the primary spleen cells. We found that activation-induced cytidine deaminase (Aicda) is the highest expressed gene in the allergic mice, IL-21 can dramatically enhance the expression of Aicda in the lymph node microenvironment, and IL-17A can promote this effect significantly though it has only limited influence by itself. At last, we illuminated that the promotion of IL-21 on Aicda is partially through STAT3. In summary, our results suggest that IL-21 and IL-17A may play important role in the expression of Aicda as well as food allergy.

Topics: Allergens; Anaphylaxis; Animals; Cytidine Deaminase; Cytokines; Disease Models, Animal; Female; Food Hypersensitivity; Immunoglobulin E; Interleukins; Male; Mice; Mice, Inbred BALB C; Mice, Inbred C3H; Ovalbumin; Spleen

Topics: Animals; Anti-Allergic Agents; Antrodia; Benzodioxoles; Cells, Cultured; Cytokines; Dendritic Cells; Disease Models, Animal; Food Hypersensitivity; Hypersensitivity; Mice, Inbred BALB C; Ovalbumin; Phytotherapy; Th2 Cells

Food allergies have become a major healthcare concern, hence preventive efforts to ensure oral tolerance induction to newly introduced antigens are particularly relevant. Given that transforming growth factor-β (TGF-β) plays a key role in immune tolerance, we tested whether an infant formula enriched with TGF-β would improve oral tolerance induction. A partially hydrolyzed whey protein-based formula was enriched with cow's-milk-derived TGF-β (TGF-β-enriched formula) by adding a specific whey protein isolate (WPI). The manufacturing process was optimized to achieve a concentration of TGF-β within the range of human breast milk concentrations. Protection from allergic sensitization and immune response was assessed in a mouse model. Adult mice received the TGF-β-enriched formula, a control non-enriched formula, or water ad libitum for 13 days before sensitization and suboptimal tolerization to ovalbumin (OVA). When compared to non-tolerized mice, suboptimally-tolerized mice supplemented with the TGF-β-enriched formula showed significantly lower levels of total immunoglobulin-E (IgE) and OVA-specific (IgG1). Mouse mast-cell protease-1 (mMCP-1) and cytokine levels were also significantly decreased in suboptimally-tolerized mice fed the TGF-β-enriched formula. In conclusion, oral supplementation with cow's-milk-derived TGF-β decreased allergic responses to newly introduced allergens and thus reduced the risk of developing food allergy.

Topics: Allergens; Animals; Cattle; Chymases; Cytokines; Female; Food Hypersensitivity; Humans; Immune Tolerance; Immunoglobulin G; Infant; Infant Formula; Mice; Mice, Inbred C57BL; Milk; Ovalbumin; Transforming Growth Factor beta; Whey Proteins

Coumarin is an important organic heterocyclic compound with a wide range of sources in nature. It plays an important role in the drug discovery process due to its existence in diverse biologically active compounds and its broad bioactivity. In this study, the anti-allergic activity of coumarin was evaluated using an ovalbumin (OVA)-induced mouse food allergy model and an immunoglobulin (Ig)E mediated mouse bone marrow-derived mast cell (BMMC) model. Coumarin could alleviate the OVA-induced allergic symptoms, decrease the diarrhea rates, and promote the rectal temperature rise in allergic mice. Moreover, coumarin had the ability to reduce the levels of histamine and mouse mast cell proteinases, inhibit OVA-specific IgE, and significantly decrease the population of mast cells in the spleen and mesenteric lymph nodes. Coumarin could also significantly suppress mast cell-dependent passive cutaneous anaphylaxis. Additionally, the number of mature BMMCs was decreased as coumarin caused the suppression of c-KIT receptors. Furthermore, coumarin up-regulated the apoptosis of OVA-activated BMMCs in a concentration-dependent manner. In conclusion, coumarin displayed effective anti-food allergy activity via the regulation of mast cell function and numbers. Coumarin and its derivatives provide a new direction for the development of anti-food allergic drug components.

Topics: Anaphylaxis; Animals; Anti-Allergic Agents; Cell Line; Coumarins; Disease Models, Animal; Female; Food Hypersensitivity; Histamine; Immunoglobulin E; Mast Cells; Mice; Mice, Inbred BALB C; Ovalbumin; Passive Cutaneous Anaphylaxis; Spleen

Dihydromyricetin (DMY) is a natural flavonoid compound derived from Lysionotus pauciflorus Maxim and has been found to possess numerous biological activities. However, there have been few reports regarding its anti-food allergic activity. In this study, we demonstrated that DMY could upregulate the rectal temperature, suppress the development of diarrhea, decrease the levels of serum specific immunoglobulin (Ig)E, histamine, and mouse mast cell protease-1, and promote the production of interleukin-10 in ovalbumin-allergic mice. Moreover, DMY downregulated the population of B cells and mast cells and upregulated the population of regulatory T cells in the spleens of ovalbumin-allergic mice. Furthermore, DMY blocked the high affinity IgE receptor (FcεRI)-IgE interaction, inhibited the release of β-hexosaminidase and histamine in rat basophilic leukemia-2H3 cells, and alleviated passive cutaneous anaphylaxis reactions. These findings indicated that L. pauciflorus derived DMY might have the potential to alleviate food hypersensitivity or allergic diseases.

Topics: Animals; Anti-Allergic Agents; B-Lymphocytes; Female; Flavonols; Food Hypersensitivity; Humans; Immunoglobulin E; Mast Cells; Mice; Mice, Inbred BALB C; Ovalbumin; Passive Cutaneous Anaphylaxis

Tibetan medicine has been practiced for 3800 years. Anzhijinhua San (AZJHS), which is a traditional Tibetan medicine, has been effective in the treatment of indigestion, anorexia and cold diarrhea. However, the effects of AZJHS on allergic diarrhea have not been reported.. The aim of the present study was to elucidate the effect of AZJHS on experimental ovalbumin-induced diarrhea and elucidate its possible mechanism.. Female BALB/c mice were sensitized by intraperitoneal injection with 50 μg ovalbumin (OVA) and 1 mg alum in saline twice during a 2-week period. From day 28, mice were orally challenged with OVA (50 mg) every other day for a total of ten times. AZJHS (46.8 and 468.0 mg/kg) was orally administered every other day from day 0-46. Food allergy symptoms were evaluated. OVA- specific IgE, 5-HT and its metabolites in serum were determined. Immunohistochemical and histopathology were performed in gastrointestinal tract tissues. 5-HT-related gene expression was assayed in the colon.. Severe symptoms of allergic diarrhea were observed in the model group (diarrhea, anaphylactic response, and rectal temperature). AZJHS (46.8 and 468.0 mg/kg) significantly reduced mouse diarrhea and significantly prevented the increases in OVA-specific IgE levels (P < 0.05), which challenge with OVA. AZJHS (46.8 and 468.0 mg/kg) significantly prevented the increases in 5-HT-positive cells. The nuclei of EC cells in the AZJHS (46.8 and 468.0 mg/kg) group increased in size and the secretory granules were fewer in number compared with those in the model group. AZJHS (46.8 and 468.0 mg/kg) significantly increased the relative fold changes of 5-HTP and 5-HT compared with the model group. The mRNA expression of the serotonin transporter (Sert) and serotonin receptor 3A (Htr3a) was significantly decreased after the 10th challenge with OVA, and AZJHS (46.8 and 468.0 mg/kg) significantly increased these levels.. We demonstrated that the administration of AZJHS attenuated OVA-induced diarrhea by regulating the serotonin pathway. These results indicated that AZJHS may be a potential candidate as an anti-allergic diarrhea agent.

Topics: Animals; Anti-Allergic Agents; Diarrhea; Disease Models, Animal; Female; Food Hypersensitivity; Humans; Medicine, Tibetan Traditional; Mice; Mice, Inbred BALB C; Ovalbumin; Plant Extracts; Serotonin; Signal Transduction; Treatment Outcome

Can f 1 belongs to the lipocalin superfamily and is considered to be an animal allergen. The immune response induced by Can f 1 in mice was compared with that induced by ovalbumin (OVA), a typical food allergen. Female BALB/c and C57BL/6 mice (6 weeks of age) were subcutaneously injected with Can f 1 or OVA with or without aluminum hydroxide (Alum) three times with intervals of two weeks. Serum levels of total IgE or antigen-specific IgE and production of IL13 and IFNγ from splenocytes were analyzed. Immunization with Can f 1 or OVA increased serum levels of both total IgE and antigen-specific IgE significantly irrespective of Alum. These results indicate that Can f 1 and OVA were able to induce allergic sensitization in mice. Splenocyte production of IL13 in mice immunized with Can f 1 or OVA with and without Alum were significantly increased after stimulation with each antigen. However, IL13 levels in the mice immunized with Can f 1 with Alum were significantly lower than those immunized without Alum. Increases in IFNγ levels after stimulation with Can f 1 or OVA were not remarkable. No influence of genetic backgrounds of BALB/c and C57BL/6 mice was found. Although Can f 1 induced Th2 type immune responses as was also the case for immunization with OVA, an inhibitory effect of Alum on induction of IL13 was observed only in mice immunized with Can f 1. These results suggest that the immune mechanism for allergic sensitization with Can f 1 is different from that with OVA.

Topics: Allergens; Animals; Antibody Specificity; Cytokines; Disease Models, Animal; Female; Food Hypersensitivity; Immunization; Immunoglobulin E; Mice; Mice, Inbred BALB C; Mice, Inbred C57BL; Ovalbumin; Species Specificity; Spleen

The vagus nerve has emerged as an important modulator of the intestinal immune system. Its anti-inflammatory properties have been previously shown in innate and Th1/Th17 predominant inflammatory models. To what extent the vagus nerve is of importance in Th2 inflammatory responses like food allergy is still unclear. In this study, we therefore aimed to investigate the effect of vagotomy (VGX) and vagus nerve stimulation (VNS), on the development and severity of experimental food allergy.. Balb/C mice were first sensitized with ovalbumin (OVA) in the presence of alum. Prior to oral challenges with OVA, mice were subjected to VGX or VNS. Disease severity was determined by assessing severity and onset of diarrhoea, OVA-specific antibody production, mast cell number and activity, inflammatory gene expression in duodenal tissue and lamina propria immune cells by flow cytometry analysis.. When compared to control mice, VGX did not significantly affect the development and severity of the disease in our model of food allergy. VNS, on the other hand, resulted in a significant amelioration of the different inflammatory parameters assessed. This effect was independent of α7nAChR and is possibly mediated through the dampening of mast cells and increased phagocytosis of OVA by CX3CR1. These results underscore the anti-inflammatory properties of the vagus nerve and the potential of neuro-immune interactions in the intestine. Further insight into the underlying mechanisms could ultimately lead to novel therapeutic approaches in the treatment of not only food allergy but also other immune-mediated diseases.

Topics: Allergens; alpha7 Nicotinic Acetylcholine Receptor; Animals; Biomarkers; Cell Membrane Permeability; Disease Models, Animal; Food Hypersensitivity; Gastroenteritis; Immunophenotyping; Macrophages; Mast Cells; Mastocytosis; Mice; Mice, Knockout; Neutrophil Infiltration; Ovalbumin; Severity of Illness Index; Vagotomy; Vagus Nerve Stimulation

Topics: Albumins; Amaranthus; Animals; Antibodies, Anti-Idiotypic; Chymases; Flour; Food Handling; Food Hypersensitivity; Globulins; Immunoglobulin E; Immunoglobulin G; Mice; Mice, Inbred BALB C; Models, Animal; Ovalbumin

Resveratrol exists widely in plant species and has a variety of anti-oxidant, anti-inflammatory, and immunomodulatory properties. However, there have been few reports regarding its anti-food allergic activity. In this study, we demonstrated that resveratrol (isolated from Abies georgei) could decrease the release of β-hexosaminidase and histamine in rat basophilic leukemia-2H3 cells. Resveratrol was not only found to suppress the development of diarrhea, up-regulate the rectal temperature of ovalbumin-allergic mice, and decrease the serum level of specific immunoglobulin E, mouse mast cell protease-1 and histamine, but also found to decrease the population of dendritic cells, B cells and mast cells of ovalbumin -allergic mice in the spleen or mesenteric lymph node. Furthermore, resveratrol inhibited the release of β-hexosaminidase and histamine in bone marrow-derived cells and alleviated mast cell-mediated passive cutaneous anaphylaxis reactions. These findings indicated that resveratrol isolated from Abies georgei might have the potential to alleviate food hypersensitivity or allergic disease.

Topics: Abies; Animals; Anti-Allergic Agents; B-Lymphocytes; beta-N-Acetylhexosaminidases; Cell Line; Disease Models, Animal; Female; Food Hypersensitivity; Histamine; Humans; Immunoglobulin E; Mast Cells; Mice; Mice, Inbred BALB C; Ovalbumin; Passive Cutaneous Anaphylaxis; Peptide Hydrolases; Plant Extracts; Rats; Resveratrol

The role of dysbiosis in food allergy (FA) remains unclear. We found that dysbiotic fecal microbiota in FA infants evolved compositionally over time and failed to protect against FA in mice. Infants and mice with FA had decreased IgA and increased IgE binding to fecal bacteria, indicative of a broader breakdown of oral tolerance than hitherto appreciated. Therapy with Clostridiales species impacted by dysbiosis, either as a consortium or as monotherapy with Subdoligranulum variabile, suppressed FA in mice as did a separate immunomodulatory Bacteroidales consortium. Bacteriotherapy induced expression by regulatory T (Treg) cells of the transcription factor ROR-γt in a MyD88-dependent manner, which was deficient in FA infants and mice and ineffectively induced by their microbiota. Deletion of Myd88 or Rorc in Treg cells abrogated protection by bacteriotherapy. Thus, commensals activate a MyD88/ROR-γt pathway in nascent Treg cells to protect against FA, while dysbiosis impairs this regulatory response to promote disease.

Topics: Animals; Bacteroides; Clostridiales; Dysbiosis; Feces; Food Hypersensitivity; Gastrointestinal Microbiome; Mice; Mice, Inbred BALB C; Mice, Inbred C57BL; Myeloid Differentiation Factor 88; Nuclear Receptor Subfamily 1, Group F, Member 3; Ovalbumin; Signal Transduction; T-Lymphocytes, Regulatory

Raw cow's milk was previously shown to suppress allergic symptoms in a murine model for food allergy. In the present study, we investigated the contribution of fat content and heat-sensitive milk components to this allergy-protective effect. In addition, we determined the potency of alkaline phosphatase (ALP), a heat-sensitive raw milk component, to affect the allergic response. C3H/HeOuJ mice were treated with raw milk, pasteurized milk, skimmed raw milk, pasteurized milk spiked with ALP, or phosphate-buffered saline for eight days prior to sensitization and challenge with ovalbumin (OVA). Effects of these milk types on the allergic response were subsequently assessed. Similar to raw milk, skimmed raw milk suppressed food allergic symptoms, demonstrated by a reduced acute allergic skin response and low levels of OVA-specific IgE and Th2-related cytokines. This protective effect was accompanied by an induction of CD103

Topics: Alkaline Phosphatase; Animals; Basophils; Cells, Cultured; Cytokines; Dermatitis, Atopic; Disease Models, Animal; Female; Food Handling; Food Hypersensitivity; Immunoglobulins; Lipids; Lymph Nodes; Mice, Inbred C3H; Milk Proteins; Ovalbumin; Pasteurization; Protein Denaturation; Skin; Spleen

Epidemiological studies identified raw cow's milk consumption as an important environmental exposure that prevents allergic diseases. In the present study, we investigated whether raw cow's milk has the capacity to induce tolerance to an unrelated, non-milk, food allergen. Histone acetylation of T cell genes was investigated to assess potential epigenetic regulation. Female C3H/HeOuJ mice were sensitized and challenged to ovalbumin. Prior to sensitization, the mice were treated with raw milk, processed milk, or phosphate-buffered saline for eight days. Allergic symptoms were assessed after challenge and histone modifications in T cell-related genes of splenocyte-derived CD4

Topics: Acetylation; Animals; Cells, Cultured; Chromatin Assembly and Disassembly; Cytokines; Disease Models, Animal; Epigenesis, Genetic; Female; Food Hypersensitivity; Histones; Immune Tolerance; Lymphocyte Activation; Mice, Inbred C3H; Milk; Ovalbumin; T-Lymphocyte Subsets; T-Lymphocytes, Regulatory; Th1 Cells; Th2 Cells

Topics: Allergens; Animals; Diarrhea; Disease Models, Animal; Female; Food Hypersensitivity; Immunoglobulin E; Immunoglobulin G; Interleukin-4; Lymph Nodes; Mice, Inbred BALB C; Ovalbumin; Vitamin D Deficiency

As documented in a Vietnamese traditional medical encyclopedia, Syzygium formosum (Wall.) Masam leaves have been routinely used among indigenous Vietnamese people for treatment of various allergy-like symptoms including dermatitis and rhinitis.. Anti-allergic activity of S. formosum leaves was examined with a mouse model of chicken ovalbumin (cOVA)-induced food allergy, and mechanisms underlying the anti-allergic effect were explored.. BALB/c mice were administered i.p. cOVA (20μg) plus alum (2mg) twice on day 0 and 14 for sensitization (immunization). Two weeks after the second immunization, the mice were administered cOVA (50mg) p.o. 5 times every 3 days to induce food allergy symptoms (i.e., anaphylaxis, diarrhea, and drop in the body temperature). Ethanol extract of dried leaves of S. formosum (80mg/kg or 200mg/kg body weight) was administered p.o. daily during the induction (challenge) period.. Treatment with the S. formosum leaves ethanol extract ameliorated the allergic symptoms to a significant extent and in a dose-dependent manner. The treatment also resulted in a significant improvement in the inflammatory lesion in the small intestine and reduction in the numbers of mast cells and eosinophils recruited to the lesion. The treatment also brought about a significant reduction in the levels of Th2 cytokines produced by the mesenteric lymph node cells cultured ex vivo with cOVA. The passive anaphylaxis experiment also showed that the extract treatment impaired the mast cell function.. Our study provides a scientific basis for the traditional (indigenous) use of the S. formosum leaves extract for the treatment of various allergy symptoms in Vietnam. In addition, the results show that the extract has activities to suppress antigen-specific Th2 T cell immune responses and the mast cell function, which are directly related with its anti-allergic effect.

Topics: Allergens; Alum Compounds; Animals; Anti-Allergic Agents; Chymases; Cytokines; Ethanol; Female; Flavonoids; Food Hypersensitivity; Immunoglobulin E; Immunoglobulin G; Lymph Nodes; Mice, Inbred BALB C; Ovalbumin; Phytotherapy; Plant Extracts; Plant Leaves; Solvents; Syzygium; Triterpenes

Atopic dermatitis (AD) is recently increasing among populations, but the underlying mechanisms remain controversial. Interactions between the gut microbiota and mucosal immunity are considered to be a crucial etiology. Fructooligosaccharide (FOS), prebiotics have been reported as activators of the gut microbiota.. The aim of this study was to investigate the effects of kestose, the smallest FOS and FOS on atopic dermatitis in mice.. An AD mouse model was developed by (ovalbumin) epidermal sensitization using BALB/c mice. Kestose (1%, 5%, and 10%) or FOS (5%, positive control) was orally administered throughout the study.. In comparison with the values observed for the control AD mice, transepidermal water loss (TEWL), clinical score, and skin inflammation on histopathology were significantly decreased by the oral administration of kestose. Total IgE, thymic stromal lymphopoietin (TSLP) in skin, and IL-4 were also suppressed by this administration. In addition, the population of CD4. These findings suggest that kestose activates the gut immune system to induce the tolerance against allergic skin inflammations in AD.

Topics: Animals; CD4-Positive T-Lymphocytes; Dermatitis, Atopic; Disease Models, Animal; Epidermis; Female; Food Hypersensitivity; Gastrointestinal Microbiome; Humans; Immune Tolerance; Immunity, Mucosal; Intestinal Mucosa; Mice; Mice, Inbred BALB C; Ovalbumin; Trisaccharides

The role of maternal immune responses in tolerance induction is poorly understood. To study whether maternal allergen sensitization affects offspring susceptibility to food allergy, we epicutaneously sensitized female mice with ovalbumin (OVA) followed by epicutaneous sensitization and oral challenge of their offspring with OVA. Maternal OVA sensitization prevented food anaphylaxis, OVA-specific IgE production, and intestinal mast cell expansion in offspring. This protection was mediated by neonatal crystallizable fragment receptor (FcRn)-dependent transfer of maternal IgG and OVA immune complexes (IgG-IC) via breast milk and induction of allergen-specific regulatory T (T reg) cells in offspring. Breastfeeding by OVA-sensitized mothers or maternal supplementation with IgG-IC was sufficient to induce neonatal tolerance. FcRn-dependent antigen presentation by CD11c

Topics: Allergens; Animals; Antigen-Antibody Complex; Dendritic Cells; Disease Models, Animal; Female; Food Hypersensitivity; Histocompatibility Antigens Class I; Immune Tolerance; Immunization; Immunoglobulin G; Maternal Exposure; Mice; Mice, Knockout; Ovalbumin; Pregnancy; Prenatal Exposure Delayed Effects; Receptors, Fc; T-Lymphocytes, Regulatory

While an anti-allergic effect of Chaga mushroom (Inonotus obliquus) has been indicated, its therapeutic effect on allergy and immunoregulatory mechanisms and chemical constituents directly responsible for that are hardly known. We examined the effect of 70% ethanol extract of Chaga mushroom (EE) and its dichloromethane (DF) and aqueous (AF) fractions using a mouse model of chicken ovalbumin (cOVA)-induced food allergy, and found that only EE and DF ameliorated allergy symptoms to a significant extent. The in vivo mast cell-stabilizing activity was also found only in EE and DF whereas the activities to suppress Th2 and Th17 immune responses and cOVA-specific IgE production in the small intestine were observed in all three treatment regimens, implying that inhibition of the mast cell function by lipophilic compounds was vital for the therapeutic effect. Results also indicated that inotodiol, a triterpenoid predominantly present in DF, played an active role as a mast cell stabilizer.

Topics: Animals; Anti-Allergic Agents; Basidiomycota; Disease Models, Animal; Ethanol; Female; Food Hypersensitivity; Humans; Immunoglobulin E; Lanosterol; Mast Cells; Methylene Chloride; Mice; Mice, Inbred BALB C; Ovalbumin; Th17 Cells; Th2 Cells

Interleukin (IL)-15 overexpression in eosinophilic gastrointestinal disorders is reported, but IL-15's role in promoting eosinophilic gastroenteritis is largely unknown. Therefore, we generated enterocyte-overexpressed IL-15 transgenic mice using Fabpi promoter. The Fabpi-IL-15 (iIL-15) transgenic mice showed induced IL-15 levels in the jejunum with a marked increase in jejunum eosinophils. However, no induction of eosinophilia in the blood or any other gastrointestinal segment was observed. Eosinophilia in the jejunum villus was substantially higher in iIL-15 mice compared to wild-type mice. In addition, goblet cell hyperplasia was also observed in the jejunum of iIL-15 mice. Furthermore, a significant correlation between induced IL-15 transcript and the IL-18 transcripts was observed. Therefore, to further understand the role of IL-18 in IL-15 mice associated gastrointestinal disorders, we generated iIL-15/IL-18Rα

Topics: Allergens; Animals; Colon; Cytokines; Eosinophilia; Esophagus; Food Hypersensitivity; Goblet Cells; Hyperplasia; Immunoglobulins; Interleukin-15; Interleukin-18; Intestines; Mice, Inbred BALB C; Mice, Transgenic; Organ Specificity; Ovalbumin; Promoter Regions, Genetic; Rats; Th2 Cells

Food allergy is a major public health problem. Studies have shown that long-term interactions between activated leucocyte cell adhesion molecule (ALCAM/CD166) on the surface of antigen-presenting cells, and CD6, a co-stimulatory molecule, influence immune responses. However, there are currently no studies on the functions of ALCAM in food allergy. Therefore, we aimed to identify the functions of ALCAM in ovalbumin (OVA)-induced food allergy using ALCAM-deficient mice. Wild-type (WT) and ALCAM-deficient (ALCAM

Topics: Activated-Leukocyte Cell Adhesion Molecule; Adoptive Transfer; Animals; Antigens, CD; Antigens, Differentiation, T-Lymphocyte; Cell Proliferation; Child; Child, Preschool; Coculture Techniques; Cytokines; Dendritic Cells; Disease Models, Animal; Female; Food Hypersensitivity; Gene Expression Regulation; Humans; Immunoglobulin E; Lymphocyte Activation; Male; Mice; Mice, Inbred BALB C; Mice, Knockout; Ovalbumin; Th2 Cells

Direct contact of food proteins with eczematous lesions is thought to be the main cause of epicutaneous sensitization. To further investigate the development and pathogenesis of food allergy in vivo, a good mouse model of epicutaneous sensitization is needed. However, a fundamental problem in that regard is that the optimal age for epicutaneous sensitization of mice is unknown. In this study, we attempted to elucidate that optimal age.. Dorsal skin of wild-type BALB/c female mice (1, 3, 8 and 24 weeks old) was shaved, depilated and tape-stripped. A Finn chamber containing a 20-μl-aliquot of 20-mg/ml (OVA) was applied to the tape-stripped skin on 3 consecutive days/week, for 3 weeks. The body temperature was measured after intraperitoneal OVA challenge. Serum OVA-specific IgE titers and OVA-induced cytokine production by spleen cells were measured by ELISA. Dendritic cells (DCs) that migrated to the draining lymph nodes were quantified by FITC-labeled OVA and flow cytometry. The mRNA expression levels in the dorsal skin were measured by qPCR.. A significant age-dependent body temperature decline was observed after OVA challenge. The serum OVA-specific IgE titer, OVA-induced cytokine production (i.e., IL-4, IL-5 and IL-13) by spleen cells, and number of FITC-OVA-engulfing DCs increased with age. In addition, mRNA for IL-33, but not TSLP or IL-25, was significantly induced in the skin by tape-stripping and increased with age.. Twenty-four-week-old mice showed the greatest DC migration, Th2 polarization, IgE production and body temperature decline. Skin-derived IL-33 is likely to play key roles in those changes.

Topics: Administration, Cutaneous; Animals; Disease Models, Animal; Female; Food Hypersensitivity; Immunization; Mice; Mice, Inbred BALB C; Ovalbumin; Skin

An allergic reaction occurs when the immune system overreacts to harmless substance called allergen that gains access to the body. Food allergy is a hypersensitive immune reaction to food proteins and the number of patients with food allergy has recently increased. Aloe Vera is used for wellness and medicinal purposes. In particular, Aloe vera has been reported to enhance immunity. However, the effect of Aloe vera on food allergy is not yet known. In this study, we investigated the effects of processed Aloe vera gel (PAG) containing low molecular weight Aloe polysaccharide (AP) on ovalbumin (OVA)-induced food allergy in mice. Allergic symptoms, rectal temperature, and diarrhea were measured in OVA-induced food allergy mice. Other allergic parameters were also analyzed by RT-PCR, ELISA, flow cytometry, and other biochemical methods. As the results, PAG suppressed the decrease of body temperature, diarrhea, and allergic symptoms in OVA-induced food allergy mice. PAG also reduced serum concentrations of type 2 helper T cell (Th2) cytokines (Interleukin-(IL)-4, IL-5, and IL-13) as well as histamine, mast cell protease-1 (MCP-1), and immunoglobulin (Ig)E. PAG blocked the degranulation of mast cells and infiltration of eosinophils in intestine. Furthermore, PAG suppressed the population of Th2 cells in spleen and mesenteric lymph nodes. PAG also increased the production of IL-10 and population of type 1 regulatory T (Tr1) cells in mice with food allergy. Taken together, our findings suggest that PAG suppressed Th2 immune responses through, at least partially, stimulating the secretion of IL-10 in food allergy mice.

Topics: Animals; Cytokines; Disease Models, Animal; Female; Food Hypersensitivity; Immunoglobulin E; Intestines; Mast Cells; Mice; Mice, Inbred BALB C; Ovalbumin; Plant Preparations; Polysaccharides; Spleen; Th2 Cells

Most of the patients develop food allergy early in life. The factors related to parental immune condition might be one of the conceivable causes.. We reported murine models of food allergy and oral OVA tolerance. To investigate the influence of parental immune condition on infant food allergy, female and male mice with food allergy or oral tolerance were mated with each other.. Food allergy was suppressed by decreased IgE production in the offspring of mice with food allergy. On the contrary, anaphylaxis for OVA was induced in the offspring of mice with oral tolerance. The suppression of food allergy being dependent on a maternal factor was revealed in the offspring after cross-mating mice with food allergy and oral tolerance. Because OVA-specific IgG, presumed to be from the allergic mother, was detected in the serum of naïve infants from mothers allergic to food, we assumed that the suppression was dependent on a specific IgG. The serum IgG purified by a G-protein column was administered before OVA sensitization in the food allergy model, and OVA-specific IgE production was found to be diminished in the administered mice. However, OVA-specific monoclonal IgG. We demonstrated that maternal specific IgG conjugated food antigen is an important factor related to the development of food allergy and acquiring tolerance.

Topics: Allergens; Anaphylaxis; Animals; Antigen-Antibody Complex; Disease Models, Animal; Female; Food Hypersensitivity; Immune Tolerance; Immunoglobulin A; Immunoglobulin E; Immunoglobulin G; Male; Mice, Inbred BALB C; Ovalbumin

Deep-sea-derived butyrolactone I (BTL-I), which was identified as a type of butanolide, was isolated from Aspergillus sp. Ovalbumin (OVA)-induced BALB/c anaphylaxis was established to explore the antifood allergic activity of BTL-I. As a result, BTL-I was able to alleviate OVA-induced allergy symptoms, reduce the levels of histamine and mouse mast cell proteinases, inhibit OVA-specific IgE, and decrease the population of mast cells in the spleen and mesenteric lymph nodes. BTL-I also significantly suppressed mast-dependent passive cutaneous anaphylaxis. Additionally, the maturation of bone marrow-derived mast cells (BMMCs) declined as BTL-I caused down-regulation of c-KIT receptors. Furthermore, molecular docking analyses revealed that BTL-I interacted with the inhibitory receptor, FcγRIIB. In conclusion, the reduction of mast cell function by deep-sea-derived BTL-I as well as its interactions with the inhibitory receptor, FcγRIIB, may contribute to BTL-I-related protection against food anaphylaxis.

Topics: 4-Butyrolactone; Anaphylaxis; Animals; Aspergillus; Cells, Cultured; Disease Models, Animal; Female; Food Hypersensitivity; Histamine; Humans; Immunoglobulin E; Mast Cells; Mice; Mice, Inbred BALB C; Ovalbumin; Proto-Oncogene Proteins c-kit; Seawater

Cry1Ac toxin, from Bacillus thuringiensis, is widely used as a biopesticide and expressed in genetically modified (GM) plants used for human and animal consumption. Since Cry1Ac is also immunogenic and able to activate macrophages, it is crucial to thoroughly evaluate the immunological effects elicited after intra-gastric administration. The allergenic potential of purified Cry1Ac was assessed and compared with that induced in a murine model of food-allergy to ovalbumin (OVA), in which animals are sensitized with the adjuvant Cholera toxin (CT). Mice were weekly intragastrically administered with: i) vehicle phosphate-buffered saline (PBS), ii) OVA, iii) OVA plus CT iv) Cry1Ac or v) OVA plus Cry1Ac. Seven weeks after, mice were intragastrically challenged and allergic reactions along with diverse allergy related immunological parameters were evaluated at systemic and intestinal level. The groups immunized with, Cry1Ac, OVA/Cry1Ac or OVA/CT developed moderate allergic reactions, induced significant IgE response and increased frequencies of intestinal granulocytes, IgE+ eosinophils and IgE+ lymphocytes. These same groups also showed colonic lymphoid hyperplasia, notably in humans, this has been associated with food allergy and intestinal inflammation. Although the adjuvant and allergenic potential of CT were higher than the effects of Cry1Ac, the results show that applied intra-gastrically at 50 μg doses, Cry1Ac is immunogenic, moderately allergenic and able to provoke intestinal lymphoid hyperplasia. Moreover, Cry1Ac is also able to induce anaphylaxis, since when mice were intragastrically sensitized with increasing doses of Cry1Ac, with every dose tested, a significant drop in rectal temperature was recorded after intravenous challenge.

Topics: Allergens; Anaphylaxis; Animals; Bacillus thuringiensis; Bacillus thuringiensis Toxins; Bacterial Proteins; Disease Models, Animal; Endotoxins; Female; Food Hypersensitivity; Hemolysin Proteins; Humans; Immunization; Immunoglobulin E; Inflammation; Intestines; Mice; Mice, Inbred BALB C; Ovalbumin; Pest Control, Biological; Plants, Genetically Modified

Topics: Acid Phosphatase; Animals; Enzyme-Linked Immunosorbent Assay; Female; Food Hypersensitivity; Immunoglobulin E; Immunoglobulin G; Injections, Intraperitoneal; Mice, Inbred BALB C; Milk Hypersensitivity; Milk Proteins; Ovalbumin; Plant Roots; Solanum tuberosum; Time Factors

Our diet is known to substantially influence the immune response not only by support of mucosal barriers but also via direct impact on immune cells. Thus, it was of great interest to compare the immunological effect of two mouse chows with substantial differences regarding micro-, macronutrient, lipid and vitamin content on the food allergic response in our previously established mouse model. As the two mouse chows of interest, we used a soy containing feed with lower fatty acid (FA) amount (soy-containing feed) and compared it to a soy free mouse chow (soy-free feed) in an established protocol of oral immunizations with Ovalbumin (OVA) under gastric acid suppression. In the animals receiving soy-containing feed, OVA-specific IgE, IgG1, IgG2a antibody levels were significantly elevated and food allergy was evidenced by a drop of body temperature after oral immunizations. In contrast, mice on soy-free diet had significantly higher levels of IL-10 and were protected from food allergy development. In conclusion, soy-containing feed was auxiliary during sensitizations, while soy-free feed supported oral tolerance development and food allergy prevention.

Topics: Animal Feed; Animals; Body Temperature; Disease Models, Animal; Fatty Acids; Female; Food Hypersensitivity; Immune Tolerance; Immunization; Immunoglobulin A; Immunoglobulin E; Immunoglobulin G; Interleukin-10; Mice; Mice, Inbred BALB C; Ovalbumin; Soy Foods; Soybean Proteins

Processed foods are part of daily life. Almost all processed foods contain food additives such as sweeteners, preservatives and colourants. From childhood, it is difficult to avoid consuming food additives. It is thought that oral tolerance for food antigens is acquired during early life. If tolerance fails, adverse immune responses to food proteins may occur.. We hypothesized that food additives prevent acquisition of oral tolerance and aimed to verify the safety of food additives.. We induced experimental oral tolerance in mice for ovalbumin (OVA), a food antigen, by previous oral treatment with OVA before sensitization with OVA injections. Food additives were administered at the induction of oral tolerance, and food allergy was induced by repeated administration of OVA. Symptoms of food allergy were defined as a change in body temperature and allergic diarrhoea.. Saccharin sodium and a mixture of food additives inhibited acquisition of oral tolerance. Hypothermia and allergic diarrhoea with elevation of OVA-specific IgE were induced in the murine model of oral tolerance. Analyses of antigen-presenting cells in mesenteric lymph nodes showed that food additives affected their manner of migration. Additionally, food additives decreased the proportion of CD25. A large amount of food additives may prevent acquisition of oral tolerance. Intake of food additives in early life may increase the risk of food allergies.

Topics: Administration, Oral; Allergens; Animals; Antigen-Presenting Cells; Biomarkers; Chemotaxis; Disease Models, Animal; Enzyme-Linked Immunosorbent Assay; Female; Food Additives; Food Hypersensitivity; Immune Tolerance; Immunoglobulin E; Immunoglobulin G; Mice; Ovalbumin; Saccharin; Sweetening Agents

To determine whether oral administration of. Ovalbumin (OVA)-induced allergic asthma and β-lactoglobulin-induced food allergy mouse models were used in this study. Following oral administration of. Oral administration of

Topics: Allergens; Animals; Asthma; Bifidobacterium longum subspecies infantis; Bronchoalveolar Lavage Fluid; Cytokines; Disease Models, Animal; Food Hypersensitivity; Immunoglobulin E; Immunoglobulin G; Inflammation; Interleukin-13; Interleukin-4; Intestines; Lactoglobulins; Male; Mice; Mice, Inbred BALB C; Ovalbumin; Probiotics

Calprotectin, also known as S100A8/A9, has been linked to gut inflammation caused by IgE-mediated food hypersensitivities, but the pathophysiologic abnormalities it causes remain to be determined. We created a mild food hypersensitivity model through oral gavage of ovalbumin in Norway brown rats without using immune adjuvant. Changes in the levels of calprotectin and inflammation-associated cytokines were then observed over time. We found that fecal calprotectin as well as jejunal and liver TLR4, TNF-α, NF-κB, IL-1β, and IL-6 were upregulated in hypersensitive rats. Additionally, the influence of calprotectin on CD4+ T and dendritic cells was observed by co-culturing CD4+ T cells with dendritic cells, which revealed a shift toward increased Th2 T cells in calprotectin-treated cultures. These results suggest that calprotectin, along with other inflammatory factors, promotes the inflammation seen in mild food allergy.

Topics: Allergens; Animals; CD4-Positive T-Lymphocytes; Cells, Cultured; Cytokines; Dendritic Cells; Disease Models, Animal; Food Hypersensitivity; Humans; Immunoglobulin E; Inflammation; Inflammation Mediators; Leukocyte L1 Antigen Complex; Lymphocyte Activation; Male; Ovalbumin; Rats; Rats, Inbred BN; Th1-Th2 Balance; Up-Regulation

To investigate the effects of adipose-derived stem cells (ADSC) and non-methylated CpG-oligodeoxynucleotides (CpG-ODN) on the expression of peripheral blood CD4. A total of 40 female BALB/c mice were randomly divided into control group, allergic group, ADSC treatment group, and CpG-ODN treatment group, with 10 mice in each group. A mouse model of food allergy was established by intraperitoneal injection and intragastric administration of ovalbumin (OVA) for sensitization and challenge. The mice in the control group were treated with normal saline at the same dose; the mice in the ADSC treatment group were given intraperitoneal injection of ADSC (1×10. The allergic group had significantly higher allergic symptom scores and serum level of OVA-IgE than the control group (P<0.05). There were no significant differences in the allergic symptom score and the serum level of OVA-IgE between the ADSC treatment group and the CpG-ODN treatment group (P>0.05), but these two groups had significantly lower allergic symptom scores and serum level of OVA-IgE than the allergic group and significantly higher allergic symptom scores and serum level of OVA-IgE than the control group (P<0.01). The allergic group had a significantly lower percentage of peripheral blood CD4. ADSC and non-methylated CpG-ODN have a certain effect in the treatment of food allergy and can increase the percentage of peripheral blood CD4

Topics: Adipose Tissue; Adjuvants, Immunologic; Animals; Female; Food Hypersensitivity; Immunoglobulin E; Mice; Mice, Inbred BALB C; Oligodeoxyribonucleotides; Ovalbumin; Stem Cell Transplantation; T-Lymphocytes, Regulatory

Currently, there are no efficient medications available for the prevention and treatment of food allergy (FA). Herbal medicines, including traditional Japanese Kampo medicines (TJKMs), are promising therapeutic drugs.. We screened 18 TJKMs for treatment of FA symptoms in a mouse FA model induced by ovalbumin (OVA). BALB/c mice were sensitized intraperitoneally by an OVA/aluminum hydroxide gel mixture followed by 4 booster doses of oral OVA and FA symptom induction by 50 mg of OVA. TJKMs were orally administered for 28 days from the day of sensitization to the day before FA symptom induction. Evaluated FA symptoms included a decrease in body temperature and allergic diarrhea. Allergic sensitization was determined by plasma OVA-specific IgE levels. Cytokine mRNA levels in mesenteric lymph nodes, plasma mouse mast cell protease-1, and the number of mast cells in the small and large intestines were analyzed. Additionally, the therapeutic effect of the TJKM eppikajutsuto (EJT) on mast cell degranulation was determined in active anaphylaxis and passive cutaneous anaphylaxis models.. EJT effectively prevented FA symptoms. Although OVA-specific IgE levels and the intestinal mast cell numbers were not different between the EJT-treated and untreated FA mice, plasma mMcpt1 and IL-4 levels were lower in EJT-treated FA mice than untreated FA mice. EJT could alleviate symptoms in both active and passive anaphylaxis models.. EJT prevented OVA-induced FA symptoms in a mouse model, suggesting that EJT might exert its therapeutic activity via IL-4 suppression and the inhibition of mucosal mast cell degranulation.

Topics: Allergens; Anaphylaxis; Animals; Anti-Allergic Agents; Cell Degranulation; Cytokines; Disease Models, Animal; Female; Food Hypersensitivity; Immunoglobulin E; Intestinal Mucosa; Male; Mast Cells; Medicine, Kampo; Mice, Inbred BALB C; Mice, Inbred ICR; Ovalbumin; Peptide Hydrolases; Pharmaceutical Preparations; Plant Extracts; RNA, Messenger

Atopic dermatitis (AD) is a common inflammatory skin disease with dysfunction of the skin barrier, an abnormal immune response and frequent allergies to environmental antigens like food antigens. Clinical observations suggest that certain diets can influence the course of AD.. Here we compared the phenotype of food allergen-specific T cells activated through skin or gut allergen exposure to transfer skin inflammation into naïve recipients upon epicutaenous allergen challenge.. TCR-transgenic T cells activated through epicutaneous or oral OVA exposure both migrate to skin lymph nodes after adoptive transfer and epicutaenous OVA exposure. AD-like skin inflammation could only be induced by the transfer of epicutaneously primed OVA T cells. Analysis of the immune phenotype demonstrated an IL-22/IL-17A-dominated immune phenotype of skin-pathogenic T cells.. IL-22 seems to be the critical cytokine for the development of AD and is induced in this model by epicutaneous sensitization with OVA.

Topics: Allergens; Animals; CD4-Positive T-Lymphocytes; Dermatitis, Atopic; Disease Models, Animal; Female; Food Hypersensitivity; Humans; Interleukin-17; Interleukin-22; Interleukins; Intestines; Mice; Mice, Inbred BALB C; Mice, Transgenic; Ovalbumin; Skin; Vaccination

Oral immunotherapy (OIT) is a promising treatment of food allergy. To administer an appropriate oral dose of an allergenic component as OIT to individuals sensitized with a food allergen may prevent inducing food allergic inflammation in them. So we attempted to establish a mouse model to evaluate efficacy for oral administration of food allergen after sensitization. In BALB/c mice sensitized by injecting ovalbumin (OVA) with alum twice, OVA was administered before inducing inflammation by feeding the mice with egg white (EW) diet. Severe inflammatory responses, such as enteropathy, weight loss, IL-4 production, and increase of IgE antibody levels, were suppressed by administration with 4 mg of OVA 7 times before feeding EW diet. OVA administration alone induced a slight Th2 response, but no symptoms. The current study demonstrated that severe food allergic enteropathy could be prevented by pre-administration with appropriate dose of OVA to sensitized mice.

Topics: Administration, Oral; Animals; Disease Models, Animal; Dose-Response Relationship, Drug; Female; Food Hypersensitivity; Immunization; Immunoglobulin E; Interleukin-4; Intestinal Diseases; Mice; Mice, Inbred BALB C; Ovalbumin

The therapeutic effect of baicalin and its mechanism were explored.. A total of 30 Sprague Dawley (SD) rats were randomly divided into 3 groups of 10: ovalbumin group (OVA group), baicalin intervention group (HQ group), and saline-group (NC group). Serum OVA-IgE antibody levels were detected by enzyme-linked immunosorbent assay (ELISA); and diarrhea in rats was observed. Animals were sacrificed at week seven. Then, a 5-cm long duodenum beneath the Treitz ligament was collected from each rat, and was fixed, embedded, sliced and stained with toluidine blue to evaluate the integrity of mast cells. Next, pathological changes of the intestine were observed by hematoxylin and eosin (H&E) staining, and the ultrastructure of the intestinal mucosa was observed under a transmission electron microscope.. Serum OVA-sIgE level were significantly lower (at sixth week, OVA group: 12.86±1.35, HQ group: 3.47±0.51, F=117.05, P<0.01), the number of eosinophils significantly decreased (HQ group: 2.73±1.02, OVA group: 16.48±2.32, P<0.01), mast cell integrated rate was significantly increased (HQ group: 89.90±4.43, OVA group: 35.30±9.78, P<0.01) uniform small intestinal villi were observed, the organelles were basically normal, and lesions were significantly fewer in the HQ group, compared with the OVA group.. Baicalin can effectively reduce serum OVA-sIgE in rats with food allergy, increase mast cell integrated rate and alleviate intestinal pathological changes. Hence, baicalin has a good therapeutic effect on food allergy.

Topics: Animals; Anti-Inflammatory Agents, Non-Steroidal; Disease Models, Animal; Female; Flavonoids; Food Hypersensitivity; Immunoglobulin E; Intestinal Mucosa; Intestine, Small; Male; Mast Cells; Ovalbumin; Protective Agents; Rats, Sprague-Dawley

The purpose of this study was to screen phytochemicals capable of inducing immune tolerance via enhanced transforming growth factor-β1 (TGF-β1) secretion and investigate their effects in a mouse model of food allergy and colitis. In a screening test using THP-1-derived dendritic cells, a significant increase in TGF-β1 levels was observed upon treatment with ferulic acid and its glycosides, among which ferulic acid rutinoside (FAR) induced the highest level of TGF-β1 secretion. Oral administration of FAR suppressed serum levels of immunoglobulin E and histamine in ovalbumin-sensitized mice and triggered the differentiation of regulatory T (Treg) cells. In comparison to the control, FAR treatment also induced stronger TGF-β1 secretion from splenic dendritic cells. FAR treatment attenuated dextran-sulfate-sodium-induced colitis in the model mice and induced Treg differentiation. These results suggest that FAR exerts potent immunomodulatory effects against allergic and intestinal inflammatory responses by inducing Treg differentiation.

Topics: Animals; Chromatography, High Pressure Liquid; Colitis; Coumaric Acids; Dendritic Cells; Dextran Sulfate; Disease Models, Animal; Female; Food Hypersensitivity; Glycosides; Histamine; Humans; Immune Tolerance; Immunoglobulin E; Lymphocyte Activation; Mice; Mice, Inbred BALB C; Ovalbumin; T-Lymphocytes, Regulatory; Transforming Growth Factor beta1; Up-Regulation

Fructo-oligosaccharides (FOS) are prebiotic agents with immunomodulatory effects involving improvement of the intestinal microbiota and metabolome. In this study, we investigated the cellular mechanisms through which FOS modulate intestinal antigen-specific CD4+ T cell responses in food allergy, using OVA23-3 mice.. OVA23-3 mice were fed an experimental diet containing either ovalbumin (OVA) or OVA and FOS for 1 week. Body weight and mucosal mast cell protease 1 in the serum were measured as the indicator of intestinal inflammation. Single-cell suspensions were prepared from intestinal and systemic lymphoid tissues for cellular analysis. Cytokine production was measured by ELISA. Activation markers and intracellular cytokines in CD4+ T cells were analyzed by flow cytometry. Activated CD4+ T cells were purified to examine cytokine production.. Dietary intake of FOS provided moderate protection from the intestinal inflammation induced by the OVA-containing diet. FOS significantly reduced food allergy-induced Th2 cytokine responses in intestinal tissues but not in systemic tissues. FOS decreased OVA diet-induced IFN-γ+IL-4+ double-positive CD4+ T cells and early-activated CD45RBhighCD69+CD4+ T cells in the mesenteric lymph nodes. Furthermore, we confirmed that these CD45RBhighCD69+CD4+ T cells are able to produce high levels of IFN-γ and moderate level of IL-4, IL-10, and IL-13.. Dietary intake of FOS during the development of food allergy attenuates the induction of intestinal Th2 cytokine responses by regulating early activation of naïve CD4+ T cells, which produce both Th1 and Th2 cytokines. Our results suggest FOS might be a potential food agent for the prevention of food allergy by modulating oral sensitization to food antigens.

Topics: Animals; Cells, Cultured; Cytokines; Diet; Disease Models, Animal; Female; Food Hypersensitivity; Fructose; Intestines; Lymphocyte Activation; Lymphoid Tissue; Mice; Mice, Inbred BALB C; Mice, Transgenic; Oligosaccharides; Ovalbumin; Th1 Cells; Th2 Cells

Healthy foods like polyphenol-rich berries and high quality edible proteins are in demand in today's functional food marketplace, but it can be difficult to formulate convenient food products with physiologically-relevant amounts of these ingredients and still maintain product quality. In part, this is because proteins can interact with other food ingredients and precipitate destabilizing events, which can disrupt food structure and diminish shelf life. Proteins in foods can also interact with human receptors to provoke adverse consequences such as allergies. When proteins and polyphenols were pre-aggregated into stable colloidal particles prior to use as ingredients, highly palatable food formulations (with reduced astringency of polyphenols) could be prepared, and the overall structural properties of food formulations were significantly improved. All of the nutritive and phytoactive benefits of the proteins and concentrated polyphenols remained highly bioavailable, but the protein molecules in the particle matrix did not self-aggregate into networks or react with other food ingredients. Both the drainage half-life (a marker of structural stability) and the yield stress (resistance to flow) of model foams made with the protein-polyphenol particles were increased in a dose-dependent manner. Of high significance in this complexation process, the reactive allergenic epitopes of certain proteins were effectively blunted by binding with polyphenols, attenuating the allergenicity of the food proteins. Porcine macrophages produced TNF-α proinflammatory cytokine when provoked with whey protein, but, this response was blocked completely when the cells were stimulated with particles that complexed whey protein with cinnamon-derived polyphenols. Cytokine and chemokine production characteristic of allergic reactions were blocked by the polyphenols, allowing for the potential creation of hypoallergenic protein-berry polyphenol enriched foods.

Topics: Animals; Dendritic Cells; Food Hypersensitivity; Fruit; Fruit and Vegetable Juices; Functional Food; Humans; Macrophages; Ovalbumin; Plant Extracts; Polyphenols; Swine; Tumor Necrosis Factor-alpha; Whey Proteins

Food allergy is immediate hypersensitive reactions to ingested foods. Since early diagnosis is effective for disease control, development of an objective diagnostic index is required. Using mediator-lipidomics, we found that levels of the urinary prostaglandin D

Topics: Animals; Asthma; Dermatitis, Atopic; Food Hypersensitivity; Humans; Hyperplasia; Intestines; Intramolecular Oxidoreductases; Lipocalins; Mast Cells; Mice; Mice, Inbred BALB C; Ovalbumin; Prostaglandin D2; Rhinitis, Allergic

The attempt to induce oral tolerance as a treatment for food allergy has been hampered by a lack of sustained clinical protection. Immunotherapy by nonoral routes, such as the skin, may be more effective for the development of maintained tolerance to food allergens.. We sought to determine the efficacy and mechanism of tolerance induced by epicutaneous immunotherapy (EPIT) in a model of food-induced anaphylaxis.. C3H/HeJ mice were sensitized to ovalbumin (OVA) orally or through the skin and treated with EPIT using OVA-Viaskin patches or oral immunotherapy using OVA. Mice were orally challenged with OVA to induce anaphylaxis. Antigen-specific regulatory T (Treg)-cell induction was assessed by flow cytometry using a transgenic T-cell transfer model.. By using an adjuvant-free model of food allergy generated by epicutaneous sensitization and reactions triggered by oral allergen challenge, we found that EPIT induced sustained protection against anaphylaxis. We show that the gastrointestinal tract is deficient in de novo generation of Treg cells in allergic mice. This defect was tissue-specific, and epicutaneous application of antigen generated a population of gastrointestinal-homing LAP. Our data highlight the immune communication between skin and gastrointestinal tract, and identifies novel mechanisms by which epicutaneous tolerance can suppress food-induced anaphylaxis.

Topics: Administration, Cutaneous; Allergens; Anaphylaxis; Animals; Arachis; Cholera Toxin; Desensitization, Immunologic; Food Hypersensitivity; Forkhead Transcription Factors; Immunoglobulin G; Mice, Inbred BALB C; Mice, Inbred C3H; Ovalbumin; Peptides; T-Lymphocytes, Regulatory; Transforming Growth Factor beta

Mucosal mast cells (MMCs) play a central role in the development of symptoms associated with IgE-mediated food allergy. Recently, Notch2-mediated signaling was shown to be involved in proper MMC distribution in the intestinal tract.. This study aimed to clarify the mechanism by which Notch signaling regulates MMC distribution in the intestinal mucosa. Furthermore, pharmacologic inhibition of Notch signaling was evaluated as a treatment for symptoms associated with experimental food allergy.. Bone marrow-derived mast cells generated from mice were cultured with Notch ligands, and then expression of genes associated with MMCs was measured in the cells. In addition, the effect of an inhibitor of Notch signaling on food antigen-induced allergic reactions was examined in a mouse model of food allergy.. Notch signaling induced MMC differentiation through upregulation of expression of genes characteristic of MMCs in the presence of IL-3. Some lamina propria cells isolated from the mouse small intestine expressed Notch ligands and were able to upregulate MMC markers in bone marrow-derived mast cells through Notch signaling. In a mouse model of food allergy, administration of a Notch signaling inhibitor led to suppression of food antigen-induced hyperplasia of intestinal MMCs, resulting in alleviation of allergic diarrhea and systemic anaphylaxis.. Notch signaling contributes to differentiation and accumulation of MMCs in the intestinal mucosa. Thus inhibition of Notch signaling alleviates symptoms associated with experimental food allergy. These results raise the possibility that Notch signaling in mast cells is a novel target for therapy in patients with food allergy.

Topics: Allergens; Animals; Cytokines; Dipeptides; Female; Food Hypersensitivity; Hyperplasia; Intestinal Mucosa; Intestine, Small; Mast Cells; Mice, Inbred BALB C; Ovalbumin; Receptors, Notch; Signal Transduction

Oral tolerance induction in early life is a promising approach for food allergy prevention. Its success requires the identification of factors necessary for its persistence.. We aimed to assess in mice duration of allergy prevention by breastfeeding-induced oral tolerance and whether oral TGF-β supplementation after weaning would prolong it.. We quantified ovalbumin (OVA) and OVA-specific immunoglobulin levels by ELISA in milk from the EDEN birth cohort. As OVA-specific Ig was found in all samples, we assessed whether OVA-immunized mice exposed to OVA during lactation could prevent allergic diarrhoea in their 6- and 13-week-old progeny. In some experiments, a TGF-β-enriched formula was given after weaning.. At 6 weeks, only 13% and 34% of mice breastfed by OVA-exposed mothers exhibited diarrhoea after six and seven OVA challenges vs. 44% and 72% in mice breastfed by naïve mothers (P = 0.02 and 0.01). Protection was associated with decreased levels of MMCP1 and OVA-specific IgE (P < 0.0001). At 13 weeks, although OVA-specific IgE remained low (P = 0.001), diarrhoea occurrence increased to 32% and 46% after six and seven OVA challenges in mice breastfed by OVA-exposed mothers. MMCP1 levels were not significantly inhibited. Supplementation with TGF-β after weaning induced a strong protection in 13-week-old mice breastfed by OVA-exposed mothers compared with mice breastfed by naive mothers (0%, 13% and 32% of diarrhoea at the fifth, sixth and seventh challenges vs. 17, 42 and 78%; P = 0.05, 0.0043 and 0.0017). MMCP1 levels decreased by half compared with control mice (P = 0.02). Prolonged protection was only observed in mice rendered tolerant by breastfeeding and was associated with an improved gut barrier.. In mice, prevention of food allergy by breastfeeding-induced tolerance is of limited duration. Nutritional intervention by TGF-β supplementation after weaning could prolong beneficial effects of breast milk on food allergy prevention.

Topics: Animal Feed; Animals; Antibody Specificity; Breast Feeding; Diarrhea; Food Hypersensitivity; Humans; Immune Tolerance; Immunoglobulin E; Immunoglobulin G; Intestinal Mucosa; Mice; Milk, Human; Ovalbumin; T-Lymphocyte Subsets; Transforming Growth Factor beta; Weaning

The prevalence of food allergies worldwide has increased recently. Epicutaneous sensitization to antigen could be a method to study food allergy. To clarify the mechanisms of food allergy, we established a mouse model of epicutaneous sensitization using ovalbumin (OVA). BALB/c mice were sensitized by three-time application of OVA to tape-stripped skin (1-week sensitization at 2-week intervals) and oral challenge of OVA undertaken. Rectal temperature was monitored. Blood and tissue (skin and jejunum) of challenged mice were taken. Numbers of mast cells (MCs) and basophils were counted. Serum and/or tissue levels of OVA -specific IgE and IgG antibodies and several cytokines were measured using enzyme-linked immunoassay kits. MC and basophil depletion experiments were undertaken. In OVA/epicutaneous-sensitized and orally challenged mice, systemic anaphylaxis (as evidenced by reduced rectal temperature) was observed. Levels of OVA-specific IgE and IgG antibodies were increased in these mice, as were increased number of MCs and basophils. Serum levels of MC protease 1 were increased significantly. Basophil and MC depletion experiments revealed that they both participate in reactions. Increased production of thymic stromal lymphopoietin (TSLP) at skin sites of OVA sensitization was noted. We speculate that TSLP produced from epidermal cells during antigen sensitization can enable basophils to promote a T helper (Th)2 immune reaction, leading to and systemic anaphylaxis by antigen-specific IgE-bearing MCs. This TSLP-basophils-MC axis could be a novel therapeutic target against food allergy.

Topics: Anaphylaxis; Animals; Basophils; Cytokines; Food Hypersensitivity; Jejunum; Mast Cells; Mice, Inbred BALB C; Ovalbumin; Skin; Thymic Stromal Lymphopoietin

Previous studies have attributed to the cocoa powder the capacity to attenuate the immune response in a rat oral sensitization model. To gain a better understanding of cocoa-induced mechanisms at small intestinal level, 3-week-old female Lewis rats were fed either a standard diet or a diet containing 10% cocoa for 4 weeks with or without concomitant oral sensitization with ovalbumin (OVA). Thereafter, we evaluated the lymphocyte composition of the Peyer's patches (PPL), small intestine epithelium (IEL) and lamina propria (LPL). Likewise, gene expression of several immune molecules was quantified in the small intestine. Moreover, histological samples were used to evaluate the proportion of goblet cells, IgA+ cells and granzyme+cells as well. In cocoa-fed animals, we identified a five-time reduction in the percentage of IgA+ cells in intestinal tissue together with a decreased proportion of TLR4+ IEL. Analyzing the lymphocyte composition, almost a double proportion of TCRγδ+cells and an increase of NK cell percentage in PPL and IEL were found. In addition, a rise in CD25+, CD103+ and CD62L- cell proportions was observed in CD4+ PPL from cocoa-fed animals, along with a decrease in gene expression of CD11b, CD11c and IL-10. These results suggest that changes in PPL and IEL composition and in the gene expression induced by the cocoa diet could be involved, among other mechanisms, on its tolerogenic effect.

Topics: Administration, Oral; Animals; Cacao; Disease Models, Animal; Epithelium; Female; Food Hypersensitivity; Immunoglobulin A; Intestine, Small; Lymphocytes; Lymphoid Tissue; Ovalbumin; Peyer's Patches; Rats, Inbred Lew; Receptors, Antigen, T-Cell, gamma-delta; Toll-Like Receptor 4

The mechanism inducing either inflammation or tolerance to orally administered food allergens remains unclear. To investigate this we analyzed mouse models of food allergy (OVA23-3) and tolerance (DO11.10 [D10]), both of which express ovalbumin (OVA)-specific T-cell receptors.. OVA23-3, recombination activating gene (RAG)-2-deficient OVA23-3 (R23-3), D10, and RAG-2-deficient D10 (RD10) mice consumed a diet containing egg white (EW diet) for 2-28 days. Interleukin (IL)-4 production by CD4+ T cells was measured as a causative factor of enteropathy, and anti-IL-4 antibody was used to reveal the role of Foxp3+ OVA-specific Tregs (aiTreg) in this process.. Unlike OVA23-3 and R23-3 mice, D10 and RD10 mice did not develop enteropathy and weight loss on the EW diet. On days 7-10, in EW-fed D10 and RD10 mice, splenic CD4+ T cells produced significantly more IL-4 than did those in the mesenteric lymph nodes (MLNs); this is in contrast to the excessive IL-4 response in the MLNs of EW-fed OVA23-3 and R23-3 mice. EW-fed R23-3 mice had few aiTregs, whereas EW-fed RD10 mice had them in both tissues. Intravenous injections of anti-IL-4 antibody recovered the percentage of aiTregs in the MLNs of R23-3 mice. On day 28, in EW-fed OVA23-3 and R23-3 mice, expression of Foxp3 on CD4+ T cells corresponded with recovery from inflammation, but recurrence of weight loss was observed on restarting the EW diet after receiving the control-diet for 1 month. No recurrence developed in D10 mice.. Excessive IL-4 levels in the MLNs directly inhibited the induction of aiTregs and caused enteropathy. The aiTregs generated in the attenuation of T cell-dependent food allergic enteropathy may function differently than aiTregs induced in a tolerance model. Comparing the two models enables to investigate their aiTreg functions and to clarify differences between inflammation with subsequent desensitization versus tolerance.

Topics: Allergens; Animals; Desensitization, Immunologic; Female; Food Hypersensitivity; Immune Tolerance; Interleukin-4; Intestinal Mucosa; Lymph Nodes; Mice; Ovalbumin; Receptors, Antigen, T-Cell; T-Lymphocytes, Regulatory

Immunoglobulin E (IgE)-mediated food allergy, such as egg white allergy, is common in young children (<3 years old), but not all young children sensitive to egg white present with allergic symptoms. This study investigated the relationship between sensitization to egg white component allergens and clinical manifestations of allergic diseases in young children.. From March to December 2010, 2256 children with physician-diagnosed allergic diseases were tested for serum levels of egg white, ovalbumin, and ovomucoid-specific IgE in the Pediatric Allergy and Asthma Center of Chang Gung Memorial Hospital. Serum was analyzed for specific IgE antibodies to egg white, ovalbumin, and ovomucoid by ImmunoCAP (Phadia, Uppsala, Sweden). Allergen-specific IgE levels ≥0.35 kUA/L were defined as positive.. There was a significantly higher sensitization rate to egg white and its components in children aged 2-4 years old. The sensitization rate to egg white, ovalbumin, and ovomucoid in this age group was 53.5%, 48.3%, and 37.2%, respectively, and the trend of the sensitization decreased with age (p < 0.001). After adjusting for age, sensitization to egg white and ovalbumin was associated with children with dermatitis [egg white: odds ratio (OR) = 1.28, 95% confidence intervals (CI) = 1.03-1.58, p < 0.05; ovalbumin: OR = 1.30, 95% CI = 1.04-1.62, p < 0.05]. Children with ovomucoid sensitization had no statistically significant risk among different groups in the current study.. Children aged 2-4 years old have higher sensitivity to egg white, ovalbumin, and ovomucoid. Children with egg white and ovalbumin sensitization have a higher risk for atopic dermatitis, and ovalbumin has a more important contribution. Furthermore, we suggested that in children with atopic dermatitis, if they are aged 2-4 years old and are having egg white and ovalbumin sensitization, avoiding eating raw or slightly heated eggs might have a beneficial effect.

Topics: Adolescent; Age Factors; Allergens; Child; Child, Preschool; Egg White; Female; Food Hypersensitivity; Humans; Immunoassay; Immunoglobulin E; Infant; Male; Ovalbumin; Sweden

Allergic mice show a reduction in body weight and adiposity with a higher inflammatory response in the adipose tissue similar to obese fat tissue. This study aimed to evaluate whether the low-grade inflammatory milieu of mice with diet-induced mild obesity interferes with the allergic response induced by ovalbumin (OVA).. BALB/c mice were divided into four groups: 1) non-allergic (OVA-) mice fed chow diet, 2) allergic (OVA+) mice fed chow diet, 3) OVA- mice fed high-refined carbohydrate-containing (HC) diet, and 4) OVA+ mice fed HC diet. After 5 wk, allergic groups were sensitized with OVA and received a booster 14 d later. All groups received an oral OVA challenge 7 d after the booster.. Allergic groups showed increased serum levels of total IgE, anti-OVA IgE, and IgG1; a high disease activity index score; aversion to OVA; and increased intestinal eosinophil infiltration. Non-allergic mild-obese mice also showed aversion to OVA and an increased number of eosinophils in the proximal jejunum. After the allergic challenge, OVA+ mice fed chow diet showed weight loss and lower adiposity in several adipose tissue depots. OVA+ mice fed HC diet showed a loss of fat mass only in the mesenteric adipose tissue. Furthermore, increased levels of TNF, IL-6, and IL-10 were observed in this tissue.. Our data show that mild-obese allergic mice do not present severe pathologic features of food allergy similar to those exhibited by lean allergic mice. Mild obesity promoted by HC diet ingestion causes important intestinal disorders that appear to modulate the inflammatory response during the antigen challenge.

Topics: Adipose Tissue; Adiposity; Animals; Body Weight; Diet; Dietary Carbohydrates; Food Hypersensitivity; Glucose Tolerance Test; Immunoglobulin E; Immunoglobulin G; Inflammation; Insulin Resistance; Interleukin-10; Interleukin-6; Intestinal Mucosa; Intestines; Leukocyte Count; Male; Mice; Mice, Inbred BALB C; Obesity; Ovalbumin

The role of maternal allergen exposure in the allergenicity of the offspring remains controversial. Some studies have shown that maternal exposure is a risk factor for allergy in the offspring, whereas other studies have shown that maternal exposure induces immune tolerance and protects offspring from allergy disease. Therefore, we utilized maternal rat allergen exposure model to evaluate the offspring immune reactions to ovalbumin protein and to determine whether the Brown Norway (BN) rat model is a suitable animal model for studying the allergenicity of food proteins. For three generations, rats received an allergens or non-allergens by gavage during the pregnancy and lactation periods. After weaning, the offspring rats were used for oral sensitization experiment. In the sensitization experiment, the control rat, which had maternal exposure to phosphate-buffered saline (PBS), exhibited full response of IgG to oral exposure to OVA. The IgG level was significantly lower in F1 rats that were sensitized by maternal exposure to ovalbumin(OVA). Moreover, the lowest IgG level was found for the F3b sensitized by maternal rats exposed to OVA allergen for three continuous generations. Compared with maternal OVA exposure prior to postnatal sensitization, the sensitization via maternal PBS led to a higher serum level of OVA-specific IgG. However, the OVA-specific IgG levels for the two generations of maternal PBS exposure prior to postnatal sensitization was not higher than that for the one generation of maternal rats exposed to PBS prior to postnatal sensitization. Our studies demonstrate that maternal OVA exposure during the pregnancy and lactation can affect the results of oral sensitization studies using ovalbumin protein. BN rats must be bred in non-allergen conditions for at least one generation to avoid problems in rat models for studying the allergenicity of food proteins.

Topics: Administration, Oral; Allergens; Animals; Antibodies; Disease Models, Animal; Female; Food Hypersensitivity; Immunization; Immunoglobulin E; Immunoglobulin G; Maternal Exposure; Ovalbumin; Pregnancy; Prenatal Exposure Delayed Effects; Rats; Rats, Inbred BN

TIM4 (T cell immunoglobulin mucin domain molecule-4) plays a critical role in the initiation of skewed T helper (Th) 2 polarization. The factors regulating TIM4 expression are unclear. This study tests a hypothesis that p300 and STAT6 (signal transducer and activator transcription-6) regulates TIM4 expression in dendritic cells (DC). In this study, a food allergy mouse model was developed with ovalbumin (a specific antigen) and cholera toxin (CT; an adjuvant). The chromatin immunoprecipitation assay was performed to evaluate the chromatin changes at TIM4 and STAT6 promoters. The TIM4 expression was evaluated by real time RT-PCR and Western blotting. The results showed that high levels of p300 and TIM4 were detected in the intestinal DCs of mice with intestinal allergy. p300 is involved in the CT-induced TIM4 expression in DCs. p300 interacts with the chromatin at the TIM4 promoter locus in DCs isolated from allergic mice. CT increases p300 expression to regulate STAT6 levels in DCs. STAT6 mediates the CT-induced TIM4 expression in DCs. In conclusion, p300 and STAT6 mediate the microbial product CT-induced TIM4 expression in DCs.

Topics: Animals; Cell Polarity; Cholera Toxin; Dendritic Cells; Disease Models, Animal; Food Hypersensitivity; Gene Expression Regulation; Humans; Intestinal Mucosa; Intestines; Membrane Proteins; Mice; Ovalbumin; p300-CBP Transcription Factors; Promoter Regions, Genetic; STAT6 Transcription Factor; Th2 Cells

The objective of this study was to determine the effect of feeding a maternal diet supplemented with docosahexaenoic acid (DHA) during the suckling period on the development of the immune system and oral tolerance (OT) in offspring. Dams were randomized to consume one of two nutritionally adequate diets throughout the suckling period: control (N = 12, 0% DHA) or DHA (N = 8, 0.9% DHA) diet. At 11 days, pups from each dam were randomly assigned to a mucosal OT challenge: the placebo or the ovalbumin (OVA) treatment. At three weeks, plasma immunoglobulins and splenocyte cytokine production ex vivo were measured. OVA-tolerized pups had a lower Th2 (IL-13) response to OVA despite the presence of more activated T cells and memory cells (CD27+, all p < 0.05). Feeding a high DHA diet improved the ability of splenocytes to respond to mitogens toward a skewed Th1 response and led to a higher IL-10 and a lower TGF-β production after stimulation with OVA (all p < 0.05). Untolerized DHA-fed pups had lower plasma concentrations of OVA-specific immunoglobulin E (p for interaction < 0.05). Overall, feeding a high DHA maternal diet improves the tolerance response in untolerized suckled pups in a direction that is thought to be beneficial for the establishment of OT.

Topics: Animals; Dietary Supplements; Docosahexaenoic Acids; Egg Proteins; Female; Food Hypersensitivity; Immune Tolerance; Immunoglobulin E; Interleukin-10; Lactation; Maternal Nutritional Physiological Phenomena; Ovalbumin; Pregnancy; Rats, Sprague-Dawley; Spleen; T-Lymphocytes; Th1-Th2 Balance; Transforming Growth Factor beta

The number of patients with food allergy (FA) has dramatically increased. Although satisfactory drug therapies for FA are not available, we have found that kakkonto, a traditional Japanese herbal medicine, suppressed the occurrence of allergic symptoms in an FA mouse model. Thus, we investigated whether kakkonto could regulate the activation and differentiation of T cells in the colon.. BALB/c mice were systemically sensitized and then orally challenged with ovalbumin. FA mice were orally treated with kakkonto. Lamina propria (LP) cells from their colons were isolated and analyzed.. Kakkonto significantly reduced the proportion of CD69+ cells and the elevated helper T cell type 2-specific transcription factor GATA-3 mRNA expression in the LP CD4+ T cells, showing that kakkonto has a suppressive effect on the activation and Th2 differentiation of LP effector CD4+ T cells of the FA mouse colon. Furthermore, kakkonto significantly increased the proportion of Foxp3+CD4+ regulatory T cells in the LP CD4+ T cells of the FA mouse colon. Similarly, the number of Foxp3-positive cells was dramatically increased in the colonic mucosa of kakkonto-administered FA mice. However, the pharmacological effect and Foxp3+CD4+ regulatory T cell-inducing ability of kakkonto were not attenuated by the administration of an anti-CD25 monoclonal antibody in the FA model.. The induction of Foxp3+CD4+CD25- regulatory T cells in the colon as a novel mechanism underlying the therapeutic action of kakkonto could be utilized for the development of a novel anti-FA drug.

Topics: Animals; Cytokines; Disease Models, Animal; Drugs, Chinese Herbal; Food Hypersensitivity; Herbal Medicine; Immunophenotyping; Intestinal Mucosa; Male; Mice; Ovalbumin; Phenotype; T-Lymphocytes, Regulatory; Th2 Cells

The rapid increase in atopic diseases is potentially linked to increased hapten exposure, however, the role of haptens in the pathogenesis of food allergy remains unknown. Further studies are required to elucidate the cluster of differentiation 4 positive (CD4+) T cell response to food allergy induced by haptens. Dendritic cells were primed by trinitrobenzene sulfonic acid (TNBS) as a hapten or ovalbumin (OVA) as a model antigen, in a cell culture model. BALB/c mice were sensitized using TNBS and/or OVA. Intestinal Th1/Th2 cell and ovalbumin specific CD4+ T cells proliferation, intestinal cytokines (interleukin‑4 and interferon‑γ) in CD4+ T cells were evaluated. TNBS increased the expression of T cell immunoglobulin and mucin domain‑4 and tumor necrosis factor ligand superfamily member 4 in dendritic cells. Skewed Th2 cell polarization, extensive expression of interleukin‑4, reduced expression of interferon‑γ and forkhead box protein P3 were elicited following concomitant exposure to TNBS and OVA, with reduced regulatory T cells in the mouse intestinal mucosa, whereas a Th1 response was detected when challenged by TNBS or OVA alone. This data suggests that TNBS, as a hapten, combined with food antigens may lead to a Th2 cell response in the intestinal mucosa.

Topics: Animals; Food Hypersensitivity; Immunity, Mucosal; Interferon-gamma; Interleukin-4; Intestinal Mucosa; Male; Mice; Mice, Inbred BALB C; Ovalbumin; Th1 Cells; Th2 Cells; Trinitrobenzenesulfonic Acid

Although the incidence of food allergy continues to rise, there have been no effective therapeutic strategies. Citrus fruits contain a number of bioactive flavonoids with immune-regulatory functions. The objective of this study was to determine whether Citrus tachibana (fruit body with peel, leaves, and branch) can protect against the development of food allergy and the mechanism behind it, and to identify the active compound(s) responsible. We found that C. tachibana leaf extract (CLE) mitigated ovalbumin (OVA)-induced food allergy symptoms including increased rectal temperature, diarrhea, and anaphylaxis. This mitigation was likely due to CLE-mediated decreases in cytokine release from T-helper 2 cells (Th2 cells) in mesenteric lymph nodes. Moreover, higher levels of CLE attenuated systemic Th2 cell-mediated responses in mouse splenocytes sensitized with OVA+Alum. This was evidenced by CLE-mediated reductions in Th2 cytokine release, including interleukin (IL)-4, IL-5, and IL-13, but not the Th1 cytokines IL-12 and interferon (IFN)-γ, which was attributable to decreased gene expression levels. We also identified kaempferol as the most potent compound for reducing Th2-associated responses in splenocytes. The findings of this study suggest that CLE suppresses Th2-cell-mediated immune responses, contributing to alleviation of food allergy symptoms, and that kaempferol is a flavonoid with potential antiallergenic activity that targets Th2 cell-induced responses.

Topics: Alum Compounds; Animals; Cell Survival; Cells, Cultured; Citrus; Cytokines; Female; Food Hypersensitivity; Fruit; Kaempferols; Mice; Mice, Inbred BALB C; Ovalbumin; Plant Extracts; Plant Leaves; Spleen; Th2 Cells

Ovalbumin (OA) is the most abundant ingredient of chicken egg-white allergenic proteins. In the present study we investigated the possibility of reducing OA allergenicity by treatment with a natural protein exhibiting N-acetylglucosaminidase (NA) activity. Ascidian is cultivated as a food resource in northeast Asia. The ascidian viscera NA (AVNA) with almost no other exoglycosidases or proteolytic enzymes was isolated by applying size-exclusion chromatography to a protein precipitate of ascidian viscera. Intact OA was mixed with AVNA containing 0.2, 1.0, and 5.0 Units of NA. Anion-exchange chromatography was then used to isolate OA from AVNA-treated OA. The electrophoretic patterns and N-glycans of each isolated OA from AVNA-treated OA (iOA) were analyzed, and the terminal N-acetylglucosamines of iOA were selectively cleaved with no other degradation occurring. A competitive indirect enzyme-linked immunosorbent assay using rabbit anti-OA sera was performed to investigate the allergenicity of iOA, which was found to be significantly reduced depending on the increased NA activity compared to that of intact OA. These results indicate that OA allergenicity was reduced using a simple and mild treatment process with AVNA, and suggest that ascidian NA is an efficient natural protein for reducing the allergenicity of OA without requiring the use of harsh physical treatments or chemical conjugation.

Topics: Acetylglucosaminidase; Allergens; Animals; Chickens; Egg White; Food Hypersensitivity; Ovalbumin; Rabbits; Urochordata; Viscera

IL-10 is a key pleiotropic cytokine that can both promote and curb Th2-dependent allergic responses. In this study, we demonstrate a novel role for IL-10 in promoting mast cell expansion and the development of IgE-mediated food allergy. Oral OVA challenge in sensitized BALB/c mice resulted in a robust intestinal mast cell response accompanied by allergic diarrhea, mast cell activation, and a predominance of Th2 cytokines, including enhanced IL-10 expression. In contrast, the development of intestinal anaphylaxis, including diarrhea, mast cell activation, and Th2 cytokine production, was significantly attenuated in IL-10(-/-) mice compared with wild-type (WT) controls. IL-10 also directly promoted the expansion, survival, and activation of mast cells; increased FcεRI expression on mast cells; and enhanced the production of mast cell cytokines. IL-10(-/-) mast cells had reduced functional capacity, which could be restored by exogenous IL-10. Similarly, attenuated passive anaphylaxis in IL-10(-/-) mice could be restored by IL-10 administration. The adoptive transfer of WT mast cells restored allergic symptoms in IL-10(-/-) mice, suggesting that the attenuated phenotype observed in these animals is due to a deficiency in IL-10-responding mast cells. Lastly, transfer of WT CD4 T cells also restored allergic diarrhea and intestinal mast cell numbers in IL-10(-/-) mice, suggesting that the regulation of IL-10-mediated intestinal mast cell expansion is T cell dependent. Our observations demonstrate a critical role for IL-10 in driving mucosal mast cell expansion and activation, suggesting that, in its absence, mast cell function is impaired, leading to attenuated food allergy symptoms.

Topics: Adoptive Transfer; Animals; Cytokines; Diarrhea; Food Hypersensitivity; Immunoglobulin E; Interleukin-10; Intestines; Lymphocyte Activation; Mast Cells; Mice; Mice, Inbred BALB C; Ovalbumin; Receptors, IgE; Th2 Cells

Regulatory T cells (Tregs) are pivotal for maintenance of immune self-tolerance and also regulate immune responses to exogenous Ags, including allergens. Both decreased Treg number and function have been reported in allergic patients, offering new therapeutic perspectives. We previously demonstrated that Tregs can be selectively expanded and activated by low doses of IL-2 (ld-IL-2) inducing immunoregulation without immunosuppression and established its protective effect in autoimmune diseases. In this study, we evaluated the ability of ld-IL-2 to control allergy in an experimental model of food allergy. Ld-IL-2 induced Treg expansion and activation that elicited protection against clinical manifestations of food allergy in two mouse models with OVA and peanut. This clinical effect was lost in Treg-depleted mice, demonstrating the major contribution of Tregs in ld-IL-2 efficacy. Mechanistic studies further indicated that protection from allergy could be explained by a Treg-dependent local modification of the Th1/Th2 balance and an inhibition of mast cell recruitment and activation. Preventive and therapeutic effects of ld-IL-2 were observed over a 7-mo-period, highlighting its long-term efficacy. This study demonstrated that ld-IL-2 is efficient to prevent and to treat allergic immune responses, and thus represents a promising therapeutic strategy for managing allergic diseases.

Topics: Allergens; Animals; Arachis; Cell Proliferation; Cells, Cultured; Disease Models, Animal; Female; Food Hypersensitivity; Humans; Immunotherapy; Interleukin-2; Mast Cells; Mice; Mice, Inbred BALB C; Ovalbumin; Self Tolerance; T-Lymphocytes, Regulatory; Th1-Th2 Balance

Platelet-activating factor (PAF) is known to be an important mediator of anaphylaxis. However, there is a lack of information in the literature about the role of PAF in food allergy. The aim of this work was to elucidate the participation of PAF during food allergy development and the consequent adipose tissue inflammation along with its alterations. Our data demonstrated that, both before oral challenge and after 7 days receiving ovalbumin (OVA) diet, OVA-sensitized mice lacking the PAF receptor (PAFR) showed a decreased level of anti-OVA IgE associated with attenuated allergic markers in comparison to wild type (WT) mice. Moreover, there was less body weight and adipose tissue loss in PAFR-deficient mice. However, some features of inflamed adipose tissue presented by sensitized PAFR-deficient and WT mice after oral challenge were similar, such as a higher rate of rolling leukocytes in this tissue and lower circulating levels of adipokines (resistin and adiponectin) in comparison to nonsensitized mice. Therefore, PAF signaling through PAFR is important for the allergic response to OVA but not for the adipokine alterations caused by this inflammatory process. Our work clarifies some effects of PAF during food allergy along with its role on the metabolic consequences of this inflammatory process.

Topics: Adipokines; Adipose Tissue; Animal Feed; Animals; Biomarkers; Body Weight; Diet; Food Hypersensitivity; Immunoglobulin E; Inflammation; Leukocytes; Male; Mice; Mice, Inbred BALB C; Models, Animal; Ovalbumin; Platelet Membrane Glycoproteins; Receptors, G-Protein-Coupled

Since the early 2000s, labeling of potentially allergenic food components to protect people who suffer from food allergies is compulsory in numerous industrialized countries. In Europe, milk and egg components used during the winemaking process must be indicated on the label since July 1, 2012. Several ELISA procedures have been developed to detect allergenic residues in wines. However, the complexity of the wine matrix can inhibit the immunoenzymatic reaction. The aim of this study was to implement an ELISA assay for the detection of ovalbumin in red wines using commercially available antibodies. The specificity of the acquired antibodies and the absence of cross reactivity were assessed by immunoblotting and ELISA. An ELISA assay with a LOD of 14.2 μg/L and a LOQ of 56.4 μg/L of ovalbumin in aqueous solution was obtained. Differences in ELISA signals were observed when analyzing various fining agents, although reproducible conformation of the antigen could be reached for the comparison of ovalbumin and Ovicolle. The differences between samples in terms of pH could be leveled but the inhibition of the ELISA signal, positively correlated to the tannin content of the wines, could not be suppressed. Thus, standard curves of ovalbumin in several wines were obtained by relative quantification. The control steps and the difficulties encountered presented in this study should be considered by anybody working toward the development of ELISA assays for the detection of allergenic residues in complex food matrices.

Topics: Allergens; Animals; Antibodies; Cross Reactions; Eggs; Enzyme-Linked Immunosorbent Assay; Europe; Food Contamination; Food Hypersensitivity; Humans; Milk; Ovalbumin; Tannins; Wine

Cutaneous exposure to food allergens predisposes to food allergy, which is commonly associated with atopic dermatitis (AD). Levels of the epithelial cytokine IL-33 are increased in skin lesions and serum of patients with AD. Mast cells (MCs) play a critical role in food-induced anaphylaxis and express the IL-33 receptor ST2. The role of IL-33 in patients with MC-dependent food anaphylaxis is unknown.. We sought to determine the role and mechanism of action of IL-33 in patients with food-induced anaphylaxis in a model of IgE-dependent food anaphylaxis elicited by oral challenge of epicutaneously sensitized mice.. Wild-type, ST2-deficient, and MC-deficient Kit. Il33 mRNA expression was upregulated in tape-stripped mouse skin and scratched human skin. Tape stripping caused local and systemic IL-33 release in mice. ST2 deficiency, as well as ST2 blockade before oral challenge, significantly reduced the severity of oral anaphylaxis without affecting the systemic T. IL-33 is released after mechanical skin injury, enhances IgE-mediated MC degranulation, and promotes oral anaphylaxis after epicutaneous sensitization by targeting MCs. IL-33 neutralization might be useful in treating food-induced anaphylaxis in patients with AD.

Topics: Administration, Cutaneous; Allergens; Anaphylaxis; Animals; Dermatitis, Atopic; Female; Food Hypersensitivity; Humans; Immunoglobulin E; Interleukin-33; Mast Cells; Mice, Inbred BALB C; Mice, Transgenic; Ovalbumin; RNA, Messenger; Skin

To establish a food allergy model in Brown Norway (BN) rats by gavage of ovalbumin (OVA) without any adjuvant, and to evaluate this model.. A total of 20 male BN rats aged 3 weeks were randomly divided into allergy group and control group (n=10 each). BN rats in the allergy group were given OVA 1 mg per day by gavage, and all the rats were treated for 41 days continuously. On day 42, the rats in the allergy group were given OVA 100 mg by gavage for challenge. The rats in the control group were given normal saline of the same volume by gavage. Differences in body length, body weight, and food intake were compared between the two groups on days 7, 14, 21, 28, 35, and 42. ELISA was used to measure the serum OVA-IgE level and plasma histamine level after challenge on day 42, and the changes in rats' appearance and fecal properties were observed. The model of food allergy was considered successful when the serum OVA-IgE level in the allergy group was no less than the mean serum OVA-IgE level + 3 standard deviation in the control group.. There were no significant differences in body length, body weight or food intake between the allergy and control groups at all time points (P>0.05). On day 21, the control group had a significantly higher food intake than the allergy group (P<0.05). On day 42 after challenge, the allergy group showed significantly higher serum OVA-IgE and plasma histamine levels than the control group (P<0.05). The sensitization rate (rate of successful modeling) was 90%. The fecal properties showed no significant differences between the two groups.. OVA by gavage without any adjuvant can successfully establish the model of food allergy in BN rats and has a high success rate. Food allergy induced by OVA may reduce food intake within a short period of time, but no influence on rats' body length or body weight has been observed.

Topics: Animals; Disease Models, Animal; Food Hypersensitivity; Histamine; Immunoglobulin E; Male; Ovalbumin; Rats; Rats, Inbred BN

Humans could become exposed to carbon black nanoparticles (CBNPs) in consumer products or an occupational setting. In rodent models, acute respiratory, subcutaneous, and direct immune cell exposure to CBNPs has been shown to enhance allergic sensitization to co-administered ovalbumin (OVA) protein from chicken egg. However, little is known about the effects of ingested CBNPs on immunological responses and oral tolerance to food antigens. We hypothesized that ingestion of CBNPs would enhance the development of food allergy to OVA. Allergy prone DO11.10 mice were orally exposed to CBNPs every second day for 2 weeks (total dose 10.8 (LOW) or 108 μg (HI)), with and without (±) co-administered OVA. Systemic immune parameters were measured at necropsy. Exposure to OVA resulted in significant increases in serum anti-OVA IgG1, anti-OVA IgM, and anti-OVA IgA antibodies relative to vehicle control. Immunophenotyping revealed a reduction in the number of OVA-specific CD4

Topics: Administration, Oral; Animals; Cytokines; Female; Food Hypersensitivity; Immunoglobulins; Male; Mice; Mice, Inbred BALB C; Mice, Transgenic; Nanoparticles; Ovalbumin; Receptors, Antigen, T-Cell; Soot; Spleen; T-Lymphocytes, Helper-Inducer; Th2 Cells

The incidence of food allergy, which is triggered by allergen permeation of the gastrointestinal tract followed by a T-helper (Th) 2-mediated immune response, has been increasing annually worldwide. We examined the effects of baicalein (5,6,7-trihydroxyflavone), a flavonoid from Scutellaria baicalensis used in oriental herbal medicine, on regulatory T (Treg) cell induction and intestinal barrier function through the regulation of tight junctions in a mouse model of food allergy. An allergic response was induced by oral challenge with ovalbumin, and the incidence of allergic symptoms and T cell-related activity in the mesenteric lymph nodes were analyzed with and without the presence of baicalein. Our results demonstrated that the administration of baicalein ameliorated the symptoms of food allergy and attenuated serum IgE and effector T cells. However, Treg-related factors were up-regulated by baicalein. Furthermore, baicalein was shown to enhance intestinal barrier function through the regulation of tight junctions. We also found that baicalein treatment induced the differentiation of Treg cells via aryl hydrocarbon receptors (AhRs). Thus, the action of baicalein as an agonist of AhR can induce Treg differentiation and enhance barrier function, suggesting that baicalein might serve as an effective immune regulator derived from foods for the treatment of food allergy.

Topics: Animals; Disease Models, Animal; Female; Flavanones; Food Hypersensitivity; Forkhead Transcription Factors; Intestinal Mucosa; Intestines; Mice, Inbred BALB C; Ovalbumin; Phytotherapy; Plant Extracts; Receptors, Aryl Hydrocarbon; Scutellaria baicalensis; T-Lymphocytes, Regulatory; Tight Junctions

The pathogenesis of intestinal chronic inflammation is unclear. Food allergy plays an important role in the induction of intestinal inflammation. This study aims to test a hypothesis that food allergy initiates colitis. In this study, BALB/c mice were sensitized to a common food allergen, ovalbumin (OVA) with cholera toxin (CT) as an adjuvant. The colon epithelial barrier function was assessed with Ussing chamber technique. Expression of T cell immunoglobulin mucin domain molecule-4 (TIM4) in dendritic cells was evaluated by flow cytometry, RT-PCR and Western blotting. The results showed that allergen-related colitis was induced in mice as shown by heavy infiltration of inflammatory cells in the colon mucosa, loss of body weight of mice, increases in myeloperoxidase, tumor necrosis factor-α, interleukin-4, OVA-specific IgE in the colon tissue. The colon epithelial barrier function was markedly compromised in colitis group mice, which was mimicked by exposure the colon mucosa to CT in Ussing chamber. High frequency of TIM4(+) dendritic cells was detected in the colon mucosa of colitis mice. Exposure of dendritic cells to CT markedly increased the expression of TIM4. We conclude that IBD-like inflammation can be induced in the mouse colon by the food allergen-related immune response.

Topics: Animals; Cholera Toxin; Colitis; Colon; Dendritic Cells; Disease Models, Animal; Food Hypersensitivity; Inflammatory Bowel Diseases; Intestinal Mucosa; Membrane Proteins; Mice, Inbred BALB C; Ovalbumin; Th2 Cells

The early postnatal period is critical for immunity, and feeding docosahexaenoic acid (DHA) has been demonstrated to affect immune development.. The objective of this study was to determine the importance of feeding DHA during suckling and/or weaning on immune function and oral tolerance (OT).. Sprague-Dawley rats were randomly assigned to 1 of 2 nutritionally adequate diets throughout lactation (21 d): a control (n = 12, 0% DHA) diet or a DHA (n = 8, 0.9% DHA) diet. At 11 d, suckled pups from each dam were randomly assigned to a mucosal OT challenge: placebo or ovalbumin. At week 5, all pups systemically received ovalbumin + adjuvant to induce systemic immunization. At 21 d, pups from each dam were randomly assigned to 1 of the 2 diets for 21 d in a factorial design after which immune function and OT were assessed.. Feeding dams DHA during lactation resulted in a 40-60% higher splenocyte production of interleukin (IL)-10 when stimulated with concanavalin A, lipopolysaccharide (LPS), or ovalbumin and a 100% higher production of interferon (IFN)-γ with LPS (P < 0.05) than feeding the control diet to the pups. In comparison with pups fed the control diet, feeding DHA at weaning resulted in a 25% lower type 1 T helper (IL-1β) and type 2 T helper (IL-6) response by splenocytes after LPS stimulation and a 33% lower plasma concentration of ovalbumin-specific immunoglobulin (Ig) G (P < 0.05). Pups that did not receive additional DHA during the study had a 70% higher plasma concentration of ovalbumin-specific IgE than did the pups that received DHA at suckling and/or weaning (P < 0.05).. Feeding additional DHA during suckling had a beneficial programming effect on the ability of immune cells to produce IFN-γ and IL-10, and feeding DHA during weaning resulted in a lower inflammatory response. Providing no dietary DHA in either of the critical periods of immune development prevented the establishment of OT in female rat offspring.

Topics: Aging; Animal Nutritional Physiological Phenomena; Animals; Animals, Suckling; Cytokines; Dietary Supplements; Docosahexaenoic Acids; Female; Food Hypersensitivity; Immune Tolerance; Immunoglobulin G; Lactation; Maternal Nutritional Physiological Phenomena; Mitogens; Ovalbumin; Rats; Rats, Sprague-Dawley; Weaning

In our mouse model, gastric acid-suppression is associated with antigen-specific IgE and anaphylaxis development. We repeatedly observed non-responder animals protected from food allergy. Here, we aimed to analyse reasons for this protection. Ten out of 64 mice, subjected to oral ovalbumin (OVA) immunizations under gastric acid-suppression, were non-responders without OVA-specific IgE or IgG1 elevation, indicating protection from allergy. In these non-responders, allergen challenges confirmed reduced antigen uptake and lack of anaphylactic symptoms, while in allergic mice high levels of mouse mast-cell protease-1 and a body temperature reduction, indicative for anaphylaxis, were determined. Upon OVA stimulation, significantly lower IL-4, IL-5, IL-10 and IL-13 levels were detected in non-responders, while IL-22 was significantly higher. Comparison of fecal microbiota revealed differences of bacterial communities on single bacterial Operational-Taxonomic-Unit level between the groups, indicating protection from food allergy being associated with a distinct microbiota composition in a non-responding phenotype in this mouse model.

Topics: Administration, Oral; Allergens; Anaphylaxis; Animals; Anti-Ulcer Agents; Bacteria; Cytokines; Disease Models, Animal; Feces; Female; Food Hypersensitivity; Gastric Acid; Immunization; Immunoglobulin A; Immunoglobulin E; Immunoglobulin G; Intestines; Mice, Inbred BALB C; Microbiota; Ovalbumin; Spleen; Stomach; Sucralfate

To support dietary management of severe cow's milk allergic infants, a synbiotic mixture of non-digestible oligosaccharides and

Topics: Animals; Bifidobacterium breve; Cattle; Chickens; Disease Models, Animal; Female; Food Hypersensitivity; Humans; Infant; Mice; Mice, Inbred BALB C; Oligosaccharides; Ovalbumin; Synbiotics

The ability to avoid inflammatory responses to dietary components and microbiota antigens in the gut mucosa is achieved by a mechanism termed oral tolerance. This phenomenon is crucial to maintain the physiological immune activity in the gut and to prevent inflammatory disorders such as food allergy and inflammatory bowel diseases. Moreover, orally administered antigens induce regulatory cells that control systemic inflammatory responses as well. Given its specific, systemic and long-lasting effects, oral tolerance represents a promising approach for immunotherapies that aim to modulate inflammatory and autoimmune diseases. However, there are different protocols of feeding for induction of oral tolerance, and they have an impact in tolerance efficiency and length. Herein, we present and discuss different experimental feeding protocols and how they influence the outcome of oral administration of antigens.

Topics: Administration, Oral; Animals; Desensitization, Immunologic; Enteral Nutrition; Female; Food Hypersensitivity; Immune Tolerance; Immunoglobulin E; Inflammatory Bowel Diseases; Intestinal Mucosa; Male; Mice; Mice, Inbred BALB C; Mice, Inbred C57BL; Ovalbumin; Th1 Cells

Food allergy (FA) is an adverse health effect produced by the exposure to a given food. Currently, there is no optimal animal model of FA for the screening of immunotherapies or for testing the allergenicity of new foods.. The aim of the present study was to develop an effective and rapid model of FA in Brown Norway rats. In order to establish biomarkers of FA in rat, we compared the immune response and the anaphylactic shock obtained in this model with those achieved with only intraperitoneal immunization.. Rats received an intraperitoneal injection of ovalbumin (OVA) with alum and toxin from Bordetella pertussis, and 14 days later, OVA by oral route daily for three weeks (FA group). A group of rats receiving only the i.p. injection (IP group) were also tested. Serum anti-OVA IgE, IgG1, IgG2a, IgG2b and IgA antibodies were quantified throughout the study. After an oral challenge, body temperature, intestinal permeability, motor activity, and mast cell protease II (RMCP-II) levels were determined. At the end of the study, anti-OVA intestinal IgA, spleen cytokine production, lymphocyte composition of Peyer's patches and mesenteric lymph nodes, and gene expression in the small intestine were quantified.. Serum OVA-specific IgG1, IgG2a and IgG2b concentrations rose with the i.p. immunization but were highly augmented after the oral OVA administration. Anti-OVA IgE increased twofold during the first week of oral OVA gavage. The anaphylaxis in both IP and FA groups decreased body temperature and motor activity, whereas intestinal permeability increased. Interestingly, the FA group showed a much higher RMCP II serum protein and intestinal mRNA expression.. These results show both an effective and relatively rapid model of FA assessed by means of specific antibody titres and the high production of RMCP-II and its intestinal gene expression.

Topics: Animals; Biomarkers; Bordetella pertussis; Disease Models, Animal; Food Hypersensitivity; Gene Expression Regulation; Humans; Immunoglobulin A; Immunoglobulin E; Immunoglobulin G; Intestines; Ovalbumin; Rats; Serine Endopeptidases

Screen-printed platinum electrodes as transducer and magnetic beads as solid phase were combined to develop a particle-based electrochemical immunosensor for monitoring the serious food allergen ovalbumin. The standard arrangement of enzyme-linked immunosorbent assay became the basis for designing the immunosensor. A sandwich-type immunocomplex was formed between magnetic particles functionalized with specific anti-ovalbumin immunoglobulin G and captured ovalbumin molecules, and secondary anti-ovalbumin antibodies conjugated with the enzyme horseradish peroxidase were subsequently added as label tag. The electrochemical signal proportional to the enzymatic reaction of horseradish peroxidase during the reduction of hydrogen peroxide with thionine as electron mediator was measured by linear sweep voltammetry. The newly established method of ovalbumin detection exhibits high sensitivity suitable for quantification in the range of 11 to 222nM and a detection limit of 5nM. Magnetic beads-based assay format using external magnets for rapid and simple separation has been proven to be an excellent basis for electrochemical detection and quantification of food allergens in highly complex sample matrices.

Topics: Animals; Antibodies, Immobilized; Biosensing Techniques; Electric Conductivity; Electrochemistry; Electrodes; Electron Transport; Food Hypersensitivity; Immunoassay; Limit of Detection; Magnets; Microspheres; Ovalbumin; Platinum

IgE antibodies and mast cells play critical roles in the establishment of allergic responses to food antigens. Curcumin, the active ingredient of the curry spice turmeric, has anti-inflammatory properties, and thus may have the capacity to regulate Th2 cells and mucosal mast cell function during allergic responses. We assessed whether curcumin ingestion during oral allergen exposure can modulate the development of food allergy using a murine model of ovalbumin (OVA)-induced intestinal anaphylaxis. Herein, we demonstrate that frequent ingestion of curcumin during oral OVA exposure inhibits the development of mastocytosis and intestinal anaphylaxis in OVA-challenged allergic mice. Intragastric (i.g.) exposure to OVA in sensitized BALB/c mice induced a robust IgE-mediated response accompanied by enhanced OVA-IgE levels, intestinal mastocytosis, elevated serum mMCP-1, and acute diarrhea. In contrast, mice exposed to oral curcumin throughout the experimental regimen appeared to be normal and did not exhibit intense allergic diarrhea or a significant enhancement of OVA-IgE and intestinal mast cell expansion and activation. Furthermore, allergic diarrhea, mast cell activation and expansion, and Th2 responses were also suppressed in mice exposed to curcumin during the OVA-challenge phase alone, despite the presence of elevated levels of OVA-IgE, suggesting that curcumin may have a direct suppressive effect on intestinal mast cell activation and reverse food allergy symptoms in allergen-sensitized individuals. This was confirmed by observations that curcumin attenuated the expansion of both adoptively transferred bone marrow-derived mast cells (BMMCs), and inhibited their survival and activation during cell culture. Finally, the suppression of intestinal anaphylaxis by curcumin was directly linked with the inhibition of NF-κB activation in curcumin-treated allergic mice, and curcumin inhibited the phosphorylation of the p65 subunit of NF-κB in BMMCs. In summary, our data demonstrates a protective role for curcumin during allergic responses to food antigens, suggesting that frequent ingestion of this spice may modulate the outcome of disease in susceptible individuals.

Topics: Anaphylaxis; Animals; Curcumin; Disease Models, Animal; Food Hypersensitivity; Intestinal Mucosa; Intestines; Mast Cells; Mastocytosis; Mice; NF-kappa B; Ovalbumin; Phosphorylation; Signal Transduction

Prostaglandin D2 (PGD2) is a major prostanoid secreted mainly by mast cells. Although PGD2 has been identified as a modulator of allergic inflammation, its precise role remains unclear. Here we investigate the role of PGD2 in food allergy. Oral administration of ovalbumin induces allergic responses in sensitized wild-type (WT) mice. Systemic gene deficiency of haematopoietic PGD synthase (H-PGDS(-/-)) exacerbates all of the manifestations accompanying severe mast cell hyperplasia in the intestine. Morphological studies show that c-kit/FcɛRI-positive WT mast cells strongly express H-PGDS. Transplantation of H-PGDS(-/-) mast cells also aggravates ovalbumin-induced mast cell hyperplasia and allergic symptoms in mast cell null mice. H-PGDS deficiency accelerates the production of SDF-1α and the activity of MMP-9 in the antigen-stimulated intestine. SDF-1α receptor blockade or MMP-9 inhibition relieves the exacerbated mast cell hyperplasia and manifestations observed in H-PGDS(-/-). Thus, PGD2 deficiency results in food antigen-induced mast cell hyperplasia.

Topics: Adjuvants, Immunologic; Alum Compounds; Animals; Chemokine CXCL12; Colon; Cytokines; Enzyme-Linked Immunosorbent Assay; Food Hypersensitivity; Hyperplasia; Intestines; Intramolecular Oxidoreductases; Lipocalins; Mast Cells; Matrix Metalloproteinase 9; Mice; Mice, Inbred C57BL; Mice, Knockout; Microscopy, Electron; Ovalbumin; Prostaglandin D2; Reverse Transcriptase Polymerase Chain Reaction

Toll-like receptor 2 (TLR2) is a widely expressed pattern recognition receptor critical for innate immunity. TLR2 is also a key regulator of mucosal immunity implicated in the development of allergic disease. TLR2 activators are found in many common foods, but the role of TLR2 in oral tolerance and allergic sensitization to foods is not well understood.. The purpose of this study was to evaluate the impacts of TLR2 expression and TLR2 activation on oral tolerance to food antigens in a murine model.. Mice were fed ovalbumin (OVA) or peanut butter with or without the addition of low doses of TLR2 activators Pam3 CSK4 or FSL-1. Oral tolerance was assessed by analysing antibody responses after a systemic antigen challenge. OVA-specific Tregs were assessed in the Peyer's patches, mesenteric lymph nodes, and spleen in wild-type and TLR2(-/-) mice. Low-dose Pam3 CSK4 was also tested as an oral adjuvant.. Oral tolerance was successfully induced in both wild-type and TLR2(-/-) recipient mice, with an associated regulatory T-cell response. Oral TLR2 activation, with low-dose Pam3 CSK4 or FSL-1, during oral antigen exposure was found to alter oral tolerance and was associated with the development of substantial IgE and IgA responses to foods upon systemic challenge. Low-dose oral Pam3 CSK4 treatment also selectively enhanced antigen-specific IgA responses to oral antigen exposure.. TLR2 is not necessary for oral tolerance induction, but oral TLR2 activation modulates humoral IgE and IgA responses during tolerance development. Low-dose Pam3 CSK4 is also an effective oral adjuvant that selectively enhances IgA production. These observations are pertinent to the optimization of oral allergen immunotherapy and oral vaccine development.

Topics: Adjuvants, Immunologic; Allergens; Animals; Diglycerides; Disease Models, Animal; Food; Food Hypersensitivity; Immune Tolerance; Immunity, Humoral; Immunoglobulin A; Lipopeptides; Mice; Mice, Knockout; Oligopeptides; Ovalbumin; T-Lymphocyte Subsets; T-Lymphocytes, Regulatory; Toll-Like Receptor 2

Fining of wine with agents containing cow's milk or hen's egg white is a common and traditional procedure. In light of increasing food allergies all over the world, the presence of fining residues has been subject of intense debate. Switzerland does not make exception, and since 2009 the Federal Department of Home Affairs has modified its food regulations stating that the labels must show if traces of fining agents are present. Nevertheless, the application of this regulation is not based on an official analytical method. In this study we show that immunoblotting is an efficient technique to detect and quantify ovalbumin and casein residues in bottled wine. We showed that final filtration is an essential step to remove finings in red wine, and that overfining of white wine may result in fining residues in finished products. Finally, for the first time in Switzerland, 22 samples were taken by food safety inspectors and officially analyzed for the regional food control authority of the Canton of Vaud. These samples were allergen free, but a larger study is currently planned in collaboration with other regional authorities of Switzerland to complete these results and make a complete picture of the Swiss wine production.

Topics: Allergens; Animals; Blotting, Western; Caseins; Cattle; Chickens; Eggs; Female; Food Handling; Food Hypersensitivity; Food Labeling; Humans; Milk; Ovalbumin; Switzerland; Wine

Trachelospermi caulis is used widely as an herbal medicine in oriental countries to attenuate fever and pain. We wished to reveal the novel function of this herb and its active component on barrier function in intestinal epithelial cells. Monolayers of intestinal epithelial cells (Caco-2) were used to evaluate the transepithelial electrical resistance (TEER) and quantity of permeated ovalbumin (OVA) as indices of barrier function. T. caulis increased TEER values on cell monolayers and decreased OVA permeation across cell monolayers. To ascertain the active component of T. caulis, the extract was isolated to five fractions, and the effect of each of these fractions on intestinal barrier function examined. Chloroform and ethyl acetate fractions showed increased TEER values and decreased OVA flux. Chloroform and ethyl acetate fractions contained mainly trachelogenin and its glycoside, tracheloside. Trachelogenin increased TEER values and decreased OVA flux by enhancing the tight-junction protein occludin (but not tracheloside) in Caco-2 monolayers. These findings demonstrated that trachelogenin, an active component of T. caulis, might help to attenuate food allergy or inflammatory bowel disease through inhibition of allergen permeation or enhancement of the intestinal barrier.

Topics: 4-Butyrolactone; Allergens; Apocynaceae; Caco-2 Cells; Colon; Food Hypersensitivity; Glucosides; Humans; Inflammatory Bowel Diseases; Intestinal Mucosa; Occludin; Ovalbumin; Permeability; Phytotherapy; Plant Extracts; Tight Junctions

To explore the effect of non-methylated cytosine-phosphate-guanine oligodeoxynucleotides (CpG-ODN) on serum transforming growth factor (TGF)-β and immune regulation in ovalbumin (OVA)-sensitized young mice.. Thirty female BALB/c mice (2-3 weeks old) were randomly divided into control, model, and CpG-ODN intervention groups. A young mouse model of food allergy was established by OVA sensitization. Normal saline of the same volume was used for replacement in the control group. The mice in the intervention group were intraperitoneally injected with CpG-ODN solution 1 hour before every OVA sensitization. Allergic symptoms were observed and scored for each group. The jejunal tissue was histopathologically examined with hematoxylin-eosin staining. Serum OVA-IgE level was measured using ELISA. Serum concentrations of interleukin (IL)-4, interferon (IFN)-γ, and TGF-β were determined by CBA.. Allergic symptoms were observed in the model group and the jejunal tissue showed the pathological characteristics of type I allergic reaction. The allergic symptom scores in the model and CpG-ODN intervention groups were significantly higher than in the control group (P<0.01). The serum levels of OVA-IgE, IL-4, and TGF-β were significantly higher in the model group than in the control and CpG-ODN intervention groups (P<0.05). The CpG-ODN intervention group had significantly higher serum levels of OVA-IgE, IL-4, and TGF-β than the control group (P<0.05). Compared with the control and CpG-ODN intervention groups, the model group had a significantly reduced IFN-γ level (P<0.05).. The serum TGF-β level is increased in the young mouse model of OVA-sensitized food allergy and is involved in the allergy mechanism. Non-methylated CpG-ODN can reduce the serum TGF-β level in sensitized young mice and play an immunoregulatory role in food allergy.

Topics: Aging; Animals; DNA Methylation; Female; Food Hypersensitivity; Immunoglobulin E; Interleukin-4; Mice; Mice, Inbred BALB C; Oligodeoxyribonucleotides; Ovalbumin; Transforming Growth Factor beta

The neonatal period is often polarised to T helper (Th2) response at the time of birth, predisposing offspring to allergic disorders. Passive immunity through the mother's milk is critical for immune system development of newborns. Probiotics have been proposed to harmonise Th1/Th2 imbalance in allergic conditions in adults. In the present study, the anti-allergic effects of feeding probiotic Lactobacillus rhamnosus-fermented milk (PFM) either to dams during the suckling period or to their offspring after weaning individually or else in successive periods against ovalbumin (OVA)-induced allergy in newborns was analysed. After allergen sensitisation, physical symptoms of allergy, gut immune response, humoral immune response and cell-mediated response through interleukins were detected. Consumption of PFM by mothers and offspring showed a reduction (P<0·01) in physical allergic symptoms in newborns with an increase (P<0·01) in the numbers of goblet and IgA+ cells in the small intestine. Similarly, considerable (P<0·001) decreases in OVA-specific antibodies (IgE, IgG, IgG1) and ratios of IgE/IgG2a and IgG1/IgG2a in the sera of newborn mice were recorded. A decrease in IL-4 and an increase in interferon-γ levels further confirmed the shift from Th2 to Th1 pathway in PFM-fed mice. It is logical to conclude that the timing of PFM intervention in alleviating allergic symptoms is critical, which was found to be most effective when mothers were fed during the suckling period.

Topics: Allergens; Animals; Anti-Allergic Agents; Chemokine CCL2; Cyclooxygenase 2; Female; Fermentation; Food Hypersensitivity; Immunoglobulin E; Immunoglobulin G; Interferon-gamma; Interleukin-10; Interleukin-4; Intestines; Lacticaseibacillus rhamnosus; Male; Mice; Milk; Ovalbumin; Probiotics; Th1 Cells; Th2 Cells; Toll-Like Receptor 2; Toll-Like Receptor 4; Transforming Growth Factor beta

Turmeric (Curcuma longa) has traditionally been used to treat pain, fever, allergic and inflammatory diseases such as bronchitis, arthritis, and dermatitis. In particular, turmeric and its active component, curcumin, were effective in ameliorating immune disorders including allergies. However, the effects of turmeric and curcumin have not yet been tested on food allergies.. Mice were immunized with intraperitoneal ovalbumin (OVA) and alum. The mice were orally challenged with 50mg OVA, and treated with turmeric extract (100mg/kg), curcumin (3mg/kg or 30 mg/kg) for 16 days. Food allergy symptoms including decreased rectal temperature, diarrhea, and anaphylaxis were evaluated. In addition, cytokines, immunoglobulins, and mouse mast cell protease-1 (mMCP-1) were evaluated using ELISA.. Turmeric significantly attenuated food allergy symptoms (decreased rectal temperature and anaphylactic response) induced by OVA, but curcumin showed weak improvement. Turmeric also inhibited IgE, IgG1, and mMCP-1 levels increased by OVA. Turmeric reduced type 2 helper cell (Th2)-related cytokines and enhanced a Th1-related cytokine. Turmeric ameliorated OVA-induced food allergy by maintaining Th1/Th2 balance. Furthermore, turmeric was confirmed anti-allergic effect through promoting Th1 responses on Th2-dominant immune responses in immunized mice.. Turmeric significantly ameliorated food allergic symptoms in a mouse model of food allergy. The turmeric as an anti-allergic agent showed immune regulatory effects through maintaining Th1/Th2 immune balance, whereas curcumin appeared immune suppressive effects. Therefore, we suggest that administration of turmeric including various components may be useful to ameliorate Th2-mediated allergic disorders such as food allergy, atopic dermatitis, and asthma.

Topics: Allergens; Anaphylaxis; Animals; Chymases; Curcuma; Curcumin; Cytokines; Disease Models, Animal; Female; Food Hypersensitivity; Immunoglobulin E; Immunoglobulin G; Mice, Inbred BALB C; Ovalbumin; Phytotherapy; Plant Extracts; Th1 Cells; Th2 Cells

The number of students with food allergy is currently increasing. Moreover, the unintentional mixing or accidental ingestion of allergy-causing food materials in school lunches has attracted great attention. The aim of this study was to verify the current status of elementary school lunch provision for students with food allergy. We investigated the elementary school lunch services in seven cities in Osaka prefecture. The egg elimination diet was provided in five of the seven cities. In four of these five cities, we did not detect the presence of egg residue either on the surface of various cookware used to prepare the egg elimination diet or in the food itself. In this investigation, the egg elimination diet was provided properly, but we observed differences among the cities in the manual preparation of foods for food allergy diets. To step up these efforts, our results suggest the necessity of preparing a manual to consider individual conditions of school lunch services.

Topics: Child; Cooking and Eating Utensils; Diet; Egg Hypersensitivity; Enzyme-Linked Immunosorbent Assay; Food Hypersensitivity; Food Services; Humans; Japan; Lunch; Ovalbumin; Schools

Experimental IgE-mediated food allergy depends on intestinal anaphylaxis driven by interleukin-9 (IL-9). However, the primary cellular source of IL-9 and the mechanisms underlying the susceptibility to food-induced intestinal anaphylaxis remain unclear. Herein, we have reported the identification of multifunctional IL-9-producing mucosal mast cells (MMC9s) that can secrete prodigious amounts of IL-9 and IL-13 in response to IL-33, and mast cell protease-1 (MCPt-1) in response to antigen and IgE complex crosslinking, respectively. Repeated intragastric antigen challenge induced MMC9 development that required T cells, IL-4, and STAT6 transcription factor, but not IL-9 signals. Mice ablated of MMC9 induction failed to develop intestinal mastocytosis, which resulted in decreased food allergy symptoms that could be restored by adoptively transferred MMC9s. Finally, atopic patients that developed food allergy displayed increased intestinal expression of Il9- and MC-specific transcripts. Thus, the induction of MMC9s is a pivotal step to acquire the susceptibility to IgE-mediated food allergy.

Topics: Adoptive Transfer; Anaphylaxis; Animals; Base Sequence; Bone Marrow Cells; Cell Lineage; Chymases; Diarrhea; Disease Susceptibility; Duodenum; Food Hypersensitivity; Humans; Hypersensitivity, Immediate; Immunoglobulin E; Interleukin-9; Interleukins; Intestinal Mucosa; Mast Cells; Mastocytosis; Mice; Mice, Inbred Strains; Molecular Sequence Data; Ovalbumin; RNA, Messenger; Species Specificity; STAT6 Transcription Factor; T-Lymphocytes

The mammalian circadian clock controls many physiological processes that include immune responses and allergic reactions. Several studies have investigated the circadian regulation of intestinal permeability and tight junctions known to be affected by cytokines. However, the contribution of circadian clock to food allergy symptoms remains unclear. Therefore, we investigated the role of the circadian clock in determining the severity of food allergies. We prepared an ovalbumin food allergy mouse model, and orally administered ovalbumin either late in the light or late in the dark period under light-dark cycle. The light period group showed higher allergic diarrhea and weight loss than the dark period group. The production of type 2 cytokines, IL-13 and IL-5, from the mesenteric lymph nodes and ovalbumin absorption was higher in the light period group than in the dark period group. Compared to the dark period group, the mRNA expression levels of the tight junction proteins were lower in the light period group. We have demonstrated that increased production of type 2 cytokines and intestinal permeability in the light period induced severe food allergy symptoms. Our results suggest that the time of food antigen intake might affect the determination of the severity of food allergy symptoms.

Topics: Allergens; Animals; Basic-Leucine Zipper Transcription Factors; Cell Membrane Permeability; Cytokines; Diarrhea; Disease Models, Animal; Food Hypersensitivity; Gene Expression Regulation; Immunoglobulin E; Intestinal Mucosa; Lymph Nodes; Mesentery; Mice; Occludin; Ovalbumin; Photoperiod; RNA, Messenger; Severity of Illness Index; Tight Junctions

Heated foods often present low allergenicity, and have recently been used in specific oral immunotherapy for food allergies. However, the influence of heating on tolerogenicity of food allergens is not well elucidated. Here, we investigated biochemical, allergenic, and tolerogenic properties of heated egg white (EW) using a murine model of food allergy.. Raw EWs were treated at 80°C for 15 min (80EW, mild heating condition), 100°C for 5 min (100EW, cooking condition), or 121°C for 40 min (121EW, retort pouch condition), and freeze-dried. A transgenic OVA23-3 mice model expressing T-cell receptor specific for ovalbumin (OVA, a major EW allergen) induced Th2 cells and IgE production, and presented intestinal inflammation when fed untreated EW diet. 80EW-fed mice presented only moderate inflammation but high Th2 responses. 100EW-fed mice did not present inflammation but induced tolerance as seen in reduced T-cell responses and IgE levels. 100EW demonstrated higher digestive stability and slower absorption in intestine, compared with untreated EW and 80EW. 121EW was strongly aggregated, was not absorbed well, and developed Th1 responses without tolerance induction.. OVA in EW treated only under a particular heat condition (e.g. 100°C for 5 min) lost allergenicity, but possessed tolerogenicity.

Topics: Animals; Cell Proliferation; Disease Models, Animal; Egg White; Food Handling; Food Hypersensitivity; Hot Temperature; Immune Tolerance; Immunoglobulin E; Inflammation; Intestines; Mice; Mice, Transgenic; Ovalbumin

To clarify the role of Janus kinase (JAK) in and the efficacy of JAK inhibitors on food allergy, we investigated the effect of the clinically available JAK inhibitor ruxolitinib on mouse food allergy and the functions of cultured mast cells in vitro. Anaphylactic symptoms including diarrhea and decreases in body temperature pursuant to oral ovalbumin (OVA) challenges in food allergy mice were attenuated by the daily oral administration of ruxolitinib. This drug inhibited increases in mouse mast cell protease-1 concentrations in the serum and mast cell numbers in the intestines of these mice as well as degranulation, IL-13 production, and the spontaneous and IL-9-dependent survival of mouse bone marrow-derived mast cells in spite of the absence of an effect of ruxolitinib on passive systemic anaphylaxis. Anti-OVA IgG2a, IgE, and IgG1 serum levels and the release of IFN-γ, IL-4, IL-9, and IL-10 from the OVA-restimulated splenocytes of food allergy mice were also decreased by the treatment. Moreover, ruxolitinib administration to mice that had already exhibited anaphylactic responses to previous challenges reduced anaphylactic responses to further oral OVA challenges, which suggested that ruxolitinib has a therapeutic potential on food allergy. Our results showed that ruxolitinib remitted food allergy in mice mainly through immunosuppression and the prevention of mast cell hyperplasia, and partially through the inhibition of mast cell activation. We consider JAK inhibition to be a promising strategy for the prevention of food allergy, and ruxolitinib along with its derivatives inhibiting JAK as good candidates for therapeutic drugs to treat food allergy.

Topics: Allergens; Animals; Bone Marrow Cells; Cell Line; Chymases; Cytokines; Food Hypersensitivity; Goblet Cells; Immunoglobulin E; Immunoglobulin G; Intestines; Janus Kinases; Male; Mast Cells; Mice; Mice, Inbred BALB C; Nitriles; Ovalbumin; Protein Kinase Inhibitors; Pyrazoles; Pyrimidines; Spleen

The Maillard reaction (also referred to as "glycation") takes place between reducing sugars and compounds with free amino groups during thermal processing of foods. In the final stage of the complex reaction cascade, the so-called advanced glycation end products (AGEs) are formed, including proteins with various glycation structures. It has been suggested that some AGEs could have immunostimulatory effects. Here, we aimed to identify specific glycation structure(s) that could influence the T-cell immunogenicity and potential allergenicity of food allergens, using ovalbumin (OVA, an egg white allergen) as a model allergen. OVA was specifically modified with representative glycation structures: N(ε)-carboxymethyl lysine (CM-OVA), N(ε)-carboxyethyl lysine (CE-OVA), pyrraline (Pyr-OVA), or methylglyoxal-derived arginine derivatives (MGO-OVA). As well as AGE-OVA, a crude glycation product in thermal incubation of OVA with glucose, only Pyr-OVA, and not other modified OVAs, was efficiently taken up by bone marrow-derived murine dendritic cells (BMDCs). The uptake of Pyr-OVA was reduced in scavenger receptor class A (SR-A)-deficient BMDCs, but not in cells treated with inhibitors of scavenger receptor class B, galectin-3, or blocking antibodies against CD36, suggesting that pyrraline binds to SR-A. Compared with other modified OVAs, Pyr-OVA induced higher activation of OVA-specific CD4(+) T-cells in co-culture with BMDCs. Furthermore, compared with native OVA, AGE-OVA and Pyr-OVA induced higher IgE production in mice. Pyrraline could induce better allergen uptake by DCs via association with SR-A and subsequently enhance CD4(+) T-cell activation and IgE production. Our findings help us to understand how Maillard reaction enhances the potential allergenicity of food allergens.

Topics: Allergens; Animals; Bone Marrow Cells; Carbohydrates; CD4-Positive T-Lymphocytes; Coculture Techniques; Cytokines; Dendritic Cells; Enzyme-Linked Immunosorbent Assay; Female; Food Hypersensitivity; Lymphocyte Activation; Maillard Reaction; Mice; Mice, Inbred BALB C; Mice, Inbred C57BL; Norleucine; Ovalbumin; Protein Structure, Secondary; Pyrroles; Receptors, Scavenger

Neonatal to early childhood is the critical period for establishing a balance of T helper 1 (Th1) versus T helper 2 (Th2) cellular immunity within the gut, which is strongly influenced by the source and establishment of gut microflora. Probiotic administration has been shown to attenuate Th2-biased cellular immunity and predisposition to food allergies. To test this hypothesis we provided ad libitum a probiotic-supplemented (Primalac 454 Feed Grade Microbials) or control diet to lactating dams with suckling pups and weaned pups until 10 weeks of age. Weaned mice were sensitized/challenged with egg allergen ovalbumin, saline or adjuvant at 6, 8 and 10 weeks of age. At 3, 6, 8 and 10 weeks, fecal samples were collected for microbial analysis, while blood samples were analyzed for ovalbumin-IgE and total plasma IgE levels. At termination, splenic T helper cell lymphocyte population subtypes were determined using FACS analysis and Th1/Th2/Th17 gene expression by PCR array. At 21 days of age, pups suckled by lactating dams fed the probiotic supplemented diet had significantly enhanced Lactobacillus acidophilus fecal counts compared to controls. Moreover, mice fed the probiotic supplemented diet had enhanced splenic naturally occurring and induced regulatory T cell populations, enhanced TGFβ gene expression and reduced expression of allergic mediator IL13 compared to controls. These results provide evidence that early probiotic supplementation may provide host protection from hypersensitivity reactions to food allergens by attenuating food allergen inflammatory responses.

Topics: Allergens; Animals; Animals, Newborn; Antibody Specificity; Body Weight; Dietary Supplements; Disease Models, Animal; Female; Food Hypersensitivity; Gastrointestinal Tract; Immunoglobulin E; Lactobacillus acidophilus; Maternal Exposure; Mice; Ovalbumin; Probiotics; Spleen; T-Lymphocytes, Helper-Inducer

Food allergy, which accompanies acute symptoms such as pruritus, vomiting, diarrhea, and lethal anaphylactic shock is an increasing clinical problem. Skullcap (Scutellaria baicalensis Georgi) has been widely used as a traditional herbal medicine to treat inflammation, cancer, and allergy, but its effects in treating food allergy are not yet known.. To examine the effect of skullcap on food allergy, female BALB/c mice were sensitized with 20 μg OVA and 2mg alum by intraperitoneal injection on day 0. From day 17, mice were orally challenged with OVA (50 mg) in saline every 3 days, for a total of six times. To investigate the preventive effect, skullcap (25 mg/kg) was orally administered every day from day 17 to 34.. Food allergy symptoms were evaluated by the criteria for diarrhea, anaphylactic response, and rectal temperature. Severe symptoms of food allergy were observed in the sham group (diarrhea, 3 points; anaphylactic response, 2.6 points; rectal temperature, -8.36 °C. In contrast, the skullcap treatment group had a significantly suppressed OVA-induced anaphylactic response (1.3 points) and rectal temperature (-4.76°C). Moreover, both OVA-specific IgE, Th17 cytokine (IL-17), and Th2-related cytokines (IL-4, IL-5, IL-10, and IL-13), which increased with food allergy, were significantly inhibited by skullcap treatment.. We demonstrate that the administration of skullcap attenuates OVA-induced food allergy symptoms through regulating systemic immune responses of Th cells. These results indicate that skullcap may be a potential candidate as a preventive agent for food allergy.

Topics: Allergens; Anaphylaxis; Animals; Anti-Allergic Agents; Body Temperature; Cytokines; Diarrhea; Disease Models, Animal; Female; Food Hypersensitivity; Immunoglobulin E; Immunoglobulin G; Mice, Inbred BALB C; Ovalbumin; Phytotherapy; Plant Extracts; Scutellaria baicalensis; Spleen

Polyphenols are naturally derived bioactive compounds with numerous reported health benefits. We have previously reported on the beneficial effect of a polyphenol-enriched apple extract in a murine model of food allergy. The objectives of the present study were to elucidate the class of bioactive polyphenols that exhibit a beneficial anti-allergic effect and to assess whether the protective effect matches the in vivo bioavailable metabolite concentrations. Female BALB/c mice were sensitised to ovalbumin (OVA) following the protocol of a well-established murine model of food allergy. They were fed diets containing polyphenol-enriched extracts or purified epicatechin for 8 d after the last sensitisation. The sensitised mice were orally challenged with OVA after the intervention. The allergy symptoms, in addition to allergen-specific serum Ig concentrations and gene expression profiles in the intestine, of the control and treated mice were compared. Plasma samples were collected to compare the concentrations of bioavailable epicatechin metabolites in the treatment groups. Polyphenol-enriched fruit extracts containing epicatechin exhibited a significant anti-allergic effect in vivo. This effect was unambiguously attributed to epicatechin, as oral administration of this purified polyphenol to sensitised mice by inclusion in their diet modulated allergy symptoms in a dose-dependent manner. Immune parameters were also affected by the administration of epicatechin. Bioavailability measurements in plasma indicated that the attenuation of allergy symptoms could be due to the higher concentrations of bioavailable epicatechin metabolites. In conclusion, epicatechin is a key bioactive polyphenol that has the ability to modulate allergy outcomes in sensitised mice.

Topics: Animals; Anti-Allergic Agents; Biological Availability; Catechin; Chymases; Cytokines; Disease Models, Animal; Female; Food Hypersensitivity; Fruit; Gene Expression; Immunoglobulin E; Immunoglobulin G; Intestine, Small; Lymph Nodes; Malus; Mesentery; Mice; Mice, Inbred BALB C; Ovalbumin; Plant Extracts; Polyphenols

Cutaneous sensitization with a food antigen before its consumption elicits the development of food allergy. Here, we report the site- and stage-dependent roles of basophils and proallergic cytokines, thymic stromal lymphopoietin (TSLP) and IL-33, in a mouse model of food allergy initially sensitized cutaneously with the food antigen. Mice were epicutaneously sensitized with the food antigen ovalbumin (OVA) followed by oral challenge with OVA. Epicutaneously sensitized mice produced OVA-specific IgE and developed IgE-dependent anaphylaxis after oral challenge. Basophil-depleted or TSLP-receptor-deficient mice did not produce OVA-specific IgE and were protected from oral challenge-induced anaphylaxis. IL-33-deficient mice produced normal levels of OVA-specific IgE. However, IL-33-deficient mice and mice treated with recombinant soluble IL-33 receptor were protected from anaphylaxis. Thus, basophils and TSLP have pivotal roles in Th2 development in the skin during the sensitization phase of food allergy. In contrast, while IL-33 is dispensable for promoting cutaneous antigen sensitization, the cytokine is essential for inducing IgE-dependent anaphylaxis in the gut.

Topics: Allergens; Anaphylaxis; Animals; Basophils; Cytokines; Disease Models, Animal; Female; Food Hypersensitivity; Immunoglobulin E; Interleukin-33; Interleukins; Mice; Mice, Knockout; Ovalbumin; Skin; Th2 Cells; Thymic Stromal Lymphopoietin

Considering the increasing numbers of patients suffering from food allergy (FA) as well as the great variety of novel foods and food compositions, an unmet need exists for the development of preclinical approaches to characterize the allergenic potential of proteins. The aim of our study was to evaluate the allergenicity of different food allergens in a rat model.. Brown Norway rats were sensitized to protein extracts (RuBisCO, apple, soy, peanut, garden pea) or ovalbumin (OVA) combined with Bordetella pertussis and aluminium hydroxide, followed by oral allergen challenges.. Allergen-specific serum immunoglobulin production and the proliferation of mononuclear cells from spleen confirmed sensitization. To assess functional alterations in the gut, intestinal permeability was measured, which increased in sensitized and challenged animals compared to non-sensitized controls. Allergens with high allergenic potential (peanut, OVA, soy) caused a stronger immunological response than allergens with low allergenic potential, such as RuBisCO and apple. Moreover, the immunological responses were reduced when using boiled instead of raw soy and pea proteins.. This model mimics key features of FA and facilitates investigating the allergenicity of allergens in novel food or food compositions in vivo.

Topics: Administration, Oral; Allergens; Animals; Bordetella pertussis; Cell Proliferation; Cells, Cultured; Disease Models, Animal; Food; Food Hypersensitivity; Immunoglobulins; Leukocytes, Mononuclear; Male; Ovalbumin; Permeability; Rats; Rats, Inbred BN; Spleen

Polyphenols, the potent plant secondary metabolites, have beneficial effects on human health, but the mechanism(s) by which these effects are exerted is not well understood. Here, we present the detailed analysis of the interactions between the major green tea catechin, epigallo-catechin 3-gallate (EGCG), and the major dietary protein and allergen, ovalbumin (OVA). We show that EGCG binds to the pocket that partly overlaps with the previously identified IgE-binding region in OVA, and that this interaction induces structural changes in the allergen. Moreover, our ex vivo studies reveal that OVA binds IgE and stimulates degranulation of basophils, and that its uptake by monocytes proceeds at a slower rate in the presence of EGCG. This study provides further evidence in support of the proposed mechanism by which EGCG interactions with the food allergens contribute to its diverse biological activities and may impair antigen uptake by antigen-presenting cells.

Topics: Allergens; Antigen-Presenting Cells; Basophils; Catechin; Circular Dichroism; Egg White; Electrophoresis, Polyacrylamide Gel; Food Hypersensitivity; Humans; Immunoglobulin E; Monocytes; Ovalbumin; Polyphenols; Protein Binding; Protein Conformation; Spectrometry, Fluorescence; Tea

The food contaminant bisphenol A (BPA) is pointed out as a risk factor in development of food allergy and food intolerance, two adverse food reactions increasing worldwide. We evaluated the consequences of perinatal exposure to low doses of BPA on immune-specific response to the food antigen ovalbumin (OVA) at adulthood. Perinatal exposure to BPA (0.5, 5, or 50 μg/kg/d) from 15th day of gravidity to pups weaning resulted in an increase of anti-OVA IgG titers at all BPA dosages in OVA-tolerized rats, and at 5 μg/kg/d in OVA-immunized rats compared to control rats treated with vehicle. In BPA-treated and OVA-tolerized rats, increased anti-OVA IgG titers were associated with higher IFNγ secretion by the spleen. This result is in accordance with the increase of activated CD4(+)CD44(high)CD62L(low) T lymphocytes observed in spleen of BPA-exposed rats compared to controls. Finally, when BPA-treated OVA-tolerized rats were orally challenged with OVA, colonic inflammation occurred, with neutrophil infiltration, increased IFNγ, and decreased TGFβ. We show that perinatal exposure to BPA altered oral tolerance and immunization to dietary antigens (OVA). In summary, the naive immune system of neonate is vulnerable to low doses of BPA that trigger food intolerance later in life.

Topics: Aging; Animals; Benzhydryl Compounds; Endocrine Disruptors; Female; Food Hypersensitivity; Immune System; Ovalbumin; Phenols; Pregnancy; Pregnancy, Animal; Prenatal Exposure Delayed Effects; Rats, Wistar; T-Lymphocytes; Weaning

The prevalence of food allergy is rising in the western world. Allergen restriction is the chosen treatment in this condition, but continuous ingestion of the antigen has shown positive results in clinical trials. In a previous study, we have shown several allergic and metabolic alterations after 7 days of ovalbumin (OVA) ingestion by sensitized mice. The aim of this study was to investigate whether prolonged ingestion of antigen by sensitized mice would reverse the metabolic consequences caused by experimental food allergy. For this, allergic and metabolic parameters were analysed after prolonged ingestion of an OVA diet by OVA-sensitized mice. As shown previously, after 7 days of OVA consumption, sensitized mice showed increased serum levels of anti-OVA immunoglobulin (Ig)E and IgG1, aversion to the antigen ingestion, marked body and adipose tissue weight loss, followed by adipose tissue inflammation and decreased serum levels of adipokines, glucose and triglycerides. However, after 14 days of oral challenge, sensitized mice showed an anti-OVA IgE level similar to the mice that were only sensitized, but the specific IgG1 did not change. With this prolonged ingestion of OVA, sensitized mice were protected from OVA-induced anaphylaxis when the antigen was given systemically at a dose of 2 mg/animal. Moreover, various parameters analysed were significantly ameliorated, including adipose tissue inflammation, body and adipose tissue loss, as well as serum levels of adipokines and triglycerides. Therefore, our data suggest that prolonged ingestion of OVA by sensitized mice results in an improvement of the metabolic consequences caused by experimental food allergy.

Topics: Adipose Tissue; Anaphylaxis; Animal Feed; Animals; Food Hypersensitivity; Glucose; Immunization; Immunoglobulin E; Male; Mice; Mice, Inbred BALB C; Ovalbumin; Weight Loss

Neuro-immune interaction in the gut is substantially involved in the maintenance of intestinal immune homeostasis and the pathology of intestinal immune diseases. We have previously demonstrated that mucosal mast cells and nerve fibers containing CGRP, a neurotransmitter of intrinsic enteric sensory neurons, are markedly increased and exist in close proximity to each other in the colon of food allergy (FA) mice. In the present study, a CGRP-receptor antagonist BIBN4096BS significantly alleviated allergic symptoms in the murine FA model. In addition, the elevated numbers of mucosal mast cells in the proximal colon of FA mice were significantly decreased in that of BIBN4096BS-treated FA mice. Thus, we investigated the effects of CGRP on calcium-independent process in degranulation of mucosal mast cells since CGRP increases intracellular cAMP levels, but not Ca(2+) concentration. CGRP did not alter a calcium ionophore A23187-increased cytosolic Ca(2+) concentration in mucosal-type bone marrow-derived mast cells (mBMMCs), but did augment microtubule reorganization in resting and A23187-activated mBMMCs. Furthermore, CGRP alone failed to cause the degranulation of mBMMCs, but CGRP significantly enhanced the degranulation of mBMMCs induced by A23187. Together, these data indicate that CGRP- enhanced microtubule reorganization augments IgE-independent/non-antigenic stimuli-induced mucosal mast cell degranulation, thereby contributing to the development of FA.

Topics: Animals; Calcitonin Gene-Related Peptide; Calcitonin Gene-Related Peptide Receptor Antagonists; Diarrhea; Disease Models, Animal; Food Hypersensitivity; Male; Mast Cells; Mastocytosis; Mice; Mice, Inbred BALB C; Microtubules; Neurotransmitter Agents; Ovalbumin; Piperazines; Quinazolines; Sensory Receptor Cells

Proteinogenic wine fining agents are hidden allergens and could present a risk for consumers with allergies. Therefore, the European Parliament adopted Directive 2003/89/EC amending Directive 2000/13/EC to declare ingredients, contaminations and processing aids that are known to trigger allergic reactions. The Amendment Regulation (EU) 1266/2010 excluded the labelling of wines which are processed with hen's egg and products thereof until 30 June 2012 to get more scientific findings. After 1 July 2012 wine fining agents have to be declared if above 0.25 mg l(-1) (Regulation (EU) 579/2012 in conjunction with article 120 g of Regulation (EU) 1234/2007). The Organisation International de la Vigne et du Vin (OIV) advises this limit of detection (LOD) for potential allergenic residues of proteins. Wine fining agents are processing aids and according to the wine producer's knowledge will be removed after coagulation by filtration or other production steps. Due to lack of scientific data, residues of fining agents in the final product could not be excluded. In this risk assessment, highly sensitive ELISA methods for ovalbumin of known origin for wine have been developed. The objective was to investigate the presence of allergen residues in wine after certain technological treatments were applied to remove the wine fining agents. For all developed ELISA methods the LODs are in the low µg l(-1) range between 5 and 10 µg l(-1) fining agent, whereas the LOQ varies between 5 and 80 µg l(-1) fining agent. The results of the investigation of well-known wines and fining agents demonstrate that white wines fined with white or ovalbumin from hen's egg could retain allergens. The use of certain technological procedures during wine processing leads to different results. In white wine, bentonite or sheet filtration followed by sterile filtration lead to wines containing no detectable amounts of ovalbumin. In red wine, especially the final sterile filtration removes the fining agents.

Topics: Adult; Allergens; Enzyme-Linked Immunosorbent Assay; Female; Food Contamination; Food Hypersensitivity; Humans; Limit of Detection; Middle Aged; Ovalbumin; Wine

To improve the efficacy and safety of tolerance induction for food allergies, identifying the tissues responsible for inducing intestinal inflammation and subsequent oral tolerance is important. We used OVA23-3 mice, which express an ovalbumin-specific T-cell receptor, to elucidate the roles of local and systemic immune tissues in intestinal inflammation.. OVA23-3 mice developed marked enteropathy after consuming a diet containing egg white (EW diet) for 10 days but overcame the enteropathy (despite continued moderate inflammation) after receiving EW diet for a total of 28 days. Injecting mice with anti-IL-4 antibody or cyclosporine A confirmed the involvement of Th2 cells in the development of the enteropathy. To assess the individual contributions of Peyer's patches (PPs), mesenteric lymph nodes (MLNs), and the spleen to the generation of effector CD4+ T-cells, we analyzed the IL-4 production, proliferation in response to ovalbumin, and CD4+ T-cell numbers of these tissues. EW feeding for 10 days induced significant IL-4 production in PPs, the infiltration of numerous CD4+ T-cells into MLNs, and a decrease in CD4+ T-cell numbers in spleen. On day 28, CD4+ T-cells from all tissues had attenuated responses to ovalbumin, suggesting tolerance acquisition, although MLN CD4+ T-cells still maintained IL-4 production with proliferation. In addition, removal of MLNs but not the spleen decreased the severity of enteropathy and PP-disrupted mice showed delayed onset of EW-induced inflammatory responses. Disruption of peripheral lymphoid tissues or of both PPs and MLNs almost completely prevented the enteropathy.. PPs and MLNs coordinately promote enteropathy by generating effector T-cells during the initial and exacerbated phases, respectively; the spleen is dispensable for enteropathy and shows tolerogenic responses throughout EW-feeding. The regulation of PPs may suppress the initiation of intestinal inflammation, subsequently restricting MLNs and inhibiting the progression of food-allergic enteropathy.

Topics: Animal Feed; Animals; CD4-Positive T-Lymphocytes; Disease Models, Animal; Egg White; Female; Food Hypersensitivity; Interleukin-4; Intestinal Diseases; Lymph Nodes; Male; Mesentery; Mice; Ovalbumin; Peyer's Patches; Spleen

Atopic march refers to the typical transition from a food allergy in early childhood to allergic asthma in older children and adults. However the precise interplay of events involving gut, skin and pulmonary inflammation in this process is not completely understood.. To develop a mouse model of mixed food and respiratory allergy mimicking the atopic march and better understand the impact of food allergies on asthma.. Food allergy to ovalbumin (OVA) was induced through intra-peritoneal sensitization and intra-gastric challenge, and/or a respiratory allergy to house dust mite (HDM) was obtained through percutaneous sensitization and intra-nasal challenges with dermatophagoides farinae (Der f) extract. Digestive, respiratory and systemic parameters were analyzed.. OVA-mediated gut allergy was associated with an increase in jejunum permeability, and a worsening of Der f-induced asthma with stronger airway hyperresponsiveness and pulmonary cell infiltration, notably eosinophils. There was overproduction of the pro-eosinophil chemokine RANTES in broncho-alveolar lavages associated with an enhanced Th2 cytokine secretion and increased total and Der f-specific IgE when the two allergies were present. Both AHR and lung inflammation increased after a second pulmonary challenge.. Gut sensitization to OVA amplifies Der f-induced asthma in mice.

Topics: Animals; Antigens, Dermatophagoides; Arthropod Proteins; Asthma; Bronchial Hyperreactivity; Bronchoconstriction; Chemokine CCL5; Disease Models, Animal; Female; Food Hypersensitivity; Immunoglobulin E; Intestinal Mucosa; Intestines; Lung; Mice, Inbred BALB C; Ovalbumin; Permeability; Pneumonia; Pulmonary Eosinophilia; Th2 Cells; Time Factors

The incidence of food allergy has increased in recent years and there is no effective way to treat it except strict dietary avoidance and rapid medical treatment in case of accidental exposure. Oral tolerance, as a new method, has shown great promise as an alternative approach to prevention and treatment for allergic disease. It was reported that all-trans retinoic acid (atRA) plays an important role in inducing oral tolerance in vitro. Our study aimed to investigate the immunological effect of different doses of atRA on ovalbumin (OVA) allergic BALB/c mice.. BALB/c mice were sensitized by intraperitoneal injection with OVA to establish allergic animal model. According to the dose of atRA given, 40 OVA allergic BALB/c mice were divided into 4 groups: the mice in high dose group were treated with 100 mg/kg atRA (atRA-H), those in median dose group were treated with 50 mg/kg atRA (atRA-M), those in low dose group were treated with 20 mg/kg atRA (atRA-L) and the mice in control group were given vehicle-soy oil only (CTR). After 12 days of atRA intervention, weight was measured, the mice were checked for diarrhea , and intestinal histology was observed after hematoxylin and eosin staining. The level of OVA-IgE in serum, total IgA and OVA-IgA in feces were measured by ELISA. The percentage of CD4⁺ CD25⁺ FoxP3⁺ T cells in CD4⁺ T cells in mesenteric lymph node was detected by flow cytometry.. Compared with that of CTR group, the level of OVA-IgE in serum (1.221 ± 0.367 vs. 0.793 ± 0.616) and OVA-IgA (1.573 ± 0.656 vs. 0.905 ± 0.279) in feces decreased significantly (P = 0.006 and 0.012, respectively) without weight and intestinal histology changes after low dose of atRA administration. However, there was no significant difference in the percentage of CD4⁺ CD25⁺ FoxP3⁺ T cells in CD4⁺ T cells in mesenteric lymph node (10.641 ± 1.218 vs. 10.936 ± 0.954) between atRA-L and CTR group (P > 0.05). While in animals with high and median dose of atRA administration, no immunologic improvement was found, instead, there was weight loss and intestinal mucosal damage.. Low dose of atRA intervention seems to induce immune suppression in vivo resulting in positive effects on OVA allergic mice. However, median and high dose atRA had no therapeutic effect on OVA allergic mice.

Topics: Animals; CD4-Positive T-Lymphocytes; Food Hypersensitivity; Immune Tolerance; Lymph Nodes; Mice; Mice, Inbred BALB C; Ovalbumin; T-Lymphocytes, Regulatory; Tretinoin

The intestinal tract plays an important role in food allergy and the intestinal mucosa barrier is critical for maintenance of its function. The underlying mechanisms of how food allergens modulate the intestinal permeability in inducing intestinal food allergy remain elusive.. The aim of this study was to explore the mechanism of how food allergens influence the function of intestinal barrier and induce intestinal food allergy.. Ovalbumin (OVA) was chosen to establish intestinal food allergy models in juvenile and adult rats that were confirmed by IgE and IgG assay. Intestinal tissue morphology was analyzed by HE staining. Intestinal permeability was dynamically monitored using a Lactulose (L)-Mannitol (M) assay. The morphology of the tight junctions in the intestinal mucosa barrier were analyzed under TEM. The expression of key molecules in tight junction regulation was evaluated by Real-time PCR.. We found: 1) The sensitization rate in juvenile rats was higher than in adult rats; 2) Intestine fluff erosion was more serious in juvenile rats than in adult rats in the duodenum and ileum; 3) Intestinal permeability was severely damaged, according to the results of the Lactulose (L)-Mannitol (M) assay; 4) Tight junction damage on the mucosal barrier was observed; Real-time PCR results showed that the expression of some key molecules that are involved in tight junction regulation was also affected.. Our data suggested that the allergy sensitization rate of Ovalbumin (OVA) in the juvenile group is higher than in adults and food allergens may increase intestinal mucosal permeability through intestinal tight junction regulation in inducing intestinal food allergy.

Topics: Age Factors; Allergens; Animals; Disease Models, Animal; Food Hypersensitivity; Immunohistochemistry; Intestinal Mucosa; Intestines; Microscopy, Electron, Transmission; Ovalbumin; Permeability; Rats; Real-Time Polymerase Chain Reaction; Tight Junction Proteins; Tight Junctions

Allergy is a skewed T helper (Th)2 polarization response in the body; its treatment is not satisfactory currently. Oral tolerance dysfunction plays a critical role in the pathogenesis of allergy. The present study aims to restore the breached intestinal tolerance with an artificial adduct of a measles virus C protein-derived small peptide (MVCP) and a model antigen, ovalbumin (MOA), and to observe the effect of MOA on inhibition of intestinal allergy in a mouse model. The MOA was formed by the MVCP and ovalbumin. The effect of MOA on regulation of the properties of dendritic cells (DC) and CD4(+) T cells was observed with a cell culture model and a mouse model of the gut Th2 pattern inflammation. After treatment with MOA, mouse intestinal DCs showed high levels of aldehyde dehydrogenase (ALDH) activity and expressed transforming growth factor (TGF)-beta; the frequency of Treg in the intestine was also significantly increased. The treatment with MOA efficiently suppressed the antigen-specific Th2 pattern inflammation in the intestine. Administration with the MOA can induce the development of antigen-specific oral tolerance and inhibit the antigen-specific allergic reaction in the intestine. The adduct of MOA has the therapeutic potential for the allergen related immune inflammation.

Topics: Aldehyde Dehydrogenase; Allergens; Animals; Antigens; CD4-Positive T-Lymphocytes; Dendritic Cells; Food Hypersensitivity; Immune Tolerance; Inflammation; Intestines; Measles virus; Mice; Mice, Inbred BALB C; Ovalbumin; Peptides; T-Lymphocytes, Regulatory; Th2 Cells; Transforming Growth Factor beta; Viral Proteins

Scutellaria baicalensis Georgi (skullcap) has been widely used as a dietary ingredient. The purpose of this study was to reveal novel function of skullcap and its mechanism on allergen permeation in intestinal epithelial cells. Intestinal epithelial Caco-2 cell monolayers were used to evaluate the inhibitory effect of skullcap on ovalbumin (OVA) permeation by measuring transepithelial electrical resistance (TEER) and the quantity of permeated OVA. TEER increased and the OVA flux decreased in a dose-dependent manner through up-regulating tight junction-related proteins in cells incubated with increasing concentrations of skullcap extract. In the in vivo study, the amounts of OVA from orally ingested albumen reduced on administration of the skullcap extract. We also revealed for the first time that the active component of skullcap extract for inhibition of OVA permeation was baicalein. These findings demonstrated that skullcap extract might attenuate a food allergic response by inhibiting allergen permeation in vitro and in vivo.

Topics: Allergens; Animals; Biological Transport; Caco-2 Cells; Cell Membrane Permeability; Down-Regulation; Food Hypersensitivity; Humans; Male; Mice, Inbred ICR; Ovalbumin; Plant Extracts; Scutellaria baicalensis

The aim of this study was to investigate the effect of maternal consumption of organically or conventionally produced feed on immunological biomarkers and their offsprings' response to a novel dietary antigen. First-generation rats were fed plant-based diets from two different cultivation systems (organic or conventional) or a chow. Second-generation rats were exposed to ovalbumin (OVA) via their mother's milk and subsequently challenged with OVA after weaning onto the chow diet. In the chow diet group feeding the dams OVA resulted in suppression of the pups' anti-OVA antibody response to the OVA challenge (total OVA-specific IgG was 197 for the OVA-treated chow diet group and 823 for the control chow diet group (arbitrary ELISA units)). In contrast, OVA exposure of the dams from the plant-based dietary groups did not result in a similar suppression. Cultivation system had no effect on the immunological biomarkers, except for a higher spleen prostaglandin E2 (PGE2) concentration in pups originating from dams fed the conventional plant-based diet (223 ng/L) than from those fed the organic plant-based diet (189 ng/L).

Topics: Animal Feed; Animals; Antigens; Diet; Dinoprostone; Female; Food; Food Hypersensitivity; Food, Organic; Immune Tolerance; Immunization, Passive; Immunoglobulin G; Maternal-Fetal Exchange; Ovalbumin; Plants, Edible; Pregnancy; Rats; Rats, Wistar; Spleen; Tumor Necrosis Factor-alpha

The importance of Ca(2+) influx via store-operated calcium channels (SOCs) leading to mast cell degranulation is well known in allergic disease. However, the underlying mechanisms are not fully understood. With food-allergic rat model, the morphology of degranulated mast cell was analysed by toluidine blue stain and electron microscope. Ca(2+) influx via SOCs was checked by Ca(2+) imaging confocal microscope. Furthermore, the mRNA and protein expression of SOCs subunits were investigated using qPCR and Western blot. We found that ovalbumin (OVA) challenge significantly increased the levels of Th2 cytokines and OVA-specific IgE in allergic animals. Parallel to mast cell activation, the levels of histamine in serum and supernatant of rat peritoneal lavage solution were remarkably increased after OVA treatment. Moreover, the Ca(2+) entry through SOCs evoked by thapsigargin was increased in OVA-challenged group. The mRNA and protein expressions of SOC subunits, stromal interaction molecule 1 (STIM1) and Orail (calcium-release-activated calcium channel protein 1), were dramatically elevated under food-allergic condition. Administration of Ebselen, a scavenger of reactive oxygen species (ROS), significantly attenuated OVA sensitization-induced intracellular Ca(2+) rise and upregulation of SOCs subunit expressions. Intriguingly, pretreatment with PI3K-specific inhibitor (Wortmannin) partially abolished the production of ROS and subsequent elevation of SOCs activity and their subunit expressions. Taken together, these results imply that enhancement of SOC-mediated Ca(2+) influx induces mast cell activation, contributing to the pathogenesis of OVA-stimulated food allergy. PI3K-dependent ROS generation involves in modulating the activity of SOCs by increasing the expressions of their subunit.

Topics: Animals; Calcium; Calcium Channels; Cell Degranulation; Female; Food Hypersensitivity; Immunoglobulin E; Mast Cells; Membrane Glycoproteins; ORAI1 Protein; Ovalbumin; Phosphatidylinositol 3-Kinases; Rats; Rats, Inbred BN; Reactive Oxygen Species; Stromal Interaction Molecule 1; Th2 Cells; Up-Regulation

To explore the effect of food allergy (FA) on the development of visceralgia sensibility and the substance P (SP) expression in colon of developing rats with FA.. Three-week old female Sprague-Dawley (SD) rats were randomly divided into two groups (n = 10 in each). The rats in FA group were sensitized with ovalbumin (OVA) 40 µg and Al(OH)3 1 mg suspension solution (0.2 ml) intraperitoneal (i.p.) injection on day 0, only OVA 40 µg solution i.p. on day 2, 4, 7, 9, 11, respectively, and the rats were challenged by gavage with OVA solution 30 mg (2 ml) on day 20, 24, 28, 30. The rats in non-sensitized (NS) group were not challenged except handled in the same ways. The serum OVA-IgE were determined by enzyme-linked immuno sorbent assay (ELISA) on day 0, 30. Jejunum segments were used to observe morphological structure, the expression of eosinophils, and the density and the percentage of degranulation of mast cells (MC). The rats were appraised for the pain sensibility of intestinal tract under colorectal distension irritation by the electrophysiological method on external oblique in the 18-24 hr after the last challenge. Colons were used to analyze the expression of SP through immunohistochemical staining and computer image analyzing system.. The serum OVA-IgE concentration and the eosinophils, mast cell, the percentage of mast cells degranulation in FA group were more than NS group (P < 0.01). The amplitudes of spike external oblique muscle of abdomen (EOMA, µV) of the FA group under the colorectal distension (CRD) pressures at 0, 15, 30, 45, 60, 75 mm Hg were (17.74 ± 0.72), (18.63 ± 1.72), (22.55 ± 1.70), (28.63 ± 7.00), (33.97 ± 7.34), (37.26 ± 8.40), and (17.43 ± 1.18), (17.27 ± 1.16), (17.73 ± 1.42), (19.55 ± 3.54), (23.29 ± 5.46), (25.20 ± 4.75) in NS group. With the CRD pressure increased, the amplitudes of spike EOMA increased significantly. There were significant differences between groups under the CRD pressures at 30, 45, 60, 75 mm Hg (F = 47.470, 13.367, 13.317, 15.390, P < 0.01). The expressions of colons SP in FA group and NS group are 247.12 ± 90.83 and 103.90 ± 58.94, respectively (t = 4.183, P < 0.01).. Sensitization through i.p. pathway and challenge by gavage with OVA in early life could result in FA in young SD rats. FA in early life enabled the amplitudes of spike EOMA and the expression of colons SP increase significantly. It may be related to increase in amount and degranulation of MC and SP abnormal expression in colon, which could lead to the development of visceralgia sensibility.

Topics: Animals; Colonic Diseases, Functional; Disease Models, Animal; Electrophysiology; Female; Food Hypersensitivity; Hyperalgesia; Intestinal Mucosa; Mast Cells; Ovalbumin; Pain Threshold; Rats; Rats, Sprague-Dawley; Stress, Psychological; Substance P

Food allergy is a severe human disease with imminent risk of life. Cissampelos sympodialis (Menispermaceae) is a native Brazilian plant used in Brazilian folk medicine for the treatment of respiratory allergies. In this study the experimental model of food allergy induced by ovalbumin (OVA) was used to determine whether the alcoholic extract of the plant (AFL) and its alkaloids match a therapeutic approach for this disease. Animal weight, diarrhea, OVA-specific IgE levels, inflammatory cell and cytokine profiles, mucus production and proportion of T cells on the mesenteric lymph node (MLN) were evaluated. Warifteine (W) or methyl-warifteine (MW) alkaloids slightly improve diarrhea score independently of AFL and all treatments decreased the OVA-specific IgE levels. Stimulated mesenteric lymph node (MLN) cells in the presence of the alkaloids diminished the IL-12p70 levels independently of IFN-γ or IL-13 secretion. The alkaloids increased the number of Treg cells on MLN and reduced the number of eosinophils and mast cells as well as mucus production in the gut. Therefore, the alkaloids modulate the immune response in food allergy by increasing regulatory T cells in MLN independently of Th1 or Th2 profiles.

Topics: Alkaloids; Animals; Antibody Formation; Cells, Cultured; Cissampelos; Cytokines; Disease Models, Animal; Eosinophils; Female; Food Hypersensitivity; Humans; Immunoglobulin E; Mast Cells; Mice; Mice, Inbred BALB C; Ovalbumin; Phytotherapy; Plant Extracts; T-Lymphocyte Subsets; T-Lymphocytes, Regulatory

Arecae semen, the dried slice of areca nuts, is a traditional Chinese medicine used to treat intestinal parasitosis, rectal tenesmus and diarrhea. Areca nuts contain a rich amount of polyphenols that have been shown to modulate the functionality of mast cells and T cells. The objective of this study is to investigate the effect of polyphenol-enriched areca nut extracts (PANE) against food allergy, a T cell-mediated immune disorder.. BALB/c mice were left untreated or administered with PANE (0.05% and 0.1%) via drinking water throughout the entire experiment. The mice were sensitized with ovalbumin (OVA) twice by intraperitoneal injection, and then repeatedly challenged with OVA by gavage to induce food allergic responses.. PANE administration attenuated OVA-induced allergic responses, including the occurrence of diarrhea and the infiltration and degranulation of mast cells in the duodenum. The serum level of OVA-specific IgE and the expression of interleukin-4 in the duodenum were suppressed by PANE treatment. In addition, PANE administration induced Gr-1+, IL-10+ and Gr-1+IL-10+ cells in the duodenum.. These results demonstrate that oral intake of areca-derived polyphenols attenuates food allergic responses accompanied with a decreased Th2 immunity and an enhanced induction of functional myeloid-derived suppressor cells.

Topics: Animals; Areca; Drugs, Chinese Herbal; Food Hypersensitivity; Humans; Immunity, Cellular; Immunoglobulin E; Interleukin-10; Interleukin-4; Male; Mice; Mice, Inbred BALB C; Ovalbumin; Polyphenols; Th2 Cells

The prevalence of food allergy has increased dramatically during the last three decades, but currently there was no effective therapy except avoidance of allergen. This study aimed to investigate the effect of a modified Chinese herbal formula, Formula-3, on mast cell degranulation and the anti-allergic activity in both animal and cell models.. With OVA-sensitized food allergic model in Brown-Norway rats, we checked tissue injury in the small intestines by H&E staining. The Th2 cytokine levels and IgE production in serum or supernatant of the intestinal mucosa homogenates were analyzed by ELISA. Meanwhile, rat peritoneal mast cell activation and degranulation were examined by Toluidine Blue Stain and the release of histamine was measured. Furthermore, the regulation of Formula-3 on Ca(2+) mobilization was investigated by probing intracellular Ca(2+) with fluo-4 fluorescence. The direct effect of Formula-3 on mast cell stabilization was also studied in RBL-2H3 cell line.. In vivo Formula-3 administration significantly reduced tissue damage in the small intestines of rat and suppressed Th2 cytokine secretion and IgE production. We demonstrated that Formula-3 treatment significantly suppressed FcεR1-mediated mast cell degranulation no matter in OVA-challenged allergic rats or IgE-sensitized RBL-2H3 cell line. Furthermore, Formula-3 significantly decreased Ca(2+) influx through store-operated calcium channels (SOCs) evoked by dinitrophenyl-BSA or thapsigargin in mast cells.. Taken together, our data indicate that Formula-3 stabilizes mast cells by suppressing FcεR1-induced Ca(2+) mobilization mainly through inhibiting Ca(2+) entry via SOCs, thus exerting a protective effect against OVA-sensitized food allergy.

Topics: Allergens; Animals; Anti-Allergic Agents; Calcium; Cell Degranulation; Cytokines; Drugs, Chinese Herbal; Female; Food Hypersensitivity; Immunoglobulin E; Intestine, Small; Mast Cells; Ovalbumin; Rats; Rats, Inbred Strains

To investigate the effect of polydatin (PD), a resveratrol glucoside, on mast cell degranulation and anti-allergic activity.. After the rats were orally sensitized with ovalbumin (OVA) for 48 d and underwent PD treatment for 4 d, all the rats were stimulated by 100 mg/mL OVA for 24 h and then sacrificed for the following experiments. The small intestines from all the groups were prepared for morphology examination by hematoxylin and eosin staining. We also used a smooth muscle organ bath to evaluate the motility of the small intestines. The OVA-specific immunoglobulin E (IgE) production and interleukin-4 (IL-4) levels in serum or supernatant of intestinal mucosa homogenates were analyzed by enzyme-linked immunosorbent assay (ELISA). Using toluidine blue stain, the activation and degranulation of isolated rat peritoneal mast cells (RPMCs) were analyzed. Release of histamine from RPMCs was measured by ELISA, and regulation of PD on intracellular Ca(2+) mobilization was investigated by probing intracellular Ca(2+) with fluo-4 fluorescent dye, with the signal recorded and analyzed.. We found that intragastric treatment with PD significantly reduced loss of mucosal barrier integrity in the small intestine. However, OVA-sensitization caused significant hyperactivity in the small intestine of allergic rats, which was attenuated by PD administration by 42% (1.26 ± 0.13 g vs OVA 2.18 ± 0.21 g, P < 0.01). PD therapy also inhibited IgE production (3.95 ± 0.53 ng/mL vs OVA 4.53 ± 0.52 ng/mL, P < 0.05) by suppressing the secretion of Th2-type cytokine, IL-4, by 34% (38.58 ± 4.41 pg/mL vs OVA 58.15 ± 6.24 pg/mL, P < 0.01). The ratio of degranulated mast cells, as indicated by vehicles (at least five) around the cells, dramatically increased in the OVA group by 5.5 fold (63.50% ± 15.51% vs phosphate-buffered saline 11.15% ± 8.26%, P < 0.001) and fell by 65% after PD treatment (21.95% ± 4.37% vs OVA 63.50% ± 15.51%, P < 0.001). PD mediated attenuation of mast cell degranulation was further confirmed by decreased histamine levels in both serum (5.98 ± 0.17 vs OVA 6.67 ± 0.12, P < 0.05) and intestinal mucosa homogenates (5.83 ± 0.91 vs OVA 7.35 ± 0.97, P < 0.05). Furthermore, we demonstrated that administration with PD significantly decreased mast cell degranulation due to reduced Ca(2+) influx through store-operated calcium channels (SOCs) (2.35 ± 0.39 vs OVA 3.51 ± 0.38, P < 0.01).. Taken together, our data indicate that PD stabilizes mast cells by suppressing intracellular Ca(2+) mobilization, mainly through inhibiting Ca(2+) entry via SOCs, thus exerting a protective role against OVA-sensitized food allergy.

Topics: Animals; Calcium; Calcium Channels; Disease Models, Animal; Drugs, Chinese Herbal; Female; Food Hypersensitivity; Glucosides; Histamine; Immunoglobulin E; Interleukin-4; Intestine, Small; Mast Cells; Ovalbumin; Rats; Rats, Inbred BN; Stilbenes

Modified vaccinia virus Ankara (MVA)-encoding antigens are considered as safe vaccine candidates for various infectious diseases in humans. Here, we investigated the immune-modulating properties of MVA-encoding ovalbumin (MVA-OVA) on the allergen-specific immune response.. The immune-modulating properties of MVA-OVA were investigated using GM-CSF-differentiated BMDCs from C57BL/6 mice. OVA expression upon MVA-OVA infection of BMDCs was monitored. Activation and maturation markers on viable MVA-OVA-infected mDCs were analyzed by flow cytometry. Secretion of INF-γ, IL-2, and IL-10 was determined in a co-culture of BMDCs infected with wtMVA or MVA-OVA and OVA-specific OT-I CD8(+) and OT-II CD4(+ ) T cells. BALB/c mice were vaccinated with wtMVA, MVA-OVA, or PBS, sensitized to OVA/alum and challenged with a diet containing chicken egg white. OVA-specific IgE, IgG1, and IgG2a and cytokine secretion from mesenteric lymph node (MLN) cells were analyzed. Body weight, body temperature, food uptake, intestinal inflammation, and health condition of mice were monitored.. Infection with wtMVA and MVA-OVA induced comparable activation of mDCs. MVA-OVA-infected BMDCs expressed OVA and induced enhanced IFN-γ and IL-2 secretion from OVA-specific CD8(+ ) T cells in comparison with OVA, wtMVA, or OVA plus wtMVA. Prophylactic vaccination with MVA-OVA significantly repressed OVA-specific IgE, whereas OVA-specific IgG2a was induced. MVA-OVA vaccination suppressed TH 2 cytokine production in MLN cells and prevented the onset of allergic symptoms and inflammation in a mouse model of OVA-induced intestinal allergy.. Modified vaccinia virus Ankara-ovalbumin (MVA-OVA) vaccination induces a strong OVA-specific TH 1- immune response, likely mediated by the induction of IFN-γ and IgG2a. Finally, MVA-based vaccines need to be evaluated for their therapeutic potential in established allergy models.

Topics: Allergens; Animals; Bone Marrow Transplantation; Cells, Cultured; Coculture Techniques; Dendritic Cells; Disease Models, Animal; Food Hypersensitivity; Immunotherapy, Adoptive; Inflammation; Intestinal Mucosa; Mice; Mice, Inbred BALB C; Mice, Inbred C57BL; Ovalbumin; Th1 Cells; Vaccines, Synthetic; Vaccinia; Vaccinia virus; Viral Vaccines

We had found that orally administered Lactobacillus species were effective immune modulators in ovalbumin (OVA)-sensitized mice. To validate these findings, we investigated the effects of orally administered Lactobacillus brevis HY7401 in OVA-T cell receptor transgenic mice. This strain showed a tendency to induce Th1 cytokines and inhibit Th2 cytokines. All assayed isotypes of OVA-specific antibody were effectively reduced. Systemic anaphylaxis was also relatively reduced with the probiotic administration. These results reveal that L. brevis HY7401 might be useful to promote anti-allergic processes through oral administration.

Topics: Administration, Oral; Allergens; Animals; Cytokines; Diet; Disease Models, Animal; Food Hypersensitivity; Levilactobacillus brevis; Mice, Transgenic; Ovalbumin; Probiotics; Th1 Cells; Th2 Cells

Specific immunotherapy (SIT) is the only specific remedy for the treatment of allergic diseases currently. B cells are important immune cells in the immunity. The role of B cells in immune regulatory activities has not been fully understood yet. This study aims to elucidate the role of the thrombospondin (TSP)1-producing B cells in the immune regulatory role of SIT. The results showed that after SIT, the frequency of CD35(+) B cells was increased in the intestine of mice with food allergy. The CD35(+) B cells expressed TSP1 after exposure to specific antigens. Co-culture with the TSP1-producing CD35(+) B cells decreased the levels of CD80/CD86 in dendritic cells; the cells convert naïve CD4(+) T cells to regulatory T cells to inhibit allergic inflammation in the intestine.

Topics: Animals; B-Lymphocytes; B7-1 Antigen; B7-2 Antigen; Cholera Toxin; Dendritic Cells; Food Hypersensitivity; Immune Tolerance; Immunotherapy; Inflammation; Interleukin-13; Interleukin-4; Interleukin-5; Intestines; Lymphocyte Activation; Mice; Mice, Inbred BALB C; Ovalbumin; Receptors, Complement 3b; T-Lymphocytes, Regulatory; Thrombospondin 1

Haptens can bind to proteins to elicit antigenicity. Whether haptens play a role in the pathogenesis of food allergy remains to be investigated. This article aims to elucidate the role a hapten plays in food antigen-related T helper 2 (Th2) pattern inflammation in the intestine.. The effect of trinitrobenzene sulfonic acid (TNBS; as a hapten) on the properties of dendritic cells was assessed by a cell culture model. BALB/c mice were sensitized with a mixture of TNBS and ovalbumin (OVA; as a model antigen). Intestinal Th2 response, OVA-specific immunoglobulin E and histamine were analyzed with the mouse model. In addition to the infiltration of the intestinal inflammatory cells, cytokine expression profiles were determined.. TNBS increased the expression of T-cell immunoglobulin and mucin domain-4 and CD80 and decreased the levels of interleukin-12 in dendritic cells. Higher serum levels of OVA-specific immunoglobulin E, histamine expression and skewed antigen-specific Th2 polarization in the intestinal tissue were detected in mice sensitized with TNBS + OVA as compared with those treated with either OVA or TNBS alone. In addition, the TNBS-OVA-treated mice also showed an increased number of inflammatory cells, high levels of interleukin-4 and a decreased expression of interferon-γ in the lamina propria mononuclear cells.. Hapten TNBS can facilitate the initiation of food antigen-related Th2 pattern inflammation, such as food allergy, in the intestine.

Topics: Allergens; Animals; Cells, Cultured; Food Hypersensitivity; Haptens; Intestinal Mucosa; Male; Mice; Mice, Inbred BALB C; Ovalbumin; Th2 Cells; Trinitrobenzenesulfonic Acid

Sensitization to food antigen can occur through cutaneous exposure.. We sought to test the hypothesis that epicutaneous sensitization with food antigen predisposes to IgE-mediated anaphylaxis on oral allergen challenge.. BALB/c mice were epicutaneously sensitized by repeated application of ovalbumin (OVA) to tape-stripped skin over 7 weeks or orally immunized with OVA and cholera toxin (CT) weekly for 8 weeks and then orally challenged with OVA. Body temperature was monitored, and serum mouse mast cell protease 1 levels were determined after challenge. Tissue mast cell (MC) counts were examined by using chloroacetate esterase staining. Levels of serum OVA-specific IgE and IgG(1) antibodies and cytokines in supernatants of OVA-stimulated splenocytes were measured by means of ELISA. Serum IL-4 levels were measured by using an in vivo cytokine capture assay.. Epicutaneously sensitized mice exhibited expansion of connective tissue MCs in the jejunum, increased serum IL-4 levels, and systemic anaphylaxis after oral challenge, as evidenced by decreased body temperature and increased serum mouse mast cell protease 1 levels. Intestinal MC expansion and anaphylaxis were IgE dependent because they did not occur in epicutaneously sensitized IgE(-/-) mice. Mice orally immunized with OVA plus CT did not have increased serum IL-4 levels, expanded intestinal MCs, or anaphylaxis after oral challenge, despite OVA-specific IgE levels and splenocyte cytokine production in response to OVA stimulation, which were comparable with those of epicutaneously sensitized mice.. Epicutaneously sensitized mice, but not mice orally immunized with antigen plus CT, have expansion of intestinal MCs and IgE-mediated anaphylaxis after single oral antigen challenge. IgE is necessary but not sufficient for food anaphylaxis, and MC expansion in the gut can play an important role in the development of anaphylaxis.

Topics: Administration, Cutaneous; Allergens; Anaphylaxis; Animals; Antibodies; Antigens; Body Temperature; Chemokine CCL2; Cholera Toxin; Food Hypersensitivity; Immunoglobulin E; Immunoglobulin G; Interleukin-4; Jejunum; Mast Cells; Mice; Mice, Inbred BALB C; Ovalbumin; Skin

Shellfish hypersensitivity is among the most common food allergies. The allergens tropomyosin (TM) and arginine kinase (AK) from mud crab (Scylla paramamosain) were purified to homogeneity. BALB/c female mice were sensitized with TM and AK by intragastric administration. Mice treated with normal saline served as the negative control (NC) group.. Compared with NC group, mice that were treated with TM and AK developed reduced activity; meanwhile, their scratching behavior and specific-IgE level were increased. Specific-CD4 + T cells were significantly elevated in the splenocyte cultures of the mice upon TM and AK stimulation. However, compared with the positive control group (ovalbumin, OVA), there was no significant difference. The expression of IL-4 in culture cells stimulated by TM, AK, and OVA group showed significant differences from the NC group, respectively.. These results indicated that a BALB/c mouse model for sensitization to TM and AK from mud crab was successfully established, and the Th2 response was observed, displaying increased immunoglobulin E levels, together with the production of interleukin 4 and allergic symptoms.

Topics: Allergens; Animals; Arginine Kinase; Brachyura; CD4-Positive T-Lymphocytes; Cells, Cultured; Cytokines; Female; Food Hypersensitivity; Gene Expression Regulation; Immunoglobulin E; Mice; Mice, Inbred BALB C; Ovalbumin; Random Allocation; RNA, Messenger; Spleen; Tropomyosin

Utilizing a food allergy murine model, we have investigated the intrinsic antiallergic potential of the Lactococcus lactis NCC 2287 strain.. BALB/c mice were sensitized at weekly intervals with ovalbumin (OVA) plus cholera toxin (CT) by the oral route for 7 weeks. In this model, an oral challenge with a high dose of OVA at the end of the sensitization period leads to clinical symptoms. Lactococcus lactis NCC 2287 was given to mice via the drinking water during sensitization (prevention phase) or after sensitization (management phase).. Lactococcus lactis NCC 2287 administration to sensitized mice strikingly reduced allergic manifestations in the management phase upon challenge, when compared to control mice. No preventive effect was observed with the strain. Lactococcus lactis NCC 2287 significantly decreased relative expression levels of the Th-2 cytokine, IL-13, and associated chemokines CCL11 (eotaxin-1) and CCL17 (TARC) in the ileum. No effect was observed in the jejunum.. These results taken together designate Lactococcus lactis NCC 2287 as a candidate probiotic strain appropriate in the management of allergic symptoms.

Topics: Administration, Oral; Animals; Chemokine CCL11; Chemokine CCL17; Cholera Toxin; Enzyme-Linked Immunosorbent Assay; Female; Food Hypersensitivity; Ileum; Immunoglobulin E; Immunoglobulin G; Interleukin-13; Lactococcus lactis; Mice; Mice, Inbred BALB C; Ovalbumin; Probiotics

Sphingosine-1-phosphate (S1P) influences activation, migration and death of immune cells. Further, S1P was proposed to play a major role in the induction and promotion of allergic diseases. However, to date only limited information is available on the role of S1P in food allergy.. We aimed to investigate the role of sphingosine-kinase (SphK) 1 and 2, the enzymes responsible for endogenous S1P production, on the induction of food allergy.. Human epithelial colorectal CaCo2 cells stimulated in vitro with S1P revealed a decrease of transepithelial resistance and enhanced transport of FITC labeled OVA. We studied the effect of genetic deletion of the enzymes involved in S1P production on food allergy induction using a mouse model of food allergy based on intragastrically (i.g.) administered ovalbumin (OVA) with concomitant acid-suppression. Wild-type (WT), SphK1(-/-) and SphK2(-/-) mice immunized with OVA alone i.g. or intraperitoneally (i.p.) were used as negative or positive controls, respectively. SphK1- and SphK2-deficient mice fed with OVA under acid-suppression showed reduced induction of OVA specific IgE and IgG compared to WT mice, but had normal responses when immunized by the intraperitoneal route. Flow cytometric analysis of spleen cells revealed a significantly reduced proportion of CD4(+) effector T-cells in both SphK deficient animals after oral sensitization. This was accompanied by a reduced accumulation of mast cells in the gastric mucosa in SphK-deficient animals compared to WT mice. Furthermore, mouse mast cell protease-1 (mMCP-1) levels, an IgE-mediated anaphylaxis marker, were reliably elevated in allergic WT animals.. Modulation of the S1P homeostasis by deletion of either SphK1 or SphK2 alters the sensitization and effector phase of food allergy.

Topics: Administration, Oral; Animals; Caco-2 Cells; CD4-Positive T-Lymphocytes; Cell Movement; Chemokine CCL2; Disease Models, Animal; Food Hypersensitivity; Gastric Mucosa; Humans; Immunoglobulin E; Mast Cells; Mice; Mice, Inbred Strains; Mice, Knockout; Ovalbumin; Protein Tyrosine Phosphatase, Non-Receptor Type 11; Protein Tyrosine Phosphatase, Non-Receptor Type 6; Up-Regulation

The number of food allergy patients is increasing. Some children outgrow their food allergies through tolerance, whereas others remain susceptible throughout their lives. We aimed to contribute to food allergy therapeutics by understanding induction of oral tolerance in a murine food allergy model.. We modified an existing murine food allergy model by using ovalbumin (OVA) to induce oral tolerance, either by pretreating mice with OVA or by transferring mesenteric lymph node (MLN) cells or T cells derived from mice treated with OVA.. Pretreatment with OVA prevented food allergy, with complete suppression of OVA-specific immunoglobulin (Ig)E and IgA antibody production and interleukin (IL)-4, IL-10, and IL-9 mRNA expression. The proportion of regulatory T cells (Tregs) in MLN cells and expression of transforming growth factor-β mRNA increased. In the transfer model, anaphylaxis secondary to OVA intake was suppressed by transfer of whole MLN cells and Tregs from OVA-treated mice. However, OVA-specific IgE and IgA expressions were partially attenuated by transfer of antigen-specific and nonspecific Tregs, but not by whole MLN cells from OVA-treated mice. In the Treg transfer model, IL-4 and IL-10 mRNA expression decreased, but IL-9 mRNA expression increased.. We concluded that oral tolerance for food antigens is induced in two ways: (i) by initial exposure to antigen, or inherent tolerance, and (ii) by transfer of Tregs, or acquired tolerance. Because food allergies occur when inherent tolerance is absent, understanding of acquired tolerance is important for the development of therapies for food allergy.

Topics: Administration, Oral; Adoptive Transfer; Allergens; Anaphylaxis; Animals; Cytokines; Disease Models, Animal; Female; Food Hypersensitivity; Immune Tolerance; Immunoglobulin A; Immunoglobulin E; Mice; Mice, Inbred BALB C; Ovalbumin; T-Lymphocytes, Regulatory; Time Factors

We investigated the effect of dietary supplementation with n-3 PUFA (fish oil source) in an experimental model of food allergy. Mice were sensitized (allergic group) or not (nonallergic group) with OVA and were fed with OVA diet to induce allergy signals. Mice were fed with regular diet in which 7% of lipid content was provided by soybean (5% of n-3 PUFA) or fish (25% of n-3 PUFA) oil. Allergic group mice had increased serum levels of antiovalbumin IgE and IgG1 and changes in small intestine, characterized by an increased edema, number of rolling leukocytes in microcirculation, eosinophil infiltration, mucus production, and Paneth cell degranulation, in comparison to non-allergic group. All these inflammatory parameters were reduced in mice fed high-n-3-PUFA diet. Our data together suggest that diet supplementation with n-3 PUFA from fish oil may consist of a valid adjuvant in food allergy treatment.

Topics: Animals; Dietary Supplements; Fatty Acids, Omega-3; Female; Fish Oils; Food Hypersensitivity; Intestinal Mucosa; Mice; Mice, Inbred BALB C; Ovalbumin

There are conflicting data to support the practice of delaying the introduction of allergenic foods into the infant diet to prevent allergy development. This study investigated immune response development after early oral egg antigen (Ovalbumin; OVA) exposure in a rat pup model. Brown Norway (BN) rat pups were randomly allocated into groups: dam reared (DR), DR pups challenged daily (days 4-13) with oral OVA (DR + OVAc), DR pups challenged intermittently (on day 4, 10, 12, and 13) with oral OVA (DR + OVAi), formula-fed pups (FF), and FF pups challenged daily with oral OVA (FF + OVA). Immune parameters assessed included OVA-specific serum IgE, IgG1, and IgA. Ileal and splenic messenger ribonucleic acid (mRNA) expression of transforming growth factor-beta (TGF-β1), mothers against decapentaplegic (Smad) 2/4/7, and forkhead box P3 (Foxp3) were determined. Ileum was stained for TGF-β1 and Smad4. Results. Feeding OVA daily to DR pups maintained systemic and local gut antibody and immunoregulatory marker mRNA responses. Systemic TGF-β1 was lower in DR + OVAi pups compared to DR and DR + OVAc pups. Feeding OVA to FF pups resulted in significantly greater OVA-specific IgE and IgG1, and lower IgA and TGF-β1 and Smad expression compared to DR pups. Conclusions. Early daily OVA exposure in the presence of maternal milk maintains immune markers associated with a regulated immune response, preventing early allergic sensitization.

Topics: Animals; Body Weight; Food Hypersensitivity; Ileum; Immunity, Mucosal; Immunoglobulin A; Immunoglobulin E; Immunoglobulin G; Intestinal Mucosa; Ovalbumin; Rats; Rats, Inbred BN; RNA, Messenger; Smad Proteins; Spleen; Time Factors; Transforming Growth Factor beta1

Food-triggered anaphylaxis can encompass a variety of symptoms that affect multiple organ systems and can be life threatening. The molecular distinction between non-life-threatening and life-threatening modes of such anaphylaxis has not yet been delineated. In this study, we sought to identify the specific immune functions that regulate the severity of oral antigen-induced anaphylaxis. We thus developed an experimental mouse model in which repeated oral challenge of ovalbumin-primed mice induced an FcεRI- and IgE-dependent oral antigen-triggered anaphylaxis that involved multiple organ systems. Strikingly, the severity of the systemic symptoms of anaphylaxis (eg, hypothermia) positively correlated with the levels of intestinal mast cells (r = -0.53; P < 0.009). In addition, transgenic mice with both increased intestinal and normal systemic levels of mast cells showed increased severity of both intestinal and extra-intestinal symptoms of IgE-mediated passive as well as oral antigen- and IgE-triggered anaphylaxis. In conclusion, these observations indicate that the density of intestinal mast cells controls the severity of oral antigen-induced anaphylaxis. Thus, an awareness of intestinal mast cell levels in patients with food allergies may aid in determining their susceptibility to life-threatening anaphylaxis and may eventually aid in the treatment of food-triggered anaphylaxis.

Topics: Administration, Oral; Anaphylaxis; Animals; Antigens; Capillary Permeability; Cell Count; Diffusion Chambers, Culture; Disease Models, Animal; Food Hypersensitivity; Immunoglobulin E; Jejunum; Mast Cells; Mice; Mice, Inbred BALB C; Mice, Transgenic; Ovalbumin; Receptors, IgE; Severity of Illness Index

Allergen-specific immunotherapy has been anticipated to be a disease-modifying therapy for food allergies. We previously reported that CD8(+) regulatory T cells may prevent antigen-sensitized mice from developing allergic diarrhea. Because oligomannose-coated liposomes (OML) have been shown to induce MHC class I-restricted CD8(+) T cell responses, we analyzed the adjuvant activities of OML for inducing regulatory CD8(+) T cells and mucosal tolerogenic responses in allergen-sensitized mice.. The BALB/c mice that were previously sensitized to ovalbumin (OVA) were intranasally immunized with OVA-encased in OML (OVA-OML) or OVA-encased in non-coated liposomes (OVA-NL). We assessed allergic diarrhea induced by oral OVA administration, OVA-specific immunoglobulin production, and cytokine production in the intestines and mesenteric lymph nodes (MLNs).. Intranasal immunization with OVA-OML, but not OVA-NL, suppressed the development of allergic diarrhea. This was associated with in vitro Ag-induced IL-10 production and the in vivo expansion of CD8(+) CD28(-) and CD4(+) CD25(+) Foxp3(+) T cell populations among mesenteric lymph node mononuclear cells, and was significantly ablated by anti-SIGNR1 or anti-CR3 mAbs. Up-regulation of serum OVA-specific IgE was suppressed, whereas OVA-specific IgG1, IgG2a, and soluble IgA production were enhanced by intranasal administration of OVA-OML. Adoptive transfer of CD8(+) CD28(-) T cells but not CD28(+) CD8(+) T cells from the MLNs of OVA-OML-treated mice ameliorated the development of diarrhea.. These results suggest that intranasal immunization with Ag-encased OML may be an effective immunotherapy for food allergies, as it induces a subset of regulatory CD8(+) T cells as well as CD4(+) CD25(+) Foxp3(+) T cell and modulates humoral immune responses in allergen-sensitized mice.

Topics: Animals; CD8-Positive T-Lymphocytes; Desensitization, Immunologic; Diarrhea; Disease Models, Animal; Food Hypersensitivity; Humans; Liposomes; Mice; Mice, Inbred BALB C; Oligosaccharides; Ovalbumin; T-Lymphocytes, Regulatory; Treatment Outcome

The clinical manifestations of food allergy include diarrhea and systemic anaphylaxis (shock), which can occur together or by themselves in different subjects. Although ingested food antigens need to be absorbed to induce shock, it is not known whether they need to be absorbed to induce diarrhea.. We sought to identify mechanisms that determine whether food allergy induces diarrhea versus shock and determine whether diarrhea requires absorption of ingested antigens.. These issues were studied in mice in active, passive, and hybrid immunization models. The active model was used to determine the allergic diarrhea susceptibility of J chain- and polymeric immunoglobulin receptor-deficient mice, which are unable to secrete IgA. The hybrid model was used to determine whether intravenously administered antigen-specific IgG antibody, which is not secreted into the gut, can protect against allergic diarrhea, as well as shock.. Shock, but not diarrhea, was induced in naive mice by using intravenous IgE anti-trinitrophenyl (TNP) antibody, followed by oral TNP-BSA, whereas both were induced in mice presensitized with intraperitoneal ovalbumin/alum plus oral ovalbumin. More TNP-BSA was required to induce shock than diarrhea in presensitized mice, and intravenous IgG anti-TNP antibody, which is not secreted into the gut, protected these mice against both diarrhea and shock. Consistent with this, chicken ovalbumin-immunized J chain- and polymeric immunoglobulin receptor-deficient mice, which have high serum IgA levels but little intestinal IgA, resisted diarrhea induction.. Intestinal immunity and oral antigen dose determine whether diarrhea, systemic anaphylaxis, or both are induced, and ingested antigen must be absorbed to induce either response.

Topics: Anaphylaxis; Animals; Clinical Protocols; Diarrhea; Disease Models, Animal; Fatty Acid-Binding Proteins; Food Hypersensitivity; Humans; Immunization; Immunoglobulin J-Chains; Immunoglobulins; Interleukin-9; Mice; Mice, Inbred BALB C; Mice, Knockout; Mice, Transgenic; Ovalbumin; Promoter Regions, Genetic; Receptors, IgG; Receptors, Polymeric Immunoglobulin; Serum Albumin, Bovine; Trinitrobenzenes

It is important to evaluate the ability of novel proteins in food crops and products to elicit potentially harmful immunologic responses, including allergic hypersensitivity. We developed a novel mouse model of food allergy involving an oral challenge of a protein antigen after feeding of the antigen in combination with modulating factors often ingested in daily life, namely, dietary oil emulsion and salicylate. In the model, BALB/c mice were sensitized orally for three weeks with ovalbumin (OVA) in linoleic acid/lecithin emulsion, followed immediately by intraperitoneal injection of sodium salicylate. At the end of the sensitization, the incidence of mice positive for serum OVA-specific IgG1 but not IgE had significantly increased in the combined-sensitization group. After the 3-week sensitization, a single or double oral challenge with OVA effectively and significantly caused severe anaphylaxis, as compared with the groups sensitized with OVA in the emulsion or the vehicle alone. Moderate increase of plasma histamine and intestinal abnormality in histology was found only in the combined-sensitization group. Anaphylaxis symptoms in the sensitized mice were induced more by oral challenge than by intravenous challenge, suggesting a critical role for the mucosal system. This is the first model for successful induction of oral anaphylaxis in mice sensitized by feeding of food protein without adjuvant. It will be useful to elucidate the mechanism of food allergy and to detect modulating factors of oral allergy at sensitization using this model, which simulates real life conditions.

Topics: Administration, Oral; Allergens; Anaphylaxis; Animals; Disease Models, Animal; Emulsions; Female; Food Hypersensitivity; Immunoglobulin G; Intestine, Small; Lecithins; Linoleic Acid; Mice; Mice, Inbred BALB C; Ovalbumin; Salicylic Acid

With the development of genetically modified crops, there has been a growing interest in available approaches to assess the potential allergenicity of novel gene products. We were not sure whether Cry1C could induce allergy. We examined the protein with three other proteins to determine the potential allergenicity of Cry1C protein from genetically modified rice. Female Brown Norway (BN) rats received 0.1 mg peanut agglutinin (PNA), 1mg potato acid phosphatase (PAP), 1mg ovalbumin (OVA) or 5 mg purified Cry1C protein dissolved in 1 mL water by daily gavage for 42 days to test potential allergenicity. Ten days after the last gavage, rats were orally challenged with antigens, and physiologic and immunologic responses were studied. In contrast to sensitization with PNA, PAP and OVA Cry1C protein did not induce antigen-specific IgG2a in BN rats. Cytokine expression, serum IgE and histamine levels and the number of eosinophils and mast cells in the blood of Cry1C group rats were comparable to the control group rats, which were treated with water alone. As Cry1C did not show any allergenicity, we make the following conclusion that the protein could be safety used in rice or other plants.

Topics: Acid Phosphatase; Animals; Cytokines; Enzyme-Linked Immunosorbent Assay; Eosinophils; Escherichia coli; Female; Food Hypersensitivity; Food, Genetically Modified; Histamine; Immunoglobulin E; Immunoglobulin G; Mast Cells; Oryza; Ovalbumin; Peanut Agglutinin; Plant Proteins; Plants, Genetically Modified; Rats; Rats, Inbred BN; Real-Time Polymerase Chain Reaction

We examined whether maternal exposure to food antigens during lactation and maternal allergic status would affect the development of food allergy in offspring. OVA-sensitized or OVA-nonsensitized BALB/c female mice were exposed or unexposed to OVA during lactation. After weaning, their offspring were systemically sensitized twice with OVA and repeatedly given OVA by oral intubation. While 97.1% of the mice breastfed by OVA-nonsensitized and OVA-unexposed mothers developed allergic diarrhea, 59.7% of the mice breastfed by OVA-exposed nonallergic mothers during lactation and 24.6% of the mice breastfed by OVA-exposed allergic mothers during lactation developed food allergy. Furthermore, OVA was detected in breast-milk from OVA-exposed nonallergic mothers during lactation (4.6 ± 0.5 μg/mL). In addition, OVA-specific IgG1 titers were markedly increased in breast milk from allergic mothers (OVA-sensitized and OVA-unexposed mother: 11.0 ± 0.5, OVA-sensitized and OVA-exposed mother: 12.3 ± 0.3). Our results suggest that oral tolerance induced by breast milk-mediated transfer of dietary antigens along with their specific immunoglobulins to offspring leads to antigen-specific protection from food allergy.

Topics: Administration, Oral; Allergens; Animals; Animals, Newborn; Disease Models, Animal; Female; Food Hypersensitivity; Humans; Immune Tolerance; Immunity, Maternally-Acquired; Immunoglobulin E; Immunoglobulin G; Mice; Mice, Inbred BALB C; Milk; Ovalbumin

Anti-allergic efficacy of red ginseng (RG) and fermented red ginseng (FRG) was evaluated. RG or FRG were administered to ovalbumin (OVA)-sensitized mice for 8 weeks. Immunoglobulins (Igs), Th1/Th2 type cytokines, and β-lactoglobulin (BLG) in serum, and intestinal barrier-related molecules in jejunum were measured using enzyme-linked immunosorbent assay or reverse transcription-polymerase chain reaction. Mice sensitized with OVA increased serum IgG₁, IgE, OVA-IgG₁, and OVA-IgE. Both RG and FRG decreased serum IgE, OVA-IgE, and pro-inflammatory cytokines. Serum BLG, a marker of gut permeability, was significantly higher in sensitized animals and was decreased in mice fed RG or FRG. In addition, intestinal barrier-related markers such as MMCP-1, IL-4, TNF-α, COX-2, and iNOS mRNA expressions were decreased by RG or FRG. Our results suggest in vivo anti-allergic activities of RG or FRG, which are associated with the regulation of Th1/Th2 balance, intestinal inflammation and subsequent the suppression of IgE.

Topics: Administration, Oral; Animals; Anti-Allergic Agents; Cytokines; Dietary Proteins; Female; Fermentation; Food Hypersensitivity; Immunoglobulin E; Immunoglobulin G; Inflammation; Inflammation Mediators; Intestinal Diseases; Intestinal Mucosa; Jejunum; Lactoglobulins; Mice; Mice, Inbred BALB C; Ovalbumin; Panax; Permeability; Phytotherapy; Plant Extracts; RNA, Messenger; Th1-Th2 Balance

The aim of the study was to investigate the effect of living probiotics, probiotic DNA and the synthetic oligodeoxynucleotides containing CpG motifs (CpG-ODN) on both immune response and intestinal barrier function in ovalbumin-sensitized rat and the underlying mechanisms. Brown-Norway rats were orally sensitized with ovalbumin, and living probiotics, probiotic DNA extraction, synthetic CpG-ODN or non-CpG ODN control was administered. In the living probiotics, probiotic DNA and CpG-ODN groups, the allergic response was significantly inhibited, the Th1/Th2 cytokine balance was shifted away from Th2 side, the percentage of CD4(+) CD25(+high) Treg cells was increased, and the intestinal barrier function was improved. The levels of toll-like receptor (TLR) 9 mRNA and nuclear factor (NF)-κB activity, as well as the IκB-α phosphorylation (p-IκB-α) was significantly increased in these three intervention groups compared with the OVA-positive group, whereas no such effects were found in the non-CpG ODN control group. These data show that the probiotic genomic DNA and the synthetic CpG-ODN was comparable with living probiotics in preventing food allergic response by immune modulation and intestinal barrier function enhancement, and the activation of TLR9/NF-κB signal pathway might be involved in this process.

Topics: Animals; Cytokines; DNA, Bacterial; Female; Food Hypersensitivity; Immunologic Factors; NF-kappa B; Oligodeoxyribonucleotides; Ovalbumin; Probiotics; Rats; Rats, Inbred BN; T-Lymphocytes, Regulatory; Toll-Like Receptor 9

The egg protein ovalbumin (OVA) belongs to six most frequent food allergens. We investigated how thermal processing influences its ability to induce allergic symptoms and immune responses in mouse model of food allergy.. Effect of increased temperature (70°C and 95°C) on OVA secondary structure was characterized by circular dichroism and by the kinetics of pepsin digestion with subsequent HPLC. BALB/c mice were sensitized intraperitoneally and challenged with repeated gavages of OVA or OVA heated to 70°C (h-OVA). Levels of allergen-specific serum antibodies were determined by ELISA (IgA and IgGs) or by β-hexosaminidase release test (IgE). Specific activities of digestive enzymes were determined in brush border membrane vesicles of jejunal enterocytes. Cytokine production and changes in regulatory T cells in mesenteric lymph nodes and spleen were assessed by ELISA and FACS. Heating of OVA to 70°C caused mild irreversible changes in secondary structure compared to boiling to 95°C (b-OVA), but both OVA treatments led to markedly different digestion kinetics and Tregs induction ability in vitro, compared to native OVA. Heating of OVA significantly decreased clinical symptoms (allergic diarrhea) and immune allergic response on the level of IgE, IL-4, IL-5, IL-13. Furthermore, h-OVA induced lower activities of serum mast cell protease-1 and enterocyte brush border membrane alkaline phosphatase as compared to native OVA. On the other hand h-OVA stimulated higher IgG2a in sera and IFN-γ secretion by splenocytes.. Minor irreversible changes in OVA secondary structure caused by thermal processing changes both its digestion and antigenic epitopes formation, which leads to activation of different T cell subpopulations, induces shift towards Th1 response and ultimately reduces its allergenicity.

Topics: Animals; Antigen Presentation; Cytokines; Disease Models, Animal; Female; Food Hypersensitivity; Hot Temperature; Immunoglobulin E; Mice; Ovalbumin; Protein Structure, Secondary; T-Lymphocytes, Regulatory

When sensitized epicutaneously and challenged orally with ovalbumin, Balb/c mice develop allergen-induced diarrhea. As mast cells play important roles in diarrhea, we studied whether allergic diarrhea could be alleviated with imatinib mesylate.. Balb/c mice were sensitized and challenged with ovalbumin and treated orally with imatinib. Cytokine mRNA expressions were determined with quantitative RT-PCR and numbers of small intestinal mast cells determined by staining for chloroacetate esterase and mucosal mast cell protease-1. Immunofluorescence staining was used to assess the intestinal CCL1 expression.. Ovalbumin-sensitized and challenged Balb/c mice developed diarrhea, which was associated with increased number of mast cells and expression of interleukin (IL)-4 and -13, and chemokines CCL1 and CCL17 in the small intestine. Treatment with imatinib reduced the incidence of diarrhea, inhibited the development of mastocytosis and jejunal mRNA expression of IL-13, CCL1, CCL17 and CCL22. Mast cell-deficient W/W(-V) mice, and surprisingly, also their mast cell-competent control (+/+) littermates failed to develop diarrhea as a response to ovalbumin. This strain-dependent difference was associated with the inability of +/+ and W/W(-V) mice to increase the number of intestinal mast cells and expression of IL-4, IL-13, CCL1 and CCL17 after ovalbumin challenge.. Development of allergic diarrhea is associated with the ability of mice to develop intestinal mastocytosis. Imatinib inhibited the development of intestinal mastocytosis, reduced the incidence of diarrhea, and reduced the expression of IL-13, CCL1, and CCL17. Targeting intestinal mast cells could be a feasible approach to treat allergic diarrhea.

Topics: Allergens; Animals; Benzamides; Diarrhea; Disease Models, Animal; Female; Food Hypersensitivity; Imatinib Mesylate; Intestines; Mast Cells; Mice; Ovalbumin; Piperazines; Protein Kinase Inhibitors; Pyrimidines; Reverse Transcriptase Polymerase Chain Reaction

The incidence of food hypersensitivity and food allergies is on the rise and new treatment approaches are needed. We investigated whether N. sativa, one of its components, thymoquinone, or synthetic opioid receptor (OR)-agonists can alleviate food allergy. Hence, ovalbumin (OVA)-sensitized BALB/c-mice were pre-treated either with a hexanic N. sativa seed extract, thymoquinone, kappa-(U50'4889) or mu-OR-agonists (DAMGO) and subsequently challenged intra-gastrically with OVA. All 4 treatments significantly decreased clinical scores of OVA-induced diarrhea. N. sativa seed extract, thymoquinone, and U50'488 also decreased intestinal mast cell numbers and plasma mouse mast cell protease-1 (MMCP-1). DAMGO, in contrast, had no effect on mast cell parameters but decreased IFNγ, IL-4, IL-5, and IL-10 concentration after ex vivo re-stimulation of mesenteric lymphocytes. The effects on allergy symptoms were reversible by OR-antagonist pre-treatment, whereas most of the effects on immunological parameter were not. We demonstrate that N. sativa seed extract significantly improves symptoms and immune parameters in murine OVA-induced allergic diarrhea; this effect is at least partially mediated by thymoquinone. ORs may also be involved and could be a new target for intestinal allergy symptom alleviation. N. sativa seed extract seems to be a promising candidate for nutritional interventions in humans with food allergy.

Topics: Animals; Benzoquinones; Biomarkers; Chymases; Diarrhea; Food Hypersensitivity; Ligands; Male; Mice; Mice, Inbred BALB C; Nigella sativa; Ovalbumin; Phytotherapy; Plant Extracts; Receptors, Opioid; Receptors, Opioid, kappa; Receptors, Opioid, mu; Seeds

Because few curative treatments are available for food allergy, we investigated the therapeutic potential of rapamycin, a mammalian target of rapamycin (mTOR) inhibitor, on mouse food allergy.. The preventive and therapeutic effects of oral rapamycin on anaphylactic symptoms induced by oral ovalbumin (OVA) challenge in food allergy mice were investigated. Mast cell functions in response to rapamycin were also measured in the passive systemic anaphylaxis model and bone marrow-derived mast cells (BMMCs).. Daily rapamycin from the first challenge (preventive protocol) attenuated food allergy symptoms including diarrhea, anaphylactic reactions, and hypothermia in mice. The treatment decreased the challenge-induced increases in mouse mast cell protease-1 in serum and mast cell numbers in the intestine. Notably, the mice that already showed food allergy symptoms by previous challenges recovered from the disease with daily administration of rapamycin (therapeutic protocol). Anti-OVA IgG1 and IgE levels in serum, as well as IFN-γ, IL-4, IL-13, IL-9, IL-10, and IL-17 secretion from splenocytes, were decreased by the treatments. In contrast, a single dose of rapamycin failed to affect passive systemic anaphylaxis. Spontaneous and IL-9-dependent survival and IgE-induced IL-13 secretion, but not degranulation, of BMMCs were reduced by rapamycin.. Our data show that mouse food allergy was attenuated by rapamycin through an immunosuppressive effect and inhibition of intestinal mast cell hyperplasia. Inhibition of the IL-9 production-mast cell survival axis is one of the mechanisms of the therapeutic effect of rapamycin. Rapamycin and other mTOR inhibitors might be good candidates for therapeutic drugs for food allergy.

Topics: Administration, Oral; Anaphylaxis; Animals; Food Hypersensitivity; Immunosuppressive Agents; Male; Mast Cells; Mice; Mice, Inbred BALB C; Ovalbumin; Sirolimus; Treatment Outcome

To investigate immune state in lung of BALB/c mice with ovalbumin (OVA) allergy and the effects of fulvotomentoside (Ful) on lungs of the mice and provide some clues for the mechanism that patients with food allergies were prone to asthma and observe the effects of the treatment with traditional Chinese medicine.. Ninety-six female BALB/c mice were randomly divided into 6 groups. Mice in group 1 and group 2 were sensitized intraperitoneally and challenged intragastrically with OVA and were exposed to phosphate buffer solution and OVA respectively by nebulized inhalation. Mice in group 3 and group 4 were treated with Ful, other processes were the same as the mice in group 1 and group 2, respectively. Mice in group 5 were not challenged intragastrically with OVA and other processes were the same as the mice in group 2. Group 6 was the control group. The number of total leukocytes and cell classification in bronchoalveolar lavage (BALF) were counted, and inflammatory characteristic of lung was scored by staining with hematoxylin and eosin. The protein expressions of transforming growth factor (TGF-β1), interleukin-6 (IL-6), interleukin-17 (IL-17A) in lung of the mice were detected by immunohistochemical method. The activation of neutrophils in lung was assayed by the level of myeloroxidase (MPO).. There was no inflammatory cells infiltration in lung of the mice in group 1. Compared with group 6, numbers of total leukocytes and erythrocytes as well as the percentage of neutrophils and lymphocytes were increased in group 2. Inflammatory score and protein expressions of TGF-β1 [(75 437 ± 3 638) vs. (6 118 ± 1 978)], IL-6 [(121 650 ± 25 389) vs. (15 726 ± 9 360)], IL-17A [(252 105 ± 31 651)vs. (72 644 ± 12 285)] in lung were increased, too. Inflammatory score and TGF-β1 (11 054 ± 1 468), IL-6 (50 877 ± 11 744), IL-17A (137 864 ± 28 986) expressions in group 5 were lower than those in group 2. Eosinophils infiltration was significant in group 5. After the treatment with Ful, TGF-β1 expression did not change and IL-6, IL-17A expressions were decreased in lung of the mice that inhaled OVA. It was not enough for Ful to relieve the neutrophil aggregation and improve inflammatory reaction in lung.. The expressions of TGF-β1, IL-6, IL-17A in lung of the mice with OVA allergy were increased markedly after they inhaled specific antigen, which caused serious inflammation that was induced by neutrophil infiltration in lung. Ful could decrease the expressions of IL-6, IL-17A to some extent, but it was not enough to improve pathologic state in lung.

Topics: Administration, Inhalation; Animals; Bronchoalveolar Lavage Fluid; Disease Models, Animal; Drugs, Chinese Herbal; Female; Food Hypersensitivity; Immunohistochemistry; Inflammation; Interleukin-17; Interleukin-6; Lung Diseases; Mice; Mice, Inbred BALB C; Neutrophils; Oleanolic Acid; Ovalbumin; Saponins; Transforming Growth Factor beta1

Resveratrol is a bioactive polyphenol enriched in red wine that exhibits many beneficial health effects via multiple mechanisms. However, it is unclear whether resveratrol is beneficial for the prevention of food allergy. This study investigated whether resveratrol inhibited the development of food allergy by using a mouse model of the disease.. Mice fed standard diet or standard diet plus resveratrol were sensitized by intragastric administration of ovalbumin (OVA) and mucosal adjuvant cholera toxin (CT). Several manifestations of food allergy were then compared between the mice. The effects of resveratrol on T cells or dendritic cells were also examined by using splenocytes from OVA-specific T cell-receptor (TCR) transgenic DO11.10 mice or mouse bone marrow-derived dendritic cells (BMDCs) in vitro. We found that mice fed resveratrol showed reduced OVA-specific serum IgE production, anaphylactic reaction, and OVA-induced IL-13 and IFN-ã production from the mesenteric lymph nodes (MLNs) and spleens in comparison to the control mice, following oral sensitization with OVA plus CT. In addition, resveratrol inhibited OVA plus CT-induced IL-4, IL-13, and IFN-ã production in splenocytes from DO11.10 mice associated with inhibition of GATA-3 and T-bet expression. Furthermore, resveratrol suppressed the OVA plus CT-induced CD25 expression and IL-2 production in DO11.10 mice-splenocytes in association with decreases in CD80 and CD86 expression levels. Finally, resveratrol suppressed CT-induced cAMP elevation in association with decreases in CD80 and CD86 expression levels in BMDCs.. Ingestion of resveratrol prevented the development of a food allergy model in mice. Given the in vitro findings, resveratrol might do so by inhibiting DC maturation and subsequent early T cell activation and differentiation via downregulation of CT-induced cAMP activation in mice. These results suggest that resveratrol may have potential for prophylaxis against food allergy.

Topics: Animals; Antigens, CD; Antioxidants; Cholera Toxin; Cyclic AMP; Dendritic Cells; Female; Food Hypersensitivity; GATA3 Transcription Factor; Gene Expression; Immunoglobulin E; Interferon-alpha; Interleukin-13; Interleukin-4; Lymph Nodes; Mice; Ovalbumin; Resveratrol; Signal Transduction; Spleen; Stilbenes; T-Box Domain Proteins; T-Lymphocytes

The epitopes for OVA323-339-specific CD4⁺T cells from OVA23-3 food allergy model and DO11.10 tolerant induction model mice were analyzed. We found that OVA23-3 CD4⁺T cells recognized the N-terminal region, showing strong proliferation and the Th2-phenotype, and that DO11.10 CD4⁺T cells recognized the C-terminal region, showing milder proliferation and a Th1-skewed response. These differences may regulate the responses of those mice to OVA-feeding, inflammation and tolerance.

Topics: Amino Acid Sequence; Animals; CD4-Positive T-Lymphocytes; Cell Differentiation; Epitopes; Food Hypersensitivity; Mice; Molecular Sequence Data; Ovalbumin; Peptide Fragments; Phenotype; Th1 Cells; Th2 Cells

To establish a BN rat model for testing food allergenicity.. BN rats of both sex and aged 4 and 8 weeks were exposed to different doses of ovalbumin (OVA) (1.00 mg, 0.10 mg and 0.01 mg) by intraperitoneal injection on the 1st, 5th and 10th day of the study and observed for 35 days. Blood samples were taken on the 28th and 35th day of the study from orbital plexus and serum specific IgE (OVA sIgE) were determined by ELISA.. Compared with the control group, the concentrations of OVA sIgE of 8-week female BN rats on the 28th and 35th day were significantly increased when 1.00 mg and 0.10 mg OVA was applied (P < 0.05). The concentrations of OVA sIgE of 8-week male BN rats on the 28th day were significantly higher than that in the control group when 0.10 mg and 0.01 mg OVA was applied (P < 0.05). And the concentrations of OVA sIgE of 8-week male BN rats on the 35th day were significantly higher than that in the control group when 1.00 mg OVA was applied (P < 0.05). The concentrations of OVA sIgE of 4-week male BN rats on the 35th day were significantly higher than the control group when 1.00 mg OVA was applied (P < 0.05).. Female rats were more sensitive than male rats to different doses of OVA administered intraperitoneally; 8-week rats were more sensitive than 4-week rats; the better dose of OVA for sensitizing BN rats was 0.10 mg or 1.00 mg. Therefore, an ideally sensitized animal model could be established by administering 8-week female BN rats with 0.10 mg or 1.00 mg OVA intraperitoneally in 35 days later.

Topics: Administration, Oral; Allergens; Animals; Disease Models, Animal; Enzyme-Linked Immunosorbent Assay; Female; Food Hypersensitivity; Humans; Immunoglobulin E; Male; Ovalbumin; Rats; Rats, Inbred BN

Food allergy is a serious health concern among infants and young children. Although immunological mechanism of food allergy is well documented, the molecular mechanism(s) involved in food allergen sensitization have not been well characterized. Therefore, the present study analyzed the mesenteric lymph node (MLN) transcriptome profiles of BALB/c mice in response to three common food allergens.. Microarray analysis identified a total of 1361, 533 and 488 differentially expressed genes in response to β-lactoglobulin (BLG) from cow's milk, ovalbumin (OVA) from hen's egg white and peanut agglutinin (PNA) sensitizations, respectively (p < 0.05). A total of 150 genes were commonly expressed in all antigen sensitized groups. The expression of seven representative genes from microarray experiment was validated by real-time RT-PCR. All allergens induced significant ear swelling and serum IgG1 concentrations, whereas IgE concentrations were increased in BLG- and PNA-treated mice (p < 0.05). Treatment with OVA and PNA significantly induced plasma histamine concentrations (p < 0.05). The PCA demonstrated the presence of allergen-specific IgE in the serum of previously sensitized and challenged mice.. Immunological profiles indicate that the allergen dosages used are sufficient to sensitize the BALB/c mice and to conduct transcriptome profiling. Microarray studies identified several differentially expressed genes in the sensitization phase of the food allergy. These findings will help to better understand the underlying molecular mechanism(s) of food allergen sensitizations and may be useful in identifying the potential biomarkers of food allergy.

Topics: Allergens; Animals; Disease Models, Animal; Food Hypersensitivity; Gene Expression Profiling; Histamine; Immunoglobulin E; Immunoglobulin G; Lactoglobulins; Lymph Nodes; Mice; Mice, Inbred BALB C; Ovalbumin; Peanut Agglutinin

Tolerance to food antigen manifests in the absence and/or suppression of antigen-specific immune responses locally in the gut but also systemically, a phenomenon known as oral tolerance. Oral tolerance is thought to originate in the gut-draining lymph nodes, which support the generation of FoxP3(+) regulatory T (Treg) cells. Here we use several mouse models to show that Treg cells, after their generation in lymph nodes, need to home to the gut to undergo local expansion to install oral tolerance. Proliferation of Treg cells in the intestine and production of interleukin-10 by gut-resident macrophages was blunted in mice deficient in the chemokine (C-X3-C motif) receptor 1 (CX3CR1). We propose a model of stepwise oral tolerance induction comprising the generation of Treg cells in the gut-draining lymph nodes, followed by migration into the gut and subsequent expansion of Treg cells driven by intestinal macrophages.

Topics: Adoptive Transfer; Animals; Cell Division; Chemotaxis, Leukocyte; CX3C Chemokine Receptor 1; Diarrhea; Food Hypersensitivity; Forkhead Transcription Factors; Immune Tolerance; Immunity, Mucosal; Interleukin-10; Lymph Nodes; Macrophages; Mice; Mice, Inbred BALB C; Mice, Transgenic; Mucous Membrane; Ovalbumin; Receptors, Chemokine; Receptors, Lymphocyte Homing; T-Lymphocytes, Regulatory

The mechanism of intestinal immune inflammation, such as food allergy, remains to be further understood. The present study aims to investigate the role of the vagal nerve in the pathogenesis of skewed T-helper 2 (Th2) responses in the intestine.. The expression of the immunoglobulin E (IgE) receptor on the vagus nerve in the mouse intestine was observed by immunohistochemistry. Vagus ganglion neurons (VGN) were isolated from mice and cultured in vitro. The IgE receptor/IgE complex on vagus neurons was examined by immune precipitation assay. A food allergy mouse model was developed; the effect of the partial removal of the vagal nerve (PRVn) via surgery or administration with anticholinergic agents on the suppression of Th2 inflammation was evaluated.. The high-affinity IgE receptor was detected on the intestinal vagus nerve. An increase in the expression of the IgE receptor on the vagus nerve was observed in the intestines of mice with intestinal immune inflammation. Isolated mouse VGN express IgE receptor I, which could form complexes with IgE. Re-exposure to specific antigens activated the sensitized VGN, manifesting the release of transmitter glutamate that could activate dendritic cells by increasing the expression of CD80 and major compatibility complex class II and suppressing interleukin-12. The PRVn suppressed Th2 inflammation in the intestine.. The intestinal vagus nerve in mice expresses a high-affinity IgE receptor. An antigen-specific immune response can activate the vagus nerve in the intestine and induces the release of transmitters to modulate dendritic cell phenotypes that facilitate the development of skewed Th2 polarization in the intestine.

Topics: Analysis of Variance; Animals; B7-1 Antigen; Cells, Cultured; Cholinergic Antagonists; Dendritic Cells; Disease Models, Animal; Enzyme-Linked Immunosorbent Assay; Food Hypersensitivity; Ganglia, Parasympathetic; Glutamic Acid; Histocompatibility Antigens Class II; Immunoglobulin E; Immunohistochemistry; Inflammation; Interleukin-12; Intestines; Male; Mice; Mice, Inbred BALB C; Ovalbumin; Receptors, IgE; Th2 Cells; Vagotomy; Vagus Nerve

Patients with atopic dermatitis (AD) often suffer from food allergy and develop flares upon skin contact with food allergens. However, it is unclear whether T cells sensitized to allergens in the gut promote this skin inflammation. To address this question, we orally immunized WT mice and mice lacking the skin-homing chemokine receptor Ccr4 (Ccr4-/- mice) with OVA and then challenged them epicutaneously with antigen. Allergic skin inflammation developed in the WT mice but not in the mutants and was characterized by epidermal thickening, dermal infiltration by eosinophils and CD4+ T cells, and upregulation of Th2 cytokines. T cells purified from mesenteric lymph nodes (MLNs) of orally immunized WT mice transferred allergic skin inflammation to naive recipients cutaneously challenged with antigen, but this effect was lost in T cells purified from Ccr4-/- mice. In addition, the ability of adoptively transferred OVA-activated T cells to home to the skin following cutaneous OVA challenge was ablated in mice that lacked lymph nodes. These results indicate that cutaneous exposure to food antigens can reprogram gut-homing effector T cells in LNs to express skin-homing receptors, eliciting skin lesions upon food allergen contact in orally sensitized AD patients.

Topics: Administration, Cutaneous; Administration, Oral; Adoptive Transfer; Allergens; Animals; Chemotaxis, Leukocyte; Cholera Toxin; Dermatitis, Allergic Contact; Food Hypersensitivity; Immunization; Integrins; Mice; Mice, Inbred BALB C; Mice, Inbred C57BL; Ovalbumin; Receptors, CCR4; Receptors, Fibroblast Growth Factor; Receptors, Lymphocyte Homing; Sialoglycoproteins; Skin; Specific Pathogen-Free Organisms; T-Lymphocyte Subsets

Modulating early immune response by application of bacteria and their by-products has been suggested as a preventive strategy against the development of allergic diseases. In light of this, the aim of the study was to test the effects of oral administration of bacterial lysates (BL) in a rat model of food allergy.. BL or PBS were administered orally to neonatal Brown Norway rats up to an age of 42 days. Additionally, animals were sensitized 3 times (days 35, 40 and 45) intraperitoneally with ovalbumin (OVA). On days 60 and 61, rats were locally challenged with OVA by gavage feeding.. Detection of increased allergen-specific Ig serum levels and proliferative responses of spleen mononuclear cells confirmed systemic sensitization. In serum of animals that received BL in addition to OVA sensitization, the levels of allergen-specific IgE and IgG were significantly reduced compared to animals which were not exposed to BL. Allergen-stimulated lymphocytes from spleen and mesenteric lymph nodes of BL-treated animals showed a significantly elevated cytokine production of IL-10. To assess local functional changes of the intestinal barrier we measured the intestinal permeability, which was increased in OVA-sensitized and challenged animals compared to nonsensitized controls, yet significantly reduced in sensitized animals which received BL.. These data suggest that local administration of BL (pathogen-associated molecular patterns) in the intestine exhibits immuno-modulating effects. Furthermore, pathophysiological features of food allergy, such as the loss of gut mucosal integrity, might be reduced by the treatment with BL.

Topics: Administration, Oral; Animals; Animals, Newborn; Cell Extracts; Enzyme-Linked Immunosorbent Assay; Food Hypersensitivity; Immunoglobulin E; Immunoglobulin G; Intestinal Diseases; Ovalbumin; Rats; Specific Pathogen-Free Organisms

To investigate the consequences of food allergy in adipose tissue and metabolism, we used a murine model in which mice have been sensitized subcutaneously with ovalbumin and further received antigen-containing diet. Allergic mice presented a significant weight loss 7 days after oral challenge with a concomitant decrease in epididymal adipose tissue mass. This decrease was associated with increased lipolysis and local inflammation. In adipose tissue of allergic mice there were increased leukocyte rolling and adhesion in the microvasculature, increased number of leukocytes in the tissue, especially macrophages (F4/80(+) cells) and increased pro-inflammatory cytokines levels, including TNF-α, IL-6 and CCL2. In addition, we observed low serum concentrations of triglyceride, glucose, total cholesterol and free fatty acids in the allergic mice. Our results suggest that the induction of food allergy in mice leads to adipose tissue inflammation and systemic metabolic alterations that contribute to the weight loss observed.

Topics: Adipose Tissue; Animals; Blood Glucose; Cell Adhesion; Chemokines; Cholesterol; Cytokines; Epididymis; Fatty Acids, Nonesterified; Food Hypersensitivity; Inflammation; Leukocyte Rolling; Lipolysis; Macrophages; Male; Mast Cells; Mice; Mice, Inbred BALB C; Ovalbumin; Triglycerides; Weight Loss

Recent studies have revealed that various neurotransmitters regulate the immune system via their receptors expressed on the immune cells. Calcitonin gene-related peptide (CGRP), a sensory nerve C-fiber neuropeptide, is also known to have the ability to modulate the functions of immune cells in vitro. However, the contribution of CGRP to the immune regulation in vivo remains to be fully elucidated. Here we report that mice deficient in receptor activity-modifying protein 1 (RAMP1), which is a subunit of the CGRP receptor, showed a significantly lower incidence of diarrhea compared with wild-type (WT) mice in the ovalbumin (OVA)-induced food allergic model. Serum OVA-specific IgE levels and the differentiation of T helper cells was comparable in WT mice and RAMP1-deficient mice. Moreover, there were no significant differences between recruitment and degranulation of mast cells in the small intestine of these mice. In contrast, significantly diminished intestinal peristalsis was observed by the allergy induction in RAMP1-deficient mice compared with WT mice. These results suggest that this suppression of allergic diarrhea is due to the diminished intestinal peristalsis in RAMP1-deficient mice.

Topics: Animals; Diarrhea; Food Hypersensitivity; Intestines; Mice; Mice, Mutant Strains; Ovalbumin; Peristalsis; Receptor Activity-Modifying Protein 1

Oral immunotherapy (OIT) involving continuous oral administration of allergenic foods has gained attention as a therapy for food allergies. To study the influence of oral administration of allergenic foods on gastrointestinal symptoms including inflammation, we established a mouse model of food-induced intestinal allergy.. BALB/c mice were fed an egg white (EW) diet containing ovalbumin (OVA, a major EW allergen) after intraperitoneal sensitisation with OVA and Alum. The mice on the EW diet for one wk presented gastrointestinal symptoms (i.e. weight loss and soft stools) and inflammation in the small intestines (i.e. duodenum, jejunum and ileum). Further continuous EW diet resolved the weight loss but not the soft stools. Splenic CD4(+) T-cells of EW diet-fed mice on the continuous diet showed less proliferation and cytokine production compared with those of control mice, suggesting tolerance induction by the diet. The continuous EW diet reduced levels of OVA-specific IgE antibodies, but significantly aggravated the inflammation in the jejunum.. Our mouse model would be useful to investigate inflammatory and regulatory mechanisms in food-induced intestinal allergies. Our results suggest potential gastrointestinal inflammation in patients undergoing OIT as continuous administration of allergenic foods, even though the therapy may induce clinical tolerance.

Topics: Administration, Oral; Animals; CD4-Positive T-Lymphocytes; Cytokines; Disease Models, Animal; Egg White; Female; Food; Food Hypersensitivity; Gastroenteritis; Immune Tolerance; Immunoglobulin E; Interleukin-10; Intestine, Small; Mice; Mice, Inbred BALB C; Ovalbumin; Spleen

This study focuses on the effect of heating and Maillard reaction (MR) on the in vitro digestibility and rabbit IgG- and human IgE-binding properties of ovalbumin (OVA) and ovomucoid (OM) to estimate the impact of processing on their allergenicity. With the human sera studied, heat treatment significantly reduced IgE binding to both OVA and OM, whereas MR reduced the IgE binding to OVA but increased IgE binding to OM. In contrast, heat treatment significantly favored OVA digestibility but glycation impaired it, and these treatments did not affect the digestibility of OM. The changes observed in the digestibility affected the immunogenicity of the digests accordingly, so that the higher the digestibility, the lower the antibody binding. Heat treatment and glycation by MR showed an influence on the potential allergenicity of the main egg white proteins that could be related to their resistance to denaturation and digestive enzymes.

Topics: Allergens; Animals; Digestion; Food Hypersensitivity; Glycosylation; Hot Temperature; Humans; Immunoglobulin E; Maillard Reaction; Ovalbumin; Ovomucin; Rabbits

Food allergy has reached an epidemic level in westernized countries and although central mechanisms have been described, the variability associated with genetic diversity underscores the still unresolved complexity of these disorders.. To develop models of food allergy and oral tolerance, both strictly induced by the intestinal route, and to compare antigen-specific responses.. BALB/c mice were mucosally sensitized to ovalbumin (OVA) in the presence of the mucosal adjuvant cholera toxin, or tolerized by intra-gastric administrations of OVA alone. Antibody titres and cytokines were determined by ELISA, and allergic status was determined through several physiologic parameters including decline in temperature, diarrhoea, mast cell degranulation and intestinal permeability.. OVA-specific antibodies (IgE, IgGs and IgA in serum and feces) were produced in sensitized mice exclusively. Upon intra-gastric challenge with OVA, sensitized mice developed anaphylactic reactions associated with a decline of temperature, diarrhoea, degranulation of mast cells, which were only moderately recruited in the small intestine, and increased intestinal permeability. Cytokines produced by immune cells from sensitized mice included T-helper type 2 cytokines (IL-5, IL-13), but also IL-10, IFN-gamma and IL-17. In contrast, all markers of allergy were totally absent in tolerized animals, and yet the latter were protected from subsequent sensitization, demonstrating that oral tolerance took place efficiently.. This work allows for the first time an appropriate comparison between sensitized and tolerized BALB/c mice towards OVA. It highlights important differences from other models of allergy, and thus questions some of the generally accepted notions of allergic reactions, such as the protective role of IFN-gamma, the importance of antigen-specific secretory IgA and the role of mucosal mast cells in intestinal anaphylaxis. In addition, it suggests that IL-17 might be an effector cytokine in food allergy. Finally, it demonstrates that intestinal permeability towards the allergen is increased during challenge.

Topics: Administration, Oral; Allergens; Animals; Antibodies; Antibody Specificity; Cell Degranulation; Cells, Cultured; Cytokines; Disease Models, Animal; Female; Food Hypersensitivity; Immune Tolerance; Immunoglobulin A, Secretory; Interleukin-17; Intestinal Mucosa; Lymphocytes; Mast Cells; Mice; Mice, Inbred BALB C; Ovalbumin

Our preliminary clinical trial showed that consumption of cooked rice of a Japanese common cultivar Yukihikari improved atopic dermatitis associated with a suspected rice allergy, although the underlying mechanisms remain unclear. We hypothesised that the ameliorating effect of Yukihikari on atopic dermatitis is associated with the gut microbiota. BALB/c mice were fed a synthetic diet supplemented with uncooked and polished white rice powder prepared from one of four different cultivars: Yukihikari, rice A (common rice), rice B (brewery rice) and rice C (waxy rice). Denaturing gradient gel electrophoresis of PCR-amplified 16S rRNA gene fragments showed that the composition of faecal microbiota was different between mice fed Yukihikari and those fed rice A. Analysis of the 16S rRNA clone library and species-specific real-time PCR showed that the abundance of Akkermansia muciniphila, a mucin degrader, tended to be lower in mice fed Yukihikari. The incidence of allergic diarrhoea induced by oral administration of ovalbumin in systemically immunised mice was lower in mice fed Yukihikari, albeit with no difference in serum antibodies specific to ovalbumin. In a separate experiment, serum antibody levels specific to orally administered ovalbumin were lower in mice fed Yukihikari. Additionally, the translocation of horseradish peroxidase in isolated segments of ileum and colon tended to be lower in mice fed Yukihikari, suggesting a reduction in gut permeability in mice fed Yukihikari. These data indicate that changes in the gut microbiota of mice fed Yukihikari could be advantageous in the prevention of food allergy.

Topics: Administration, Oral; Animal Feed; Animals; Dermatitis, Atopic; Diarrhea; Female; Food Hypersensitivity; Humans; Immunoglobulin E; Immunoglobulin G; Mice; Mice, Inbred BALB C; Oryza; Ovalbumin

CCL20 is a chemokine that regulates the homeostatic and inflammatory trafficking of leukocytes to the small intestine and regulates the development of the gastrointestinal lymphoid architecture. T cells expressing T helper cell (Th) 2 cytokines are critical for experimental food allergy, and we hypothesized that CCL20 is involved in the localization of these cells to the gut.. We evaluated the role of CCR6 in allergic diarrhea induced by sensitization and oral challenge with ovalbumin (OVA) using CCR6(+/+) and CCR6(-/-) mice.. CCR6(-/-) mice were protected from OVA-induced diarrhea but surprisingly were not impaired in mastocytosis or allergen-specific immunoglobulin E. CCR6(-/-) mice were also protected from T cell-mediated diarrhea induced by anti-CD3 antibody. Allergic diarrhea was associated with an increased expression of Th2 cytokines within the intestinal mucosa that was significantly reduced in CCR6(-/-) mice. Inhibition of lymphocyte homing by treatment with FTY720 did not impair allergic diarrhea, indicating that reactivation of T cells could occur locally within the small intestine. Finally, T-cell transfer studies demonstrated that CCR6 was required both on the transferred T cells and in the recipient mouse to manifest allergic disease in the gastrointestinal tract.. These studies highlight a mast cell- and immunoglobulin E-independent role for CCR6-bearing T cells in the pathogenesis of gastrointestinal allergic disease.

Topics: Adoptive Transfer; Animals; Antibodies; CD3 Complex; Cells, Cultured; Chemokine CCL20; Cholera Toxin; Diarrhea; Female; Fingolimod Hydrochloride; Food Hypersensitivity; Immunoglobulin E; Immunosuppressive Agents; Jejunum; Lymph Nodes; Mast Cells; Mice; Mice, Inbred BALB C; Mice, Mutant Strains; Ovalbumin; Propylene Glycols; Receptors, CCR6; Sphingosine; Th2 Cells

The Maillard reaction occurs between reducing sugars and proteins during thermal processing of foods. It produces chemically glycated proteins termed advanced glycation end products (AGEs). The glycation structures of AGEs are suggested to function as pathogenesis-related immune epitopes in food allergy.. This study aimed at defining the T-cell immunogenicity of food AGEs by using ovalbumin (OVA) as a model allergen.. AGE-OVA was prepared by means of thermal processing of OVA in the presence of glucose. Activation of OVA-specific CD4(+) T cells by AGE-OVA was evaluated in cocultures with bone marrow-derived murine myeloid dendritic cells (mDCs) as antigen-presenting cells. The uptake mechanisms of mDCs for AGE-OVA were investigated by using inhibitors of putative cell-surface receptors for AGEs, as well as mDCs deficient for these receptors.. Compared with the controls (native OVA and OVA thermally processed without glucose), AGE-OVA enhanced the activation of OVA-specific CD4(+) T cells on coculture with mDCs, indicating that the glycation of OVA enhanced the T-cell immunogenicity of the allergen. The mDC uptake of AGE-OVA was significantly higher than that of the controls. We identified scavenger receptor class A type I and II (SR-AI/II) as a mediator of the AGE-OVA uptake, whereas the receptor for AGEs and galectin-3 were not responsible. Importantly, the activation of OVA-specific CD4(+) T cells by AGE-OVA was attenuated on coculture with SR-AI/II-deficient mDCs.. SR-AI/II targets AGE-OVA to the MHC class II loading pathway in mDCs, leading to an enhanced CD4(+) T-cell activation. The Maillard reaction might thus play an important role in the T-cell immunogenicity of food allergens.

Topics: Allergens; Animals; Antigen Presentation; Bone Marrow Cells; CD4-Positive T-Lymphocytes; CD8-Positive T-Lymphocytes; Coculture Techniques; Dendritic Cells; Food Hypersensitivity; Glycation End Products, Advanced; Lymphocyte Activation; Maillard Reaction; Mice; Mice, Inbred BALB C; Mice, Inbred C57BL; Ovalbumin; Scavenger Receptors, Class A; T-Lymphocytes

Mucosal immunity protects the epithelial barrier by immune exclusion of foreign antigens and by anti-inflammatory tolerance mechanisms, but there is a continuing debate about the role of secretory immunoglobulins (SIgs), particularly SIgA, in the protection against allergy and other inflammatory diseases. Lack of secretory antibodies may cause immune dysfunction and affect mucosally induced (oral) tolerance against food antigens.. We used polymeric Ig receptor (pIgR) knockout (KO) mice, which cannot export SIgA or SIgM, to study oral tolerance induction by ovalbumin (OVA) feeding and for parenteral antigen sensitization in the same animal.. Remarkable systemic hyperreactivity was observed in pIgR KO mice, as 50% died after intradermal OVA challenge, which was not seen in similarly sensitized and challenged wild-type (WT) mice. Oral tolerance induced by OVA completely protected the sensitized pIgR KO mice against anaphylaxis and suppressed antibody levels (particularly IgG1) as well as delayed-type hypersensitivity (DTH) to OVA. Delayed-type hypersensitivity to a bystander antigen, human serum albumin, was also suppressed and T-cell proliferation against OVA in vitro was reduced in tolerized compared with non-tolerized pIgR KO mice. This effect was largely mediated by CD25+ T cells. Adoptive transfer of splenic putative regulatory T cells (CD4+ CD25+) obtained from OVA-fed pIgR KO mice to naïve WT mice mediated suppression of DTH against OVA after sensitization of the recipients.. Compensatory regulatory T-cell function becomes critical in pIgR-deficient mice to avoid the potentially catastrophic effects of systemic immune hyperreactivity, presumably resulting from defective secretory antibody-mediated immune exclusion of microbial components.

Topics: Adoptive Transfer; Animals; Enzyme-Linked Immunosorbent Assay; Female; Food Hypersensitivity; Hypersensitivity, Delayed; Immune Tolerance; Immunity, Mucosal; Male; Mice; Mice, Inbred C57BL; Mice, Knockout; Ovalbumin; Receptors, Polymeric Immunoglobulin; T-Lymphocytes, Regulatory

Advanced glycation endproducts (AGEs) of food proteins resulting from the Maillard reaction after cooking or heating may have particular importance in food allergy. The underlying immunological mechanisms are only poorly understood. The aim of the study was to examine the effects of AGE derived from the model food allergen ovalbumin (AGE-OVA) on dendritic cells (DCs), their immunostimulatory capacity and the T-cell response compared with regular OVA. For this purpose, human immature DCs were exposed to fluorescein isothiocyanate (FITC)-labelled AGE-OVA and FITC-labelled regular OVA and uptake was analysed by flow cytometry and fluorescence microscopy. Furthermore, autologous CD4(+) T-cell proliferation and cytokine production induced by mature DCs loaded with AGE-OVA were compared with those induced by mature DCs loaded with OVA. Finally, expression of the receptor for advanced glycation endproducts (RAGE) and activation of the transcription factor nuclear factor (NF)-kappaB by AGE were investigated. Internalization of FITC-AGE-OVA by immature DCs was significantly increased compared with FITC-OVA. Blocking the mannose receptor, macropinocytosis or the scavenger receptor strongly reduced uptake of both FITC-OVA and FITC-AGE-OVA. In a comparison of CD4(+) T cells co-cultured with AGE-OVA-loaded mature DCs versus those co-cultured with OVA-loaded mature DCs, AGE-OVA DCs were found to produce more interleukin (IL)-6 and to induce a stronger T helper type 2 (Th2) and a weaker Th1 cytokine response, while there was no difference in proliferation of CD4(+) T cells. The expression of RAGE was higher on immature DCs compared with mature DCs. AGE-OVA-exposed immature DCs showed a stronger expression of RAGE and activation of the transcription factor NF-kappaB compared with OVA-loaded immature DCs. Our data indicate that AGE-OVA may be more immunogenic/allergenic than regular OVA.

Topics: Allergens; Antibodies; Antigen Presentation; CD4-Positive T-Lymphocytes; Cell Proliferation; Cytokines; Dendritic Cells; Endocytosis; Food Hypersensitivity; Glycation End Products, Advanced; Humans; Interleukin-6; Lymphocyte Activation; Ovalbumin; Phosphorylation; Receptor for Advanced Glycation End Products; Receptors, Immunologic; Th1 Cells; Th2 Cells; Transcription Factor RelA

Allergen-specific immunotherapy, subcutaneous immunotherapy (SCIT) or oral, has been used for almost a century to redirect inappropriate immune responses in atopic patients. A new mode of administration through the intact skin [epicutaneous immunotherapy (EPIT)], using an original epicutaneous delivery system, may represent an alternative to these classical methods.. Proof of concept of efficacy of EPIT on intact skin in mice sensitized to aeroallergens or food allergens.. Mice were sensitized to pollen (n=18), house dust mite (HDM, n=24), ovalbumin (OVA, n=18) or peanut (n=18), and allocated to four groups: EPIT, SCIT, not treated (NT) and control. Specific Ig (sIg)E, sIgG1 and sIgG2a were monitored. After 8 weeks of treatment, plethysmography was performed after aerosol provocation with appropriate allergens.. At the highest doses of methacholine, pause enhancement (Penh) values were significantly decreased in the EPIT group vs. the sensitized NT groups (7.5 vs. 12.3 - pollen, 7.6 vs. 8.9 - HDM, 11.5 vs. 14.5 - OVA, 7.6 vs. 12.8 - peanut, respectively) (P<0.05). With all the allergens tested, Penh values were similar in SCIT, EPIT and control. IgG2a for pollen, HDM, OVA and peanuts were significantly increased in the EPIT group vs. NT: 0.97 vs. 0.42 microg/mL, 2.5 vs. 0.46 microg/mL, 0.39 vs. 0.05 microg/mL and 15.0 vs. 5.5 microg/mL, respectively (P<0.05). There were no significant differences between EPIT and SCIT groups. The IgE/IgG2a ratio decreased significantly in the EPIT group for the four allergens from 70 to 58 (pollen), 175 to 26 (HDM), 5433 to 120 (OVA) and 49 to 6 (peanut), respectively (P<0.05).. In mice sensitized to the four allergens tested, EPIT was as efficacious as SCIT, considered as the reference immunotherapy. These first results have to be confirmed by clinical studies.

Topics: Animals; Arachis; Desensitization, Immunologic; Disease Models, Animal; Food Hypersensitivity; Humans; Hypersensitivity, Immediate; Mice; Mice, Inbred BALB C; Ovalbumin; Peanut Hypersensitivity; Pollen; Pyroglyphidae; Respiratory Hypersensitivity; Skin; Treatment Outcome

Animals sensitized to allergens change their feeding behavior and avoid drinking the otherwise preferred sweetened solutions containing the allergens. This phenomenon, known as food aversion, appears to be mediated by allergen-specific IgE antibodies. Here we investigated food aversion in BALB/c and C57BL/6 mice, which differ in their allergic responses to the allergen ovalbumin as well as in their preference for sweet taste. BALB/c mice present higher levels of IgE and a natural lower preference for sweet flavors when compared to C57BL/6 mice. Specifically, we studied a conflicting situation in which animals simultaneously experienced the aversive contact with the allergen and the attractive sweet taste of increasing concentrations of sucrose. We found that BALB/c mice were more prone to develop food aversion than C57BL/6 mice and that this aversive behavior could be abolished in both strains by increasing the palatability of the solution containing the allergen. In both strains food aversion was positively correlated with the levels of allergen-specific IgE antibodies and inversely correlated with their preference for sucrose sweetened solutions.

Topics: Allergens; Amino Acids; Animals; Feeding Behavior; Food Hypersensitivity; Food Preferences; Immune Sera; Immunization, Passive; Immunoglobulin E; Male; Mice; Mice, Inbred BALB C; Mice, Inbred C57BL; Ovalbumin; Species Specificity; Sweetening Agents; Taste

The incidence of atopic disease has increased dramatically during recent decades and the potential immunoregulatory influence of the microbiota in these individuals is under investigation.. The aim of our study was to identify a bacterial strain that is protective in murine allergy models and to determine if microbial induction of T regulatory cells was associated with protection from allergic inflammation.. Three microbes (Bifidobacterium breve AH1205, B. longum AH1206 and Lactobacillus salivarius AH102) of human origin were fed to newborn, adult and germ-free animals. Induction of Foxp3(+) T regulatory cells was assessed by flow cytometry. Gene array analysis was performed on Peyer's patches. Strains were also examined for their protective effects in the ovalbumin (OVA) respiratory allergy model and the OVA-cholera toxin dietary allergy model.. Bifidobacterium longum AH1206 consumption resulted in increased numbers of Foxp3(+) T regulatory cells in infant, adult and germ-free animals. B. breve AH1205 induced Foxp3(+) T regulatory cell expansion only in infant mice while L. salivarius AH102 did not alter T regulatory cell numbers in any animal model tested. B. longum AH1206 reduced the Peyer's patch gene expression associated with antigen presentation, TLR signalling and cytokine production while increasing the expression of genes associated with retinoic acid metabolism. B. longum AH1206 protected against airway inflammation in OVA-sensitized animals and B. longum AH1206 blocked the induction of IgE to orally administered OVA. Neither B. breve AH1205 nor L. salivarius AH102 had a protective effect in either model.. Bacterial strain-specific induction of Foxp3(+) T regulatory cells in vivo is associated with protection from respiratory and oral allergy.

Topics: Administration, Oral; Animals; Animals, Newborn; Bifidobacterium; Cholera Toxin; Disease Models, Animal; Food Hypersensitivity; Forkhead Transcription Factors; Lactobacillus; Mice; Mice, Inbred BALB C; Ovalbumin; Probiotics; Respiratory Hypersensitivity; Species Specificity; T-Lymphocytes, Regulatory

To ascertain the role of IL-4 in aversion to antigen induced by food allergy, wild type and IL-4 deficient BALB/c mice were sensitized with ovalbumin and challenged orally with egg white. Sensitized wild type mice had increased production of IL-4 by spleen and mesenteric lymph node cells in vitro, higher levels of serum anti-ovalbumin IgE and IgG1, aversion to ingestion of the antigen and loss of body weight after continuous oral challenge. Intestinal changes in wild type sensitized mice included eosinophil infiltration and increased mucus production. The IL-4 deficiency impaired the development of food allergy and the aversion to antigen, suggesting the involvement of the antigen specific antibodies. When IL-4 deficient mice received serum from sensitized wild type donors, the aversion was restored. These results indicate that production of IL-4 and specific IgE/IgG1 antibodies correlate with aversion to antigen induced by food allergy in mice.

Topics: Animals; Body Weight; Chickens; Digestion; Female; Food Hypersensitivity; Immunoglobulin E; Immunoglobulin G; Interleukin-4; Mice; Mice, Inbred BALB C; Mice, Knockout; Ovalbumin

The immune system may be modulated with nutrition to prevent the development or to treat the symptoms of allergy. Among other foods, consumption of apples has been linked to reduced incidence of atopic dermatitis and respiratory allergy.. We evaluated the efficacy and mechanisms of a polyphenol-enriched apple extract in reducing symptoms of food allergy.. In a model of food allergy to ovalbumin (OVA), BALB/c mice were fed with an apple extract either during sensitization or just before the challenge. After the challenge, allergic symptoms were scored, OVA-specific serum immunoglobulins were determined by ELISA, cytokine production by mesenteric lymph node (MLN) cells was measured by a multiplex assay and gene expression profiles in the intestine were addressed using quantitative real-time PCR.. Consumption of the apple extract reduced symptoms of food allergy upon challenge. This was paralleled by reduced levels of intestinal mast cell protease, diminished cytokine secretion by MLN cells and reduced local intestinal mRNA expression of various T-helper type-2 associated and pro-inflammatory genes. Mechanistic studies suggested decrease of mediator release by effector cells and reduction of allergenicity by protein-polyphenol interaction as potential mechanisms responsible for protection.. Polyphenol-enriched apple extract can attenuate food allergy symptoms in sensitized mice via two distinct possible mechanisms.

Topics: Animals; Chlorogenic Acid; Cytokines; Disease Models, Animal; Flavonoids; Food Hypersensitivity; Gene Expression Profiling; Hypersensitivity, Immediate; Immunoglobulins; Intestinal Mucosa; Mice; Mice, Inbred BALB C; Ovalbumin; Plant Extracts; Tannins; Treatment Outcome

Oral tolerance (OT) is being studied with great interest because of its therapeutic potential in allergy and autoimmunity. In the present study, two mouse strains with extreme phenotypes of OT susceptibility (TS) or resistance (TR) to ovalbumin (OVA) were used to demonstrate whether the tr and ts genes, cumulated during 18 generations of bi-directional genetic selection, influence expression of immunobiological traits in naive or antigen-gavaged TR/TS mice. The difference in anti-OVA titres was 2048-fold between OVA-gavaged TS and TR mice. Tolerance susceptibility to OVA gavage in individuals from a (TS x TR)F(2) population was 24% high-susceptibility, 62% low-susceptibility and 14% non-tolerant. Different antigens, unrelated to OVA, were tested by gavage and TS mice were generally susceptible while TR mice were resistant. The stability of TS and TR phenotypes was not affected by the use of strict protocols of intraperitoneal immunization or feeding over 30 consecutive days. The levels of interleukin-2 (IL-2), IL-4, interferon-gamma and IL-10 cytokines evaluated in concanavalin A-stimulated spleen cells from naive mice and in OVA-stimulated spleen cells from OVA-gavaged mice were higher in TS mice. Interleukin-10 was up-regulated in OVA-gavaged TS mice and down-regulated in TR mice. In naive mice, the percentage of CD4(+) CD25(+) and CD4(+) Foxp3(+) spleen cells and IL-10 expression by CD4(+) cells was significantly higher in TS mice. These results indicate that regulation of IL-10 expression could be an important factor contributing to the mechanisms controlling OT susceptibility, and that the OT responses of TR and TS individuals strongly correlate with their innate potential to secrete this cytokine.

Topics: Adjuvants, Immunologic; Animals; Antibodies; Antibody Formation; Antigens; B-Lymphocytes; CD4-Positive T-Lymphocytes; Cell Count; Cell Proliferation; Concanavalin A; Crosses, Genetic; Cytokines; Female; Food Hypersensitivity; Forkhead Transcription Factors; Genes, Dominant; Immune Tolerance; Immunity, Humoral; Immunoglobulin G; Interferon-gamma; Interleukin-10; Interleukin-2; Interleukin-2 Receptor alpha Subunit; Interleukin-4; Lipopolysaccharides; Lymphocyte Activation; Male; Mice; Mice, Inbred Strains; Ovalbumin; Phenotype; Spleen; T-Lymphocytes; T-Lymphocytes, Regulatory; Vaccination

Type-I allergic disorders and particularly food hypersensitivities are becoming increasingly common worldwide. This study investigated whether dietary enrichment with carotenoids inhibited oral sensitization to an antigen and the development of food allergies. The effects of a diet high in carotenoids were investigated in B10A mice that were orally sensitized to ovalbumin (OVA). The serum titers of OVA-specific immunoglobulin E (IgE), IgG1, and IgG2a were inhibited in mice fed ad libitum on a diet high in alpha- or beta-carotene compared to the control mice when orally sensitized to OVA. High alpha- and beta-carotene diets inhibited the immediate reduction in body temperature and rise in serum histamine associated with active systemic anaphylaxis in OVA-sensitized B10A mice. After re-stimulation with OVA in vitro, the production of T-helper 2-type cytokines by splenocytes from mice fed a diet high in carotenoids was lower than in control mice. Furthermore, the proportion of CD4(+) CD103(+) T cells in Peyer's patches of mice fed a carotenoid-rich diet was significantly lower than in control mice. These results suggest that an increased oral intake of carotenoids inhibits OVA-specific IgE and IgG1 production and antigen-induced anaphylactic responses by inhibiting specific T-cell activation in the mucosal immune system. A diet high in carotenoids might therefore prevent the development of food allergies.

Topics: Animals; Carotenoids; Cytokines; Disease Models, Animal; Female; Food Hypersensitivity; Humans; Immunoglobulin E; Immunoglobulin G; Mice; Ovalbumin; T-Lymphocytes

It was hypothesized that the suppressive effect of diosgenin (1) on the intestinal T helper (Th)2 responses is associated with an enhancement of the regulatory T-cell immunity. Ovalbumin (OVA)-sensitized BALB/c mice were gavaged daily with 1 and received repeatedly oral OVA challenges to induce intestinal allergic responses. The expression of Th2- and Treg-related cytokines and transcription factors was examined by immunohistochemical staining and RT-PCR. Administration of 1 markedly attenuated the intestinal expression of interleukin (IL)-4 and GATA3. In addition, administration of 1 reversed the diminished density of intestinal Foxp3(+) cells induced by OVA oral challenges and enhanced the expression of IL-10 by Foxp3(+) cells markedly. These results suggest that the suppressive effect of 1 on allergen-induced intestinal Th2 responses is closely associated with an up-regulation of the regulatory T-cell immunity in the inflammatory site.

Topics: Administration, Oral; Allergens; Animals; Dioscorea; Diosgenin; Food Hypersensitivity; GATA3 Transcription Factor; Interleukin-4; Intestinal Mucosa; Intestines; Mice; Mice, Inbred BALB C; Ovalbumin; Plants, Medicinal; T-Lymphocytes, Regulatory; Th1 Cells; Th2 Cells

It is proposed that probiotics have a therapeutic effect on the treatment of immune disorders. However, the underlying mechanisms require clarification. The present study aimed to evaluate the effect of gavage-feeding Bifidobacteria on suppression of T helper 2 (Th2) pattern inflammation in the intestines of mice with food allergy.. Mice were sensitized to ovalbumin to induce the intestinal Th2 pattern inflammation. Allergic mice were treated with or without Bifidobacteria via gavage-feeding. Th2 response, number of regulatory T cells (Treg) in the spleen and intestine, intestinal epithelial barrier function and bifidobacterial translocation were examined.. The results showed that serum-specific immunoglobulin E antibody and interleukin 4 (IL-4) were increased in allergic mice. Intestinal epithelial barrier function was impaired in allergic mice as shown by the increase in epithelial ion secretion and permeability to macromolecular protein horseradish peroxidase in Ussing chambers. Number of Treg was decreased in both spleen and intestines of allergic mice. Gavage-feeding Bifidobacteria significantly suppressed the skewed Th2 response and increased the number of Treg. Transient bifidobacterial translocation was observed in allergic mice.. Oral administration of Bifidobacteria has the capacity to suppress the skewed Th2 response in allergic mice, increasing the number of Treg and IL-10-positive cells and improve the impaired intestinal epithelial barrier function.

Topics: Administration, Oral; Animals; Bacterial Translocation; Bifidobacterium; Dendritic Cells; Disease Models, Animal; Enterotoxins; Female; Food Hypersensitivity; Green Fluorescent Proteins; Immunoglobulin E; Interleukin-10; Interleukin-4; Intestinal Mucosa; Intestines; Jejunum; Mice; Mice, Inbred BALB C; Ovalbumin; Permeability; Probiotics; T-Lymphocytes, Regulatory; Th2 Cells

To study the feasibility of C3H/HeJ mice as the food allergenicity rodent model and explore the appropriate period of two different routes of exposure and the change of helper T lymphocyte cell.. Allergenic protein ovalbumin (OVA) was administered intraperitoneally or orally to female C3H/HeJ mice for 35 days. Blood samples were taken on the 14th, 28th and 35th days from the orbital plexus and sera were obtained to determine OVA specific IgE (OVA sIgE). Spleen samples were obtained on the 35th day to detect the mRNA expression of Th2-type cytokines IL-4 and Th1-type cytokines IFN-gamma by real-time RT-PCR.. Compared with the control group, the concentration of OVA sIgE on 28th and 35th days were significantly increased in OVA group by gavage (P < 0.05). The concentration of OVA sIgE on 14th, 28th and 35th days were significantly higher than the control group when OVA was given intraperitoneally (P < 0.05). Compared with the control group, the mRNA expression of IL-4 of OVA group and OVA + Al(OH)3 group were significantly increased. The mRNA expression of IFN-gamma of OVA group and OVA + Al(OH) group were significantly lower than the control group.. C3H/HeJ mice might be used as a rodent model for the assessment of allergenicity of food. The appropriate period is the 14th day for the route of intraperitoneal injection and the 28th day for oral gavage, respectively.

Topics: Animals; Disease Models, Animal; Female; Food Hypersensitivity; Immunoglobulin E; Interferon-gamma; Interleukin-4; Mice; Mice, Inbred C3H; Ovalbumin; T-Lymphocytes, Helper-Inducer

Thymic stromal lymphopoietin (TSLP) is a cytokine produced by epithelial cells that acts on dendritic cells, mast cells, T cells, and B cells. TSLP is involved in the pathogenesis of allergic inflammation in the lung and skin, but data indicate a regulatory role in the gastrointestinal tract. We tested the functional role of TSLP in mouse models of gastrointestinal allergy and tolerance.. TSLP Receptor (TSLPR)(+/+) and TSLPR(-/-) mice were sensitized and challenged with ovalbumin; models of allergic diarrhea or systemic anaphylaxis were studied. To induce oral tolerance, mice were fed with low-dose ovalbumin before they were immunized with it. Tolerance was measured from inhibition of ear swelling in a delayed-type hypersensitivity reaction.. TSLPR(-/-) mice were protected from the onset of allergic diarrhea; they did not develop mastocytosis in the jejunum and had reduced ovalbumin-immunoglobulin E in their serum, compared with TSLPR(+/+) mice. TSLPR(-/-) mice also lost T helper cell (Th) 2-mediated inflammation in the jejunum. In contrast, sensitization and oral tolerance were not impaired in TSLPR(-/-) mice. Transfer of wild-type, Th2-primed cells to TSLPR(-/-) mice completely restored the development of allergic diarrhea. Antigen presentation assays showed that TSLPR on T cells, but not dendritic cells, was required to mediate the Th2 response.. TSLP is required for allergic inflammation but not primary sensitization or tolerance to food proteins in the gastrointestinal tract; it amplifies Th2 responses directly from CD4(+) T cells.

Topics: Anaphylaxis; Animals; Cytokines; Dendritic Cells; Diarrhea; Food Hypersensitivity; Gastrointestinal Tract; Immune Tolerance; Mice; Mice, Inbred BALB C; Ovalbumin; Th2 Cells; Thymic Stromal Lymphopoietin

Colonization of the gut by commensal bacteria modulates the induction of oral tolerance and allergy. However, how these intestinal bacteria modulate antigen-specific T cell responses induced by oral antigens remains unclear. In order to investigate this, we used germ-free (GF) ovalbumin (OVA)-specific T cell receptor transgenic (OVA23-3) mice. Conventional (CV) or GF mice were administered an OVA-containing diet. Cytokine production by CD4(+) cells from spleen (SP), mesenteric lymph nodes (MLN) and Peyer's patches (PP) was evaluated by ELISA, as was the peripheral antibody titer. T cell phenotype was assessed by flow cytometry. CD4(+) cells from the SP and MLN of CV and GF mice fed an OVA diet for 3 weeks produced significantly less IL-2 than the corresponding cells from mice receiving a control diet, suggesting that oral tolerance could be induced at the T cell level in the systemic and intestinal immune systems of both bacterial condition of mice. However, we also observed that the T cell hyporesponsiveness induced by dietary antigen was delayed in the systemic immune tissues and was weaker in the intestinal immune tissues of the GF mice. Intestinal MLN and PP CD4(+) T cells from these animals also produced lower levels of IL-10, had less activated/memory type CD45RB(low) cells, and expressed lower levels of CTLA-4 but not Foxp3 compared to their CV counterparts. Furthermore, GF mice produced higher serum levels of OVA-specific antibodies than CV animals. CD40L expression by SP CD4(+) cells from GF mice fed OVA was higher than that of CV mice. These results suggest that intestinal commensal bacteria promote T cell hyporesponsiveness and down-regulate serum antibody responses induced by dietary antigens through modulation of the intestinal and systemic T cell phenotype.

Topics: Animals; Antibodies; Antigens; Dietary Proteins; Down-Regulation; Food Hypersensitivity; Germ-Free Life; Humans; Intestines; Mice; Mice, Inbred BALB C; Mice, Transgenic; Ovalbumin; Receptors, Antigen, T-Cell; T-Lymphocytes

Red Ginseng roots (Panax ginseng C.A. Meyer) have traditionally been thought to have anti-allergic effects, but their influence on food-induced allergic responses is unclear.. This study examined the effects of a Red Ginseng extract on an ova-albumin (OVA)-evoked allergic reaction in mice.. The orally administered extract significantly inhibited the increase in OVA-specific IgG(1) (Th(2)) levels in OVA-sensitized mice, but had no effect on OVA-specific IgE (Th(2)) levels. The extract prevented a reduction in IL-12 production and the ratio of IFN-γ (Th(1)) to IL-4 (Th(2)) in splenocytes, and enhanced small intestinal CD8-, IFN-γ-, and IgA-positive cell numbers in the OVA-sensitized mice. These findings suggest that Red Ginseng inhibits allergic reactions to food by preventing reductions in the ratio of IFN-γ to IL-4 and in IL-12 production induced by dietary antigens in spleen cells, and/or increasing the expression of CD8 and IFN-γ in the small intestine. It may also protect against sensitization to antigens as an immunomodulator by increasing intestinal IgA secretion without affecting antigen-specific IgE levels. In conclusion, Red Ginseng roots may be a natural preventative of food allergies.

Topics: Animals; Anti-Allergic Agents; Disease Models, Animal; Food Hypersensitivity; Immunoglobulins; Interferon-gamma; Interleukin-12; Interleukin-4; Intestine, Small; Male; Mice; Mice, Inbred BALB C; Ovalbumin; Panax; Plant Extracts; Plant Roots; Spleen

The aim of the present study was to investigate the effect of oral probiotic bacteria administration at different times on ovalbumin-sensitized rats. Brown-Norway (BN) rats were orally sensitized with ovalbumin for 6 weeks. Probiotics were administered before the initial sensitization (prevention group) or at the end of sensitization period (treatment group). In whole-course intervention group, probiotics were administered 2 weeks before the initial sensitization until 1 week after the end of sensitization period. Ovalbumin-immunoglobulin E (IgE) level, intestinal barrier function and immune responses were analyzed. The positive control group had a significantly increased ovalbumin-IgE level (P<0.05), impaired intestinal barrier function and skewed T-helper 1 (Th1)/Th2 cytokine balance compared with the negative control group. In probiotics prevention and whole-course intervention groups, the infiltration of inflammatory cells (eosinophil and mast cells) in small intestinal mucosa was significantly lower (P<0.05), and the ratio of cytokine interferon-γ/interleukin-4 produced by spleen and mesenteric lymph nodes significantly higher (P<0.05) than in the positive control group, which suggested a cytokine profile inclined to Th1. Both probiotics prevention and prebiotics treatment could attenuate food allergic response. Probiotics prevention tends to modulate the immune response, whereas probiotics treatment has a more obvious effect in enhancing intestinal integrity.

Topics: Animals; Bacterial Load; Bifidobacterium; Colon; Cytokines; Disease Models, Animal; Eosinophils; Female; Food Hypersensitivity; Immunoglobulin A, Secretory; Immunoglobulin E; Interferon-gamma; Interleukin-4; Intestinal Mucosa; Intestine, Small; Lacticaseibacillus rhamnosus; Lymph Nodes; Mast Cells; Ovalbumin; Permeability; Probiotics; Rats; Rats, Inbred BN; Th1-Th2 Balance

To investigate the effects of fulvotomentoside (Ful) on inflammatory factors and antiinflammatory factors in intestine of ovalbumin (OVA)-sensitized BALB/c mice, and to explore the mechanisms of its anti-food allergy effect.. Twenty-four female BALB/c mice aged 6 weeks fed with ovalbumin-free feed were randomly divided into 3 groups, food allergy (FA) group, Ful group and normal saline (NS) group. Mice in FA and Ful groups were sensitized intraperitoneally two times with OVA and challenged intragastrically with OVA. Mice in Ful group were treated with 200 mg/kg of Ful by subcutaneous injection once daily for 22 days. The mice in FA and NS groups were used as positive control and negative control, respectively, and were treated with normal saline solution by subcutaneous injection for 22 days. Just 48 hours after the last challenge, the mice in each group were sacrificed and specimens of jejunum were taken. The mRNA expressions of transforming growth factor β1 (TGF-β1), interleukin-6 (IL-6), interleukin-17A (IL-17A) and forkhead box P3 (Foxp3) in jejunum were detected by reverse transcription-PCR (RT-PCR). The protein expressions of TGF-β1, IL-6, and IL-17A in jejunum were detected by immunohistochemical method. The activation of neutrophils in jejunum was assayed by the levels of MPO.. The expressions of TGF-β1, IL-6, IL-17A mRNA [(0.370 ± 0.013), (0.475 ± 0.015), (0.541 ± 0.013)] and related protein [(53,075.70 ± 20,727.06), (256,881.66 ± 36,561.79), (435,064.25 ± 69,911.48)] in jejunum were increased and the Foxp3 mRNA [(0.231 ± 0.014) vs. (0.365 ± 0.015)] expression was decreased in group FA. After the treatment with Ful, IL-6 and IL-17A mRNA [(0.196 ± 0.005), (0.204 ± 0.008)] and protein [(114,040.30 ± 20,295.25), (218,200.74 ± 30,077.69)] expressions were decreased and Foxp3 mRNA (0.578 ± 0.021) expression was increased, and no change of TGF-β1 expression was unchanged. There were no significant differences of the levels of MPO among the three groups.. Inflammatory reaction which was characterized by the increase of IL-6 and IL-17A expressions was found in intestine of ovalbumin-sensitized BALB/c mice. Ful could decrease overexpression of IL-6 and IL-17A, and increase the expression of specific transcription factor Foxp3 of regulatory T cells significantly in intestine. It may be one of the mechanisms that Ful improved intestinal inflammation.

Topics: Animals; Female; Food Hypersensitivity; Forkhead Transcription Factors; Inflammation; Interleukin-17; Interleukin-6; Intestinal Mucosa; Intestines; Mice; Mice, Inbred BALB C; Oleanolic Acid; Ovalbumin; Saponins; Transforming Growth Factor beta1

Nitration of proteins on tyrosine residues, which can occur due to polluted air under "summer smog" conditions, has been shown to increase the allergic potential of allergens. Since nitration of tyrosine residues is also observed during inflammatory responses, this modification could directly influence protein immunogenicity and might therefore contribute to food allergy induction. In the current study we have analyzed the impact of protein nitration on sensitization via the oral route.. BALB/c mice were immunized intragastrically by feeding untreated ovalbumin (OVA), sham-nitrated ovalbumin (snOVA) or nitrated ovalbumin (nOVA) with or without concomitant acid-suppression. To analyze the impact of the sensitization route, the allergens were also injected intraperitoneally. Animals being fed OVA or snOVA under acid-suppressive medication developed significantly elevated levels of IgE, and increased titers of specific IgG1 and IgG2a antibodies. Interestingly, oral immunizations of nOVA under anti-acid treatment did not result in IgG and IgE formation. In contrast, intraperitoneal immunization induced high levels of OVA specific IgE, which were significantly increased in the group that received nOVA by injection. Furthermore, nOVA triggered significantly enhanced mediator release from RBL cells passively sensitized with sera from allergic mice. Gastric digestion experiments demonstrated protein nitration to interfere with protein stability as nOVA was easily degraded, whereas OVA and snOVA remained stable up to 120 min. Additionally, HPLC-chip-MS/MS analysis showed that one tyrosine residue (Y(107)) being very efficiently nitrated is part of an ovalbumin epitope recognized exclusively after oral sensitization.. These data indicated that despite the enhanced triggering capacity in existing allergy, nitration of OVA may be associated with a reduced de novo sensitizing capability via the oral route due to enhanced protein digestibility and/or changes in antibody epitopes.

Topics: Air Pollution; Allergens; Amino Acid Sequence; Animals; Chromatography, High Pressure Liquid; Disease Models, Animal; Epitopes; Female; Food Hypersensitivity; Immunoglobulin E; Immunoglobulin G; Inflammation; Mice; Mice, Inbred BALB C; Molecular Sequence Data; Nitrogen; Ovalbumin; Smog; Tandem Mass Spectrometry; Tyrosine

The number of patients with food allergy has increased dramatically over the last several decades. However, there is no effective drug for food allergies. In the present study, we evaluated the effects of kakkonto, a traditional Japanese herbal medicine, in a mouse model of food allergy with gastrointestinal symptoms.. BALB/c mice were systemically sensitized twice with ovalbumin (OVA) and then were repeatedly given OVA by oral intubation (OVA mice). Kakkonto was administered orally before the OVA challenges.. The OVA mice developed allergic diarrhea (91.8 +/- 3.8% after 6 OVA challenges), and myeloperoxidase (MPO) activity was dramatically elevated in the colons of the OVA mice. Kakkonto significantly suppressed the occurrence of allergic diarrhea and MPO activity in the OVA mice. Furthermore, the number of mucosal mast cells was greatly increased in the proximal colons of the OVA mice, and this was also suppressed by kakkonto. Interestingly, mRNA expression of helper T cell type 1 (Th1) cytokines (IFN-gamma) and Th2 cytokines (IL-4, IL-5 and IL-10) were significantly upregulated in the proximal colons of the OVA mice, an effect which was also reduced by kakkonto. Transcriptome analysis detected increased mRNA expression of suppressor of cytokine signaling-3 in the proximal colons of OVA mice, which was decreased by kakkonto administration.. Kakkonto has immunosuppressive effects and interferes with the infiltration of mucosal mast cells in the colons of mice with induced food allergy, leading to improvement of allergic symptoms. Kakkonto has potential as a therapeutic drug for treatment of allergic symptoms induced by the disruption of intestinal mucosal immunity.

Topics: Anaphylaxis; Animals; Cell Movement; Chemokines; Chymases; Colon; Diarrhea; Disease Models, Animal; Drugs, Chinese Herbal; Food Hypersensitivity; Gene Expression; Gene Expression Profiling; Immunoglobulin E; Interleukins; Intestinal Mucosa; Male; Mast Cells; Mice; Mice, Inbred BALB C; Ovalbumin; Peroxidase; Phytotherapy; Suppressor of Cytokine Signaling 3 Protein; Suppressor of Cytokine Signaling Proteins

Mice allergic to ovalbumin (OVA) avoid drinking a solution containing this antigen. This was interpreted as related to IgE-dependent mast cell degranulation and sensory C fiber activation.. We employed pharmacological manipulation to further investigate the mediators involved in immune-induced food aversion.. While nonimmunized rats preferred a sweetened OVA solution, immunized rats avoided it. We also employed a paradigm in which rats are conditioned to drink water for two 10-min sessions a day. Tolerant rats presented lower IgE titers, and this manipulation abrogated food aversion. Dexamethasone (1.0 mg/kg) prevented the aversion of OVA-immunized rats to the antigen-containing solution. Combined blockade of H(1) and 5-hydroxytryptamine (5-HT)(2) receptors by promethazine (3.0 mg/kg) plus methysergide (5.0 mg/kg) was unable to alter food aversion. Blockade of 5-HT(3) receptors by ondansetron (1.0 mg/kg) caused a twofold increase in the ingestion of the sweetened OVA solution by immunized rats, suggesting the involvement of 5-HT(3) receptors in food aversion. Finally, we showed that dexamethasone or promethazine plus methysergide, but not ondansetron, effectively prevented the IgE-dependent mast-cell-degranulation-induced increase in vascular permeability in rats.. We suggest that regardless of whether or not they cause edema, IgE-mediated mast cell degranulation and consequent 5-HT(3) signaling are involved in the process that triggers avoidance to the source of the allergen in allergic rats.

Topics: Animals; Avoidance Learning; Capillary Permeability; Cell Degranulation; Conditioning, Psychological; Dexamethasone; Disease Models, Animal; Feeding Behavior; Food Hypersensitivity; Immunoglobulin E; Immunosuppressive Agents; Male; Mast Cells; Methysergide; Ondansetron; Ovalbumin; Rats; Rats, Wistar; Serotonin; Serotonin 5-HT3 Receptor Antagonists; Serotonin Antagonists

Food allergy can cause food-related anaphylaxis. Food allergen labeling is the principal means of protecting sensitized individuals. This motivated European Directive 2003/89 on the labeling of ingredients or additives that could trigger adverse reactions, which has been in effect since 2005. During this study, we developed animal models with allergy to ovalbumin, caseinate, and isinglass in order to be able to detect fining agent residues that could induce anaphylactic reactions in sensitized mice. The second aim of the study was to design sandwich ELISA tests specific to each fining agent in order to detect their residue antigenicity, both during wine processing and in commercially available bottled wines. Sensitized mice and sandwich ELISA methods were established to test a vast panel of wines. The results showed that although they were positive to our highly sensitive sandwich-ELISA tests, some commercially available wines are not allergenic in sensitized mice. Commercially available bottled wines made using standardized processes, fining, maturation, and filtration, do not therefore represent any risk of anaphylactic reactions in sensitized mice.

Topics: Allergens; Anaphylaxis; Animals; Caseins; Cattle; Chickens; Disease Models, Animal; Enzyme-Linked Immunosorbent Assay; Female; Fishes; Food Hypersensitivity; Gelatin; Humans; Mice; Mice, Inbred Strains; Ovalbumin; Rabbits; Wine

Studies suggested that an increased transforming growth factor-beta (TGF-beta) level in the intestinal tract helps to alleviate symptoms of food allergy. An efficient method for in vivo delivery of TGF-beta is yet to be developed. In this report, we investigated some DNA-based therapeutic strategies. Our results demonstrate that a sustainable increase of TGF-beta protein in mouse intestinal tissue can be induced after oral administration by gavage of a TGF-beta expressing DNA vector that was packed in chitosan nanoparticles. Concomitantly, a significant amelioration of ovalbumin-induced food allergy symptoms was observed. Thus, our study supports this therapy being perhaps a better alternative to the previously reported protein-based strategies.

Topics: Animals; Chitosan; Female; Food Hypersensitivity; Gene Transfer Techniques; Genetic Therapy; Histamine; Immunoglobulin A; Immunoglobulin E; Interferon-gamma; Interleukin-4; Intestinal Mucosa; Intestines; Mice; Mice, Inbred BALB C; Nanoparticles; Ovalbumin; Plasmids; Transforming Growth Factor beta1

Although CD4+ T-cell populations are thought to be involved in the pathophysiology of food allergy and oral tolerance, the role of CD8+ T cells remains uncertain.. We analyzed regulatory effects of adoptively transferred CD8+ T cells on the development of allergic diarrhea in antigen-sensitized mice that had a significantly reduced number of conventional TCRalphabeta+ CD8+ T cells.. Ovalbumin-specific T-cell receptor transgenic mice were systemically sensitized to ovalbumin. Splenic CD8+ T cells purified from ovalbumin-sensitized or nonsensitized wild-type mice or IL-10 knockout mice were adoptively transferred to ovalbumin-sensitized ovalbumin-specific T-cell receptor transgenic mice. Allergic diarrhea induced by oral administration of ovalbumin, ovalbumin-specific immunoglobulin production, and cytokine production in intestines and mesenteric lymph nodes were assessed.. Adoptive transfer of splenic CD8+ T cells from ovalbumin-primed mice, but not from nonprimed mice, suppressed the development of allergic diarrhea, which was associated with in vivo increased IL-10 mRNA expression and in vitro antigen-specific IL-10 production by mesenteric lymph node cells. Upregulation of serum ovalbumin-specific IgE was not suppressed by ovalbumin-primed CD8+ T-cell transfer. Although administration of IL-10 before ovalbumin challenge failed to alleviate allergic diarrhea, transfer of splenic CD8+ T cells from IL-10 knockout mice showed diminished preventive effects.. Systemic immunization with allergen simultaneously induces regulatory CD8+ T cells that can inhibit the development of allergic diarrhea. IL-10 production by regulatory CD8+ T cells appears to be partially involved in these inhibitory mechanisms.

Topics: Adoptive Transfer; Animals; CD8-Positive T-Lymphocytes; Cells, Cultured; Cytokines; Diarrhea; Food Hypersensitivity; Immunoglobulin E; Interleukin-10; Mice; Mice, Inbred BALB C; Mice, Transgenic; Ovalbumin; Receptors, Antigen, T-Cell; Spleen

Aluminium (ALUM) is used as experimental and clinical adjuvant for parenteral vaccine formulation. It is also contained in anti-acid drugs like sucralfate (SUC). These anti-acids have been shown to cause sensitization to food proteins via elevation of the gastric pH. The aim of this study was to assess the oral adjuvant properties of ALUM, alone or contained in SUC, in a BALB/c mouse model.. Mice were fed SUC plus ovalbumin (OVA) and compared with groups where ALUM or proton pump inhibitors (PPI) were applied as adjuvants. The humoral and cellular immune responses were assessed on antigen-specific antibody and cytokine levels. The in vivo relevance was investigated in skin tests.. The highest OVA-specific immunoglobulin G1 (IgG1) and IgE antibody levels were found in mice fed with OVA/SUC, followed by OVA/ALUM-treated animals, indicating a T helper 2 (Th2) shift in both groups. Antibody levels in other groups revealed lower (OVA/PPI-group) or baseline levels (control groups). Positive skin tests confirmed an allergic response in anti-acid or adjuvant-treated animals.. Our data show for the first time that ALUM acts as a Th2-adjuvant via the oral route. This suggests that orally applied SUC leads to an enhanced risk for food allergy, not only by inhibiting peptic digestion but also by acting as a Th2-adjuvant by its ALUM content.

Topics: Adjuvants, Immunologic; Administration, Oral; Alum Compounds; Animals; Antacids; Female; Food Hypersensitivity; Gastric Acidity Determination; Immunoglobulin E; Immunoglobulin G; Interferon-gamma; Interleukin-5; Mice; Mice, Inbred BALB C; Ovalbumin; Skin Tests; Sucralfate; Th2 Cells

To develop an oral-sensitized animal model of food allergy using Brown-Norway (BN) rats and evaluate the sensitivity of ELISA and passive cutaneous anaphylaxis (PCA) in detecting ovalbumin-specific IgE antibody (OVA-IgE) level in sensitized animals.. Sixteen 3-week old female BN rats were randomly divided into 3 groups: negative control group orally gavaged with saline, positive control group sensitized by intraperitoneal injection of 0. lmg/d OVA, and, study group sensitized by daily gavage of 1 mg/d ovalbumin (OVA). OVA-IgE was analyzed by ELISA and PCA method at week 4, 5, 6, 7, 8 and 9. At week 13, OVA-IgE level was analyzed after orally challenged by 1.0 ml of 100 mg/ml OVA.. The ELISA result showed that the OVA-IgE level in study group was significantly increased at week 6, 7 and week 8 compared with negative control group (P < 0.05), and the highest level was found at week 6. There was no significant difference for the level of OVA-IgE between study group and positive control group. The sensitization rate in study group was 60%, 80% and 80% at week 6, 7 and 8 respectively, which was similar to positive control group. All PCA results in study group were negative, while in positive control group it was positive.. Oral sensitization could be used as a suitable method to establish an animal model of food allergy, which is more comparable with the natural sensitization process in food allergy patients. ELISA method is more sensitive in detecting OVA-IgE level in oral sensitized animal model than PCA method.

Topics: Administration, Oral; Allergens; Animals; Disease Models, Animal; Female; Food Hypersensitivity; Immunoglobulin E; Ovalbumin; Random Allocation; Rats; Rats, Inbred BN

Cancer is the leading cause of mortality worldwide. Analysis of epidemiological data has revealed a negative relationship between allergic conditions and cancer incidence. This study addresses the effects of chronic antigen ingestion by sensitized mice (allergy) on Ehrlich tumor growth in mouse footpad. Mice were sensitized (allergic) or not (sham) with ovalbumin and challenged orally with egg white solution. After one week of oral challenge, all mice were inoculated with experimental Ehrlich tumor (EET) cells in the footpad, and tumor growth was evaluated for 21 days. A decrease in tumor growth occurred, as assessed by paw thickness in the allergic group, which was associated with smaller areas of necrosis, reduced infiltration of neutrophils, and reduced levels of IFN-gamma, IL-4, and IL-10. Although, the tumor proliferation rate was similar in both groups, an increase in apoptosis occurred in allergic mice. In conclusion, analysis of the data obtained allows us to suggest that a concomitant allergic condition would reduce tumor progression through increased tumor cell apoptosis, accompanied by reduced areas of necrosis at the tumor site. Indeed, such findings suggested a possible mechanism for the reduced cancer incidence observed in allergic individuals.

Topics: Animals; Apoptosis; Carcinoma, Ehrlich Tumor; Cell Count; Cytokines; Disease Models, Animal; Enzyme-Linked Immunosorbent Assay; Female; Food Hypersensitivity; Foot; Immunoglobulin E; Immunoglobulin G; In Situ Hybridization; Mice; Mice, Inbred BALB C; Neutrophils; Ovalbumin; Peroxidase; Xenograft Model Antitumor Assays

Mechanisms involved in the induction of oral tolerance (OT) or systemic immunization through the oral rout are still poorly understood. In our previous studies, we have shown that when normal mice eat peanuts they become tolerant, with no gut alterations. Conversely, if immunized with peanut proteins prior to a challenge diet (CD) containing peanuts they develop chronic inflammation of the gut. Our aim is to evaluate the consequences of the introduction of a novel protein in the diet of animals presenting antigen-specific gut inflammation. Adult, female C57BL/6J mice were divided in control (C) and experimental (E) groups. C1-C3 received peanut protein immunization, animals of the control groups C4 were sham immunized, and control group C5 received ovalbumin (OVA) immunization. The experimental group was immunized with peanut protein extract. Before initial exposure to a 30-day peanut containing CD, the experimental group was divided into 5 groups (E1-E5). OVA feeding began 7 days prior CD (E1) on day 0 (E2), 7 (E3), 14 (E4) and 21 (E5) during CD. Our results show that oral exposure to a novel protein (OVA) in the absence of gut inflammation (E1) leads to low levels of systemic antibody (Ab) titers, comparable to tolerant animals. Conversely, as off initial induction of inflammation, groups submitted to OVA (OT) protocol develop increasingly higher systemic Ab titers similar to animals of the immune control group. In conclusion, our protocol indicates that timing is more important than the antigenicity when a novel protein is offered, in the diet.

Topics: Administration, Oral; Animals; Antibody Formation; Antigens, Plant; Arachis; Diet; Epitopes; Female; Food Hypersensitivity; Humans; Immune Tolerance; Immunity, Mucosal; Immunization; Intestines; Mice; Mice, Inbred C57BL; Ovalbumin; Time Factors

Genetic engineering of food crops has significantly influenced the agricultural productivity over the past two decades. It has proved a valuable tool, offering crops with higher yields, improved nutritional quality, resistance against pesticides, herbicides and tolerance against abiotic stresses. However, the safety assessment of genetically engineered (GE) crops is prerequisite before introduction into human food chain. The present study was aimed to assess the toxicity and allergenicity of leaf curl virus resistant GE tomato compared to its wild-type species. Balb/c mice fed with genetically engineered or wild-type tomato did not show significant differences in growth, body weight (P > 0.05) and food consumption when compared with control mice. Values for serum glutamic oxaloacetic transaminase and serum glutamic pyruvic transaminase, urea and cholesterol were comparable in GE and wild-type tomato fed mice. Mice immunized with GE or wild-type tomato extract showed low IgE response. Lung histology of ovalbumin fed mice showed bronchoconstriction with eosinophilic infiltration whereas GE or wild-type tomato showed no cellular infiltration with normal airways. Genetically engineered and wild-type tomato sensitized mice demonstrated similar IL-4 release in splenic cell culture supernatant. GE and wild tomato extract on ELISA showed comparable IgE binding (P > 0.05) with food allergic patients' sera. In conclusion, genetically engineered tomato showed no toxicity in mice and allergenicity is similar to the wild-type tomato.

Topics: Allergens; Animals; Begomovirus; Food Hypersensitivity; Humans; Immunoglobulin E; Mice; Mice, Inbred BALB C; Ovalbumin; Plants, Genetically Modified; Serum; Solanum lycopersicum; Spleen

Allergic reactions to food can involve diarrhea, vomiting, nausea and abnormal pain. PG102 has previously been shown to control various factors involved in allergy pathogenesis, including IgE and various Th1 and Th2 cytokines, in vivo as well as in vitro [Park EJ, et al.: J Allergy Clin Immunol 2005;116:1151-1157; Park EJ, et al.: J Invest Dermatol 2007;127:1154-1160]. These data indicate that PG102 might have antiallergic effects on allergic diarrhea. Here, we investigated whether PG102 could prevent allergic diarrhea in the murine ovalbumin (OVA)-induced allergic diarrhea model.. BALB/c mice were orally treated with PG102, dexamethasone or water for 9 days on a daily basis, followed by subcutaneous injection with OVA on day 0. Animals were orally administrated with OVA from day 7, 3 times a week, over a period of approximately 20 days. Incidence of diarrhea, serum, OVA-restimulated splenocytes and lamina propria lymphocytes were analyzed.. Oral administration of PG102 could suppress the incidence of diarrhea in a murine allergic diarrhea model. The amelioration of allergic diarrhea by PG102 was accompanied with the inhibition of mast cell infiltration into the large intestine. The serum level of IgE, IL-6 and MCP-1 was decreased in PG102-treated mice. When splenocytes were isolated from respective groups and cultured in the presence of OVA, cells from PG102-administrated animals produced lesser amounts of IL-6 and MCP-1.. PG102 has the potential to be used as a preventive for food allergic diseases.

Topics: Actinidia; Animals; Calcimycin; Cell Movement; Chemokine CCL2; Dexamethasone; Diarrhea; Disease Models, Animal; Female; Food Hypersensitivity; Immunoglobulin E; Interleukin-10; Interleukin-4; Interleukin-6; Intestine, Large; Lymphocytes; Mast Cells; Mice; Mice, Inbred BALB C; Mucous Membrane; Ovalbumin; Plant Extracts; Rats; Spleen

We hypothesized that food allergy causes a state of non-specific jejunal dysmotility. This was tested in a mouse model.. Balb/c mice were epicutaneously sensitized with ovalbumin and challenged with 10 intragastric ovalbumin administrations every second day. Smooth muscle contractility of isolated circular jejunal sections was studied in organ bath with increasing concentrations of carbamylcholine chloride (carbachol). Smooth muscle layer thickness and mast cell protease-1 (MMCP-1) positive cell density were assayed histologically. Serum MMCP-1 and immunoglobulins were quantified by ELISA, and mRNA expressions of IFN-gamma, IL-4, IL-6 and TGFbeta-1 from jejunal and ileal tissue segments were analyzed with quantitative real-time PCR.. Ovalbumin-specific serum IgE correlated with jejunal MMCP-1+ cell density. In the allergic mice, higher concentrations of carbachol were required to reach submaximal muscular stimulation, particularly in preparations derived from mice with diarrhoea. Decreased sensitivity to carbachol was associated with increased expression of IL-4 and IL-6 mRNA in jejunum. Smooth muscle layer thickness, as well as mRNA of IFN-gamma and TGF-beta1 remained unchanged.. In this mouse model of food allergy, we demonstrated a decreased response to a muscarinic agonist, and increased levels of proinflammatory IL-6 and Th2-related IL-4, but not Th1-related IFN-gamma mRNAs in jejunum. IgE levels in serum correlated with the number of jejunal MMCP-1+ cells, and predicted diarrhoea. Overall, these changes may reflect a protective mechanism of the gut in food allergy.

Topics: Animals; Carbachol; Cholinergic Agents; Cytokines; Disease Models, Animal; Female; Food Hypersensitivity; Gastrointestinal Motility; Ileum; Inflammation; Interferon-gamma; Interleukin-4; Interleukin-6; Jejunum; Mice; Mice, Inbred BALB C; Muscle Contraction; Muscle, Smooth; Ovalbumin; Transforming Growth Factor beta1

There are presented the results of genotoxicologic, immunologic and allergologic examinations which were conducted within the framework of integrated medical and biological assessment of genetically modified rootworm Diabrotica spp.-protected maize event MIR604. Analysis of damages of DNA and structural chromosome aberrations, assessment of the allergenic potential and immunoreactive properties has not confirmed any genotoxic, allergenic and immunotoxic effect of maize event MIR604.

Topics: Anaphylaxis; Animals; Bone Marrow Cells; Chromosome Aberrations; Colon; Comet Assay; DNA Damage; Food Analysis; Food Hypersensitivity; Food, Genetically Modified; Hypersensitivity, Delayed; Liver; Male; Mice; Mice, Inbred C57BL; Mice, Inbred CBA; Ovalbumin; Plants, Genetically Modified; Rats; Rats, Wistar; Species Specificity; Toxicity Tests; Zea mays

We have developed a rolling circle amplification-procedure as a method to enhance signals of immunoassay (immuno-RCA) to detect one of the major food allergens, ovalbumin (OVA). Immuno-RCA in combination with a fluorescent dye and a circular DNA probe prepared by intramolecular ligation easily allowed the real-time detection of OVA, and could apparently detect signals from OVA with concentrations ranging from 10(-12) to 10(-7) g/mL. Therefore, sensitive and easily handled aspects of this method would contribute to an effective signal enhancement in immunoassay for food allergen detections.

Topics: Animals; Base Sequence; DNA Probes; DNA, Circular; Fluorescent Dyes; Food Hypersensitivity; Immunoassay; Nucleic Acid Amplification Techniques; Oligodeoxyribonucleotides; Ovalbumin; Signal Processing, Computer-Assisted; Time Factors

Specific allergen immunotherapy is frequently associated with adverse reactions. Several strategies are being developed to reduce the allergenicity while maintaining the therapeutic benefits. Peptide immunotherapy is one such approach. Methods for the simple and rapid identification of immunogenic epitopes of allergens (i.e. allergenic epitopes) are ongoing and could potentially lead to peptide-based vaccines. An epitope extraction technique, based on biofunctionalized magnetic microspheres self-organized under a magnetic field in a channel of a simple microfluidic device fabricated from polydimethylsiloxane, was applied in the isolation and identification of prospective allergenic epitopes. Similarly to chromatographic column separations, the easily replaceable plug of self-organized beads in the channel benefits especially from an even larger surface-to-volume ratio and an enhanced interaction of the surfaces with passing samples. Ovalbumin, the major protein of egg white and a typical representative of food allergens, was selected as the model molecule. Highly resistant ovalbumin was at first efficiently digested by a magnetic proteolytic reactor with trypsin treated with l-1-tosylamido-2-phenylethyl chloromethyl ketone and the second step, i.e. capture of allergenic epitopes from the mixture of peptides, was performed by a magnetic immunoaffinity carrier with orientedly immobilized rabbit anti-ovalbumin IgG molecules. Captured peptides were released with 0.05% trifluoroacetic acid. The elution fractions were analyzed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. The peptide fragment of ovalbumin HIATNAVLFFGR (m/z: 1345.75, position: 371-382) was identified as a relevant allergenic epitope in this way. Such a microfluidic magnetic force-based epitope extraction technique applied in the epitope mapping of ovalbumin has the potential to be a significant step towards developing safe and cost-effective epitope-based vaccines.

Topics: Allergens; Epitope Mapping; Epitopes; Food Hypersensitivity; Immunomagnetic Separation; Mass Spectrometry; Microfluidic Analytical Techniques; Microspheres; Ovalbumin; Vaccines

Recent study has demonstrated an increasing prevalence of food allergy in Korean children. Specific probiotic bacteria may promote potentially anti-allergenic processes through induction of Th1-type immunity and enhance the regulatory lymphocyte. This study investigated whether orally administrated probiotics could suppress allergic responses in an ovalbumin (OVA)-induced allergy mouse model. Thus, female C3H/HeJ mice were orally sensitized with OVA and cholera toxin for 4 weeks. Lactobacillus acidophilus AD031, Bifidobacterium lactis AD011, and L. acidophilus AD031 plus B. lactis AD011 were fed to mice from 2 weeks before the sensitization. The OVA-induced mice that were not treated with probiotics had significantly increased serum levels of OVA-specific IgE and IgG1, and OVA-specific IgA in feces. However, the mice treated with probiotics suppressed production of the OVA-specific IgE, IgG1, and IgA. The level of IL-4 was significantly lower, and the levels of INF-gamma and IL-10 were significantly higher in the mice treated with probiotics than that in the nontreated mice. The groups treated with probiotics had decreased levels of degranulated mast cells, eosinophil granules, and tail scabs. These results indicate that L. acidophilus AD031 and B. lactis AD011 might be useful for the prevention of allergy.

Topics: Animals; Bifidobacterium; Disease Models, Animal; Eosinophils; Female; Food Hypersensitivity; Histocytochemistry; Immunoglobulins; Interferon-gamma; Interleukin-10; Interleukin-4; Lactobacillus acidophilus; Mast Cells; Mice; Mice, Inbred C3H; Ovalbumin; Probiotics; Spleen

Animal models are essential for analyzing the allergenic potential of food proteins and for investigating mechanisms underlying food allergy. Based on previous studies revealing acid-suppression medication as risk factor for food allergy induction, we aimed to establish a mouse model mimicking the natural route of sensitization in patients.. The effect of acid-suppressing medication on murine gastric pH was assessed by intragastric pH measurements after two injections of a proton pump inhibitor (PPI). To investigate dose-dependency, mice were fed different concentrations of ovalbumin (OVA; 0.2, 0.5, 1.0, 2.5 or 5.0mg) either with or without anti-ulcer medication. Additionally, different routes of exposure (i.p. vs. oral) were compared in a second immunization experiment. Sera were screened for OVA-specific antibody titers (IgG1, IgG2a and IgE) in ELISA and RBL assay. Clinical reactivity was evaluated by measuring rectal temperature after oral challenge and by type I skin tests.. Two intravenous injections of PPI significantly elevated the gastric pH from 2.97 to 5.3. Only oral immunization with 0.2mg OVA under anti-acid medication rendered elevated IgG1, IgG2a and IgE titers compared to all other concentrations. Protein feeding alone altered antibody titers only marginally. Even though also i.p. immunizations induced high levels of specific IgE, only oral immunizations under anti-acids induced anaphylactic reactions evidenced by a significant decrease of body temperature.. Only low-dosage ovalbumin feedings under anti-acid medication resulted in IgE mediated food allergy. Based on this knowledge we have established a suitable food allergy model for further investigations of food adverse reactions.

Topics: Administration, Oral; Animals; Disease Models, Animal; Female; Food Hypersensitivity; Gastric Acidity Determination; Hydrogen-Ion Concentration; Immunoglobulin E; Immunoglobulin G; Interferon-gamma; Interleukin-5; Mice; Mice, Inbred BALB C; Ovalbumin; Proton Pump Inhibitors; Skin Tests

Egg proteins are responsible for one of the most common forms of food allergy, especially in children, and one of the major allergens is ovalbumin (OVA). With the aim to examine the potential of high pressure to enhance the enzymatic hydrolysis of OVA and modify its immunoreactivity, the protein was proteolyzed with pepsin under high-pressure conditions (400 MPa). Characterization of the hydrolysates and peptide identification was performed by reversed-phase high-performance liquid chromatography-tandem mass spectrometry (RP-HPLC-MS/MS). The antigenicity (binding to IgG) and binding to IgE, using the sera of patients with specific IgE to OVA, were also assessed. The results showed that, upon treatment with pepsin at 400 MPa, all of the intact protein was removed in minutes, leading to the production of hydrolysates with lower antigenicity than those produced in hours at atmospheric pressure. However, the exposure of new target residues only partially facilitated the removal of allergenic epitopes, because the hydrolysates retained residual IgG- and IgE-binding properties as a result of the accumulation of large and hydrophobic peptides during the initial stages of hydrolysis. These peptides disappeared at later stages of proteolysis, although reactivity toward IgG and IgE was not completely abolished. Some fragments identified in the hydrolysates (such as Leu124-Phe134, Ile178-Ala187, Leu242-Leu252, Gly251-Ile259, Lys322-Gly343, Phe358-Phe366, and Phe378-Pro385) carried previously identified IgE-binding epitopes. Because some of the peptides found, such as Phe358-Phe366, probably contain only one binding site for IgE, the possibility to use high pressure to tailor hydrolysates that contain mostly peptides with only one IgE-binding site, which may help the immune system to tolerate egg proteins, is suggested.

Topics: Animals; Child; Chromatography, High Pressure Liquid; Female; Food Hypersensitivity; Humans; Hydrolysis; Immunoglobulin E; Immunoglobulin G; Male; Ovalbumin; Peptide Fragments; Pressure; Protein Binding; Tandem Mass Spectrometry

Many types of fermented food are consumed in Japan. Although some are produced by plant-origin lactic-acid bacteria (LAB) fermentation, the physiological functions of such bacteria remain unclear. We therefore isolated LAB of plant origin from Kyoto pickles and determined the immunological activity of heat-killed preparations of plant-origin LAB.. The Lactobacillus pentosus strain S-PT84 was selected from among 16 LAB of plant origin as the strongest interleukin (IL)-12-inducing strain. IL-12- and IL-10-inducing activities were determined with macrophages from BALB/c mice. The in vivo immunomodulating effect of S-PT84was determined with BALB/c mice fed S-PT84. The antiallergic activity of S-PT84 was examined in ovalbumin (OVA)/alum-administered BALB/c mice.. The L. pentosus strain S-PT84 induced production of both IL-12 and IL-10 in vitro. S-PT84 enhanced splenic natural-killer activity and modulated the T helper (Th) type 1/type 2 balance toward a Th1-dominant state. In the OVA-induced allergy model, orally administered S-PT84 lowered serum IgE levels and suppressed active cutaneous anaphylaxis reaction and splenic IL-4 production. IL-10 production from splenocytes of OVA-immunized mice was upregulated by feeding S-PT84.. Despite heat-killing, S-PT84 exhibited antiallergic effects by modulating the Th1/Th2 balance and inducing regulatory T cells. The L. pentosus strain S-PT84, which is of plant origin and isolated from a traditional Japanese food, is expected to be useful for treatment of many immune diseases including allergies, tumors, infectious diseases and auto-immune diseases.

Topics: Allergens; Anaphylaxis; Animals; Anti-Allergic Agents; Cells, Cultured; Dermatitis, Atopic; Disease Models, Animal; Food Hypersensitivity; Food Microbiology; Hot Temperature; Immunoglobulin E; Immunosuppressive Agents; Interleukin-10; Interleukin-12; Killer Cells, Natural; Lactobacillus; Male; Mice; Mice, Inbred BALB C; Ovalbumin; Probiotics; Spleen; T-Lymphocytes, Regulatory; Th1 Cells; Th2 Cells; Up-Regulation

Intestinal allergic diseases are initiated by aberrant Th2-type immune responses, including elevation of IgE antibodies (Abs) and infiltration of eosinophils. However, little is known about the role of Peyer's patches (PP) in the control of allergic diseases. Using a mouse model for food allergy, we here show that mice lacking PP are more susceptible to disease development and show higher levels of antigen-specific IgE Abs than do PP-intact mice. In our study, we noted that high numbers of eosinophils infiltrated into the small intestine of PP-null mice. In contrast, the PP of intact mice contained regulatory CD4+CD25+ Foxp3+ T cells (Treg) that are known to produce high levels of IL-10, and inhibited the development of allergic diarrhea. PP-intact mice thus developed allergic diarrhea when treated with anti-CD25 or anti-IL-10 monoclonal antibody (mAb) in vivo. These studies demonstrate that PP, as the site where IL-10-producing Treg cells are created, mediate the mucosal regulatory network for the control of undesired allergic responses in the intestine.

Topics: Adoptive Transfer; Animals; Antibodies, Monoclonal; Antibody-Producing Cells; Diarrhea; Eosinophils; Food Hypersensitivity; Gene Expression; Immunization; Immunoglobulin A; Immunoglobulin E; Immunoglobulin G; Interleukin-10; Interleukin-2 Receptor alpha Subunit; Intestine, Small; Intestines; Leukocytes, Mononuclear; Mice; Mice, Inbred BALB C; Mucous Membrane; Ovalbumin; Peyer's Patches; Receptors, Interleukin-7; Reverse Transcriptase Polymerase Chain Reaction; T-Lymphocytes, Regulatory

Fructooligosaccharides (FOS) in prebiotic foods can alter intestinal immune responses. The combination of probiotics with oligosaccharides has been reported to alter intestinal flora and suggested to be beneficial against food allergy in humans.. All male Nc/jic mice used in this 8-week study were 6 weeks of age and were allotted to the following three groups: (1) the nonsensitization group; (2) the ovalbumin (OVA) sensitization +5% fructose-containing control food administration group; and (3) the OVA sensitization +5% FOS-containing food administration group. Duodenal tissues were collected and then immunohistochemically stained with monoclonal antibodies to CCR4 and CCR5. The number of mast cells and the villus edema formation rate in the duodenum were determined by image analysis.. The number of CCR4-positive cells increased significantly in Group 2 as compared with Group 1 and tended to decrease in Group 3 as compared with Group 2. Relatively few CCR5-positive cells were observed in the duodenum. FOS tended to reduce the number of CCR4-positive cells but significantly reduced the number of mast cells and the edema formation rate in the duodenum.. This study demonstrated a correlation between the number of CCR4-positive cells and villus edema formation rate. Therefore, FOS, which we inferred to show antiallergic activity for food allergy in this study and which has already been established to be safe for use as food in humans, can be considered to be potentially useful for the prevention of food allergy in pediatric patients with allergy.

Topics: Animals; Anti-Allergic Agents; Cell Count; Disease Models, Animal; Duodenal Diseases; Duodenum; Edema; Food Hypersensitivity; Immunoglobulin E; Male; Mast Cells; Mice; Microvilli; Oligosaccharides; Ovalbumin; Probiotics; Receptors, CCR4; Receptors, CCR5; Receptors, Chemokine; T-Lymphocytes, Helper-Inducer

There have been a number of reports showing that the crude beta-glucan fraction prepared from various kinds of Basidiomycetes mushrooms acts as anti-cancer and anti-allergic reagent through stimulation of IFN-gamma production. It has been reported, however, that the exposure of the airway to beta-(1,3) d-glucan, contained in house dust, indoor moulds and some bacteria, potentiates the airway allergic response. It seems likely that the discrepant effects on immune function may be related to such factors as differences of the administration route, average molecular weight and water solubility. We isolated a new low-molecular-weight (about 100 kDa) beta-glucan from Aureobasidium pullulans 1A1 strain of black yeast, and found that it had low viscosity and was water-soluble. In this study, we examined the effects of water-soluble low-molecular-weight beta-(1-->3) and 50-80% branched beta-(1-->6) glucan (LMW-beta-Glucan) isolated from A. pullulans on the ova-albumin (OVA)-treated allergic reaction in mice. Feeding standard laboratory diets containing 0.5 and 1% LMW-beta-Glucan significantly inhibited the OVA-specific IgE elevation compared to that in OVA-sensitized mice fed standard laboratory diet alone (control). Furthermore, feeding standard laboratory diets containing 0.5 and 1% LMW-beta-Glucan inhibited the reduction of IL-12 and IFN-gamma production from splenocytes and the reduction of CD8- and IFN-gamma-positive cell number in the small intestine of the OVA-sensitized mice. These findings suggest that anti-food allergic action of LMW-beta-Glucan may be due to the inducing IFN-gamma production in the small intestine and splenocytes.

Topics: Animal Feed; Animals; Anti-Allergic Agents; Antineoplastic Agents; Basidiomycota; Carbohydrate Sequence; Cytokines; Dose-Response Relationship, Drug; Down-Regulation; Food Hypersensitivity; Glucans; Hypersensitivity; Immunoglobulin E; Immunologic Factors; Male; Mice; Mice, Inbred BALB C; Molecular Weight; Ovalbumin; Solubility; Spleen; Water

Bisphenol A (BPA) is a representative endocrine disruptor that may have adverse effects on human health. Since the development of oral tolerance during infancy may play an important role in the prevention of food allergies, we examined whether transmaternal exposure to BPA influences the development of oral tolerance. To measure antigen-specific responses, female wild-type mice mated with male ovalbumin (OVA)-specific T-cell receptor transgenic (TCR-tg) mice were fed with BPA during pregnancy and while nursing. OVA was administered to OVA-TCR-tg offspring during their weaning period. Oral administration of both high and low doses of OVA suppressed OVA-specific cell proliferation and cytokine production in both BPA-exposed and nonexposed control mice, but the OVA-mediated suppression was significantly more diminished by the BPA exposure. The accumulation of CD4+CD25+Foxp3+ T cells was diminished in the BPA-exposed offspring. Moreover, after low dose OVA administration, serum OVA-specific IgG1 and IgG2a levels were higher in the BPA-exposed offspring than in nonexposed ones. Taken together, our results indicate that transmaternal exposure to BPA seems to modulate the mechanisms underlying tolerance induction; therefore, BPA may partially interrupt the development of oral tolerance.

Topics: Animals; Benzhydryl Compounds; Clonal Deletion; Endocrine Disruptors; Estrogens, Non-Steroidal; Female; Fetus; Food Hypersensitivity; Humans; Immune Tolerance; Immunoglobulin G; Male; Maternal Exposure; Mice; Ovalbumin; Phenols; Pregnancy; Random Allocation; Spleen; T-Lymphocytes

Food allergies have become increasingly prevalent during the past few decades. Diarrhea is one of the most frequent intestinal symptoms caused by food allergens and is characterized by imbalanced ion exchange and water transfer; however, the underlying mechanism of allergic diarrhea remains unclear. Water transfer across the intestinal epithelial membrane seems to occur via aquaporins (AQPs). However, the molecular mechanism of water transfer and the pathophysiological roles of aquaporins in the intestine have not been fully established. The present studies have focused on the alterations of AQPs in a mouse model of allergic diarrhea in which BALB/c mice developed diarrhea following repeated challenges of orally administered ovalbumin. Quantitative real-time PCR analysis and immunohistochemical technique were used for expression of mRNA and protein of AQPs, respectively. AQP4 and AQP8 mRNA levels were significantly decreased in the proximal colon of allergic mice compared to controls; likewise, expression of AQP4 and AQP8 proteins was reduced in the proximal colon of the allergic mice. These results suggest that allergic diarrhea is associated with a downregulation in AQP4 and AQP8 expression.

Topics: Anaphylaxis; Animals; Aquaporin 4; Aquaporins; Colon; Diarrhea; Down-Regulation; Food Hypersensitivity; Immunohistochemistry; Injections, Intraperitoneal; Male; Mice; Mice, Inbred BALB C; Ovalbumin; RNA, Messenger

Topics: Animals; CD4-Positive T-Lymphocytes; Food Hypersensitivity; Gastrointestinal Tract; Humans; Mice; Ovalbumin; T-Lymphocytes, Regulatory

A subset of food-allergic patients does not only respond clinically with symptoms in the gastro-intestinal tract but also with asthmatic reactions.. The aim of this study was to analyse whether CD4+ T cells from mice with intestinal immediate-hypersensitivity reactions to food allergen are involved in the development of experimental asthma.. BALB/c mice were intraperitoneally sensitized to ovalbumin (OVA), followed by repeated intra-gastric (i.g.) OVA challenges. Control animals were either sham-sensitized or sham-challenged with phosphate-buffered saline (PBS). Duodenum, jejunum, ileum and colon were histologically examined. CD4+ T cells from mesenteric lymph nodes were transferred from various donor groups into recipient mice that received either OVA or PBS aerosol challenges. Recipients were analysed by measurements of lung function using head-out body-plethysmography and examination of broncho-alveolar lavage and lung histology.. The highest levels of OVA-specific IgE antibody levels were detected in OVA-sensitized and OVA-challenged mice. Throughout the lower intestinal tract, a marked infiltration with eosinophils was observed, and goblet cell numbers as well as goblet cell area were significantly increased. The villus/crypt ratio was decreased compared with controls. The transfer of CD4+ T cells from mesenteric lymph nodes of OVA-sensitized and OVA-challenged mice triggered airway hyperreactivity and eosinophilic airway inflammation in recipients aerosol challenged with OVA, but not with PBS.. We conclude that CD4+ T cells from mesenteric lymph nodes of mice with allergen-induced immediate-type hypersensitivity reactions in the gut are able to transfer the phenotype of experimental asthma.

Topics: Animals; Asthma; Bronchial Hyperreactivity; CD4-Positive T-Lymphocytes; Female; Food Hypersensitivity; Hypersensitivity, Immediate; Immunoglobulin E; Intestines; Lymph Nodes; Mice; Mice, Inbred BALB C; Ovalbumin

Food allergy accounts for significant morbidity. The etiology and immune mechanisms of food allergy, however, have remained poorly understood. In this study, we aimed to determine the role of T-cell immunoglobulin-domain and mucin-domain (TIM)-4, a recently identified member of cell surface molecules, in the pathogenesis of intestinal allergy in a murine model.. We report that TIM-4 as well as costimulatory molecules were up-regulated in intestinal mucosal dendritic cells by in vitro or in vivo exposure to Staphylococcus enterotoxin B (SEB). SEB-conditioned intestinal dendritic cells loaded with a food macromolecule ovalbumin (OVA) induced potent OVA-specific T-helper (Th)2 lymphocyte responses in vitro and such Th2 responses were inhibited completely by TIM-4 blockade.. In vivo exposure to both SEB and OVA resulted in OVA-specific Th2 differentiation and intestinal allergic responses including increased serum immunoglobulin E and Th2 cytokine levels, activation of OVA-specific Th2 cells detected both ex vivo and in situ, and mast cell degranulation. Of importance, in vivo abrogation of TIM-4 or its cognate ligand TIM-1 by using a polyclonal antibody remarkably dampened Th2 differentiation and intestinal allergy.. Our study thus identifies TIM-4 as a novel molecule critically required for the development of intestinal allergy.

Topics: Animals; Cell Differentiation; Cells, Cultured; Dendritic Cells; Disease Models, Animal; Enterotoxins; Food Hypersensitivity; Hepatitis A Virus Cellular Receptor 1; Immunoglobulin E; Intestinal Mucosa; Male; Membrane Proteins; Mice; Mice, Inbred BALB C; Mice, Inbred C57BL; Mice, Transgenic; Ovalbumin; Th2 Cells

Topics: Animals; Cell Differentiation; Dendritic Cells; Disease Models, Animal; Enterotoxins; Food Hypersensitivity; Hepatitis A Virus Cellular Receptor 1; Immunoglobulin E; Intestinal Mucosa; Membrane Proteins; Mice; Ovalbumin; Th2 Cells

For monitoring the occurrence of IgE antibody specific for novel proteins in genetically modified (GM) foods, ELISA is the most convenient method. The levels of IgE specific for recombinant proteins, phosphinothricin-N-acetyltransferase (PAT), CP4-EPSPS, and Cry9C were determined by ELISA using the sera from patients allergic to known allergens. Ovalbumin (OVA) and OVA-positive patient sera were used as positive control. In the ELISA, 20-fold-diluted sera tested were mostly negative for the specific IgE. However, the PAT-specific, but not CP4-EPSPS- or Cry9C-specific IgE in some patients was apparently higher than that of the healthy volunteers. To clarify the binding specificity of the antibody, we pre-incubated the sera with soluble PAT, but the inhibition was marginal, suggesting that the binding was non-specific. Therefore, we used 1M NaCl as a washing buffer to remove IgE non-specifically bound to the coated PAT. This washing step efficiently decreased non-specific binding. In contrast, OVA-specific IgE binding to OVA-coated plate was not affected by the washing. Finally, in this pilot study significant levels of IgE antibodies specific for the three proteins were not detected in the sera of Japanese food-allergy patients.

Topics: 3-Phosphoshikimate 1-Carboxyvinyltransferase; Acetyltransferases; Allergens; Bacillus thuringiensis Toxins; Bacterial Proteins; Bacterial Toxins; Endotoxins; Environmental Exposure; Enzyme-Linked Immunosorbent Assay; Food Hypersensitivity; Glycine max; Hemolysin Proteins; Humans; Immunoglobulin E; Ovalbumin; Pest Control, Biological; Plants, Genetically Modified; Recombinant Proteins

Controversy still exists as to whether gastrointestinal colonisation by Candida albicans contributes to aggravation of atopic dermatitis. We hypothesised that Candida colonisation promotes food allergy, which is known to contribute to a pathogenic response in atopic dermatitis. We tested this using a recently established murine Candida colonisation model.. Candida colonisation in the gastrointestinal tract was established by intragastric inoculation with C albicans in mice fed a synthetic diet. To investigate sensitisation against food antigen, mice were intragastrically administered with ovalbumin every other day for nine weeks, and antiovalbumin antibody titres were measured weekly. To examine gastrointestinal permeation of food antigen, plasma concentrations of ovalbumin were measured following intragastric administration of ovalbumin.. Ovalbumin specific IgG and IgE titres were higher in BALB/c mice with Candida colonisation than in normal mice. Gastrointestinal permeation of ovalbumin was enhanced by colonisation in BALB/c mice. Histological examination showed that colonisation promoted infiltration and degranulation of mast cells. Candida colonisation did not enhance ovalbumin permeation in mast cell deficient W/Wv mice but did in congenic littermate control +/+ mice. Reconstitution of mast cells in W/Wv mice by transplantation of bone marrow derived mast cells restored the ability to increase ovalbumin permeation in response to Candida colonisation.. These results suggest that gastrointestinal Candida colonisation promotes sensitisation against food antigens, at least partly due to mast cell mediated hyperpermeability in the gastrointestinal mucosa of mice.

Topics: Animals; Antibodies; Candida albicans; Candidiasis; Enzyme-Linked Immunosorbent Assay; Female; Food Hypersensitivity; Gastric Mucosa; Gastrointestinal Diseases; Immunoglobulin A; Immunoglobulin G; Intestinal Absorption; Intestinal Mucosa; Mast Cells; Mice; Mice, Inbred BALB C; Mice, Mutant Strains; Ovalbumin; Specific Pathogen-Free Organisms

Assessing the allergenicity and toxicity of genetically modified (GM) crops is essential before they become a regular part of our food supply. The present study aimed to assess the allergenicity of Brassica juncea (mustard) expressing choline oxidase (codA) gene from Arthrobacter globiformis that provides resistance against abiotic stresses.. SDAP, Farrp, and Swiss-Prot databases were used to study allergenicity of choline oxidase. Digestibility of choline oxidase was assessed in simulated gastric fluid (SGF). Specific immunoglobulin E (IgE) reactivity of native and GM mustard was compared by using enzyme-linked immunosorbent assay (ELISA) and skin tests in respiratory-allergic patients. Allergenicity of GM and native mustard proteins was compared in Balb/c mice.. Choline oxidase showed no significant homology with allergenic proteins in SDAP and Farrp databases. Cross-reactive epitope search showed a stretch similar to Hev b 6 having some antigenic properties. Purified choline oxidase showed complete degradation with SGF. Skin prick test of native and GM mustard extract on respiratory allergic patients showed significant correlation (P < 0.05). ELISA with 96 patients' sera showed comparable IgE reactivity. Balb/c mice immunized with native and GM mustard proteins showed low IgE response. Presensitized mice on intravenous challenge with Brassica extract showed no anaphylactic symptoms unlike ovalbumin (OVA) sensitization that showed anaphylactic reaction in mice. Lung histology of OVA-sensitized mice showed narrowing of airway and large eosinophilic infiltration, whereas native and GM Brassica extract showed normal airway.. Genetically modified mustard with the codA gene possessed allergenicity similar to that of native mustard and no enhancement of IgE binding was observed due to genetic manipulation.

Topics: Adolescent; Adult; Alcohol Oxidoreductases; Animals; Arthrobacter; Female; Food Hypersensitivity; Food, Genetically Modified; Humans; Immunoglobulin E; Lung; Male; Mice; Mice, Inbred BALB C; Mustard Plant; Ovalbumin; Plants, Genetically Modified; Sequence Homology, Amino Acid

To valuate the intestinal mucosal secretary IgA (sIgA) responses to the ovalbumin-induced allergy in mice, to provide some clues for the exploration of mechanisms and therapeutic methods in the children's food allergy.. Female BALB/c mice aged 6 weeks fed on the ovalbulmin-free diet, were randomly divided into 2 groups with 8 mice in each. The mice in group Ch were sensitized with ovalbumin (OVA) intraperitoneal injection two times and challenged intragastrically 3 times. Two days after the last challenge with oral OVA, the mice were sacrificed and the samples were collected. The mice in group Ns were given intraperitoneal and intragastrical normal saline as control. The levels of total IgA and OVA-specific IgA in the intestinal mucus of the mice were determined by ELISA; the immunohistochemical methods were adopted to observe IgA(+) plasmacytes in lamina propria (LP) and surface membrane IgA (smIgA)(+) lymphocytes in peyer's patch (PP); the IL-4 mRNA expression in LP was assessed by RT-PCR. The IL-4 mRNA expression in PP was evaluated by in situ hybridization.. After the mice in Ch group were sensitized and challenged with OVA, the levels of the total IgA and the OVA-specific IgA in mucus remarkably increased (P < 0.01 respectively), the amounts of the IgA(+) plasmacytes in LP and the smIgA(+) lymphocytes in PP significantly increased (P < 0.01 respectively); a significantly positive correlation was found among the total IgA levels, the OVA-specific IgA levels, the IgA(+) plasmacyte counts in LP and the smIgA(+) lymphocyte counts in PP (P < 0.01 respectively); the mRNA expressions of IL-4 in LP and in PP were significantly augmented (P < 0.01 respectively); significantly positive correlations were found either between the IL-4 mRNA expression and the IgA(+) plasmacyte counts in LP (P < 0.01) or between the IL-4 mRNA expression and the smIgA(+) lymphocyte counts in PP (P < 0.01).. The intestinal mucosal sIgA responses are abnormally augmented in the ovalbumine-induced allergic mice, which may be partly due to the increased expression of IL-4 mRNA in gut.

Topics: Animals; Disease Models, Animal; Enzyme-Linked Immunosorbent Assay; Female; Food Hypersensitivity; Immunoglobulin A, Secretory; Immunohistochemistry; Interleukin-4; Intestinal Mucosa; Mice; Mice, Inbred BALB C; Ovalbumin; Peyer's Patches; Random Allocation; RNA, Messenger

This study sought to determine whether oral tolerance to ovalbumin (OVA), responsible for food allergy, is affected by different amounts of protein intake. For this, 6-wk-old BALB/c mice were fed with low protein (5%, LP), normal protein (20%, NP) and high protein (40%, HP) diets, orally given either OVA (OVA-fed) or water (Water-fed) for 4 d, and then immunized intraperitoneally twice at a 3-wk interval with alum-precipitated OVA. After the last immunization, sera were collected to measure total and OVA-specific IgE by enzyme assay (ELISA). Splenocytes were cultured and stimulated with concanavalin A (Con A), lipopolysaccharide (LPS) or OVA and assayed for 3H-thymidine incorporation. The culture supernatants from their splenocytes stimulated with OVA were analyzed for interleukin (IL)-4, interferon (IFN)-gamma, and IL-12. Total IgE was significantly higher in OVA-fed HP groups as compared to NP and LP groups (p<0.05). The highest and the lowest OVA-specific IgE were observed in HP and LP diet groups, respectively (p<0.05). OVA-fed mice receiving the LP diet demonstrated significantly lower IL-4 as compared to the other two groups (p<0.05), while IFN-gamma was significantly higher in the LP compared to the HP group (p<0.05). Levels of IL-12 did not differ among the OVA-fed groups. Splenocytes of OVA-fed mice kept on the LP and HP diet showed significant impairment of proliferation to OVA as compared to the NP group (p<0.01). Proliferation against Con A was impaired in the LP group compared to the NP group (p<0.05) but not in Water-fed groups. However, it was higher against LPS in the HP than the LP group (p<0.05) both in Water-fed and OVA-fed animals. All these findings indicate that established oral tolerance to OVA is clearly affected by the amount of protein diet. They support the suggestion that dietary protein plays an important role(s) in IgE-mediated food allergies.

Topics: Animals; Body Weight; Cell Proliferation; Cytokines; Dietary Proteins; Eating; Food Hypersensitivity; Immune Tolerance; Immunoglobulin E; Lymphocytes; Mice; Mice, Inbred BALB C; Ovalbumin; Spleen

Gastrointestinal allergy often precedes or coexists with respiratory allergy.. We hypothesized that established experimental gastrointestinal allergy would prime for the development of allergic respiratory responses.. BALB/c mice were sensitized with ovalbumin (OVA) in the presence of aluminum potassium sulfate and then subjected to intragastric saline or OVA challenges. After the development of allergen-induced gastrointestinal allergy, mice were intranasally exposed to either saline, OVA, or a neoaeroallergen house dust mite (HDM) extract. Airway inflammation (eg, bronchoalveolar lavage fluid cellularity, cytokine levels, and OVA-specific antibody levels) and airway responsiveness to methacholine exposure were assessed after intranasal allergen exposure.. A single intranasal exposure to OVA induced significantly more airway inflammation in intragastric OVA-challenged mice compared with that seen in intragastric saline-treated mice. Kinetic analysis revealed that the observed amplification of lung inflammation was sustained for up to 12 days after the last intragastric OVA challenge after resolution of blood eosinophilia. When mice with gastrointestinal allergy were repeatedly challenged with HDM in the respiratory tract, they experienced enhanced airway inflammation, including bronchoalveolar lavage fluid eosinophilia and increased IL-13 levels.. Taken together, our results demonstrate that OVA-induced gastrointestinal allergy enhances not only allergic airway responses to OVA but also to HDM, an unrelated aeroallergen.. Experimental gastrointestinal allergy primes for responses to allergens in the respiratory tract, enhancing antigen-specific antibody and T(H)2 cytokine production, airway inflammation, and airway hyperresponsiveness.

Topics: Allergens; Animals; Antigens, Dermatophagoides; Asthma; Bronchial Hyperreactivity; Bronchoalveolar Lavage Fluid; Eosinophilia; Female; Food Hypersensitivity; Immunoglobulin A; Immunoglobulin E; Immunoglobulin G; Male; Mice; Mice, Inbred BALB C; Ovalbumin; Pneumonia; Respiratory Hypersensitivity

Recent investigations have shown that proteins, including Bet v 1a, are nitrated by exposure to polluted urban air. We have investigated immunogenic and allergenic properties of in vitro nitrated allergens in in vivo models.. Untreated and nitrated samples of ovalbumin or Bet v 1a were compared for their ability to stimulate proliferation and cytokine secretion in splenocytes from DO11.10 or from sensitized BALB/c mice, and for their ability to induce specific immunoglobulin (Ig)G1, IgG2a and IgE in sensitized mice. Additionally, sera from birch pollen-allergic individuals were analysed for IgE and IgG specific for nitrated Bet v 1a.. Upon splenocyte stimulation with nitrated as compared with unmodified allergens, proliferation as well as interleukin 5 and interferon-gamma production were enhanced. Sera of mice sensitized with nitrated allergens showed elevated levels of specific IgE, IgG1 and IgG2a, compared with sera from mice sensitized with unmodified allergens. Moreover, cross-reactivity of antibodies against unrelated, nitrated allergens was observed in mice. We also found higher amounts of functional, specific IgE against nitrated than against untreated Bet v 1a in sera from birch pollen-allergic patients.. Our findings suggest that nitration enhances allergic responses, which may contribute to an increased prevalence of allergic diseases in polluted urban environments.

Topics: Allergens; Animals; Antigens, Plant; Cell Proliferation; Female; Food Hypersensitivity; Mice; Mice, Inbred BALB C; Ovalbumin; Plant Proteins; Protein Processing, Post-Translational; Spleen; Tetranitromethane; Tyrosine

The aim of the present study was to test a prebiotic-phytotherapic compound in an experimental model of oral allergenicity.. Antigen-specific immunoglobulin E (IgE) elevated mice were prepared by injecting them intraperitoneally with 10 microg of ovalbumin. Subsequently, the mice were exposed to ovalbumin solution intranasally and blood samples were obtained on weekly intervals for 4 weeks to measure serum-ovalbumin-specific IgE and total immunoglobulin G. Mice with high titers of ovalbumin-IgE were intragastrically administered with 0.3 mL of phosphate buffered solution containing either 20 mg of ovalbumin, the same solution with 5 mL of milk, or 20 mg milk added to prebiotic-phytocompound.. Ovalbumin administration caused a significant increase of plasma ovalbumin concentration in sensitized mice while prebiotic-phytocompound-supplemented mice showed a significantly reduced peak value (P < 0.05). Prebiotic-phytocompound added to milk exerted a significant effect in lowering the ovalbumin-IgE level and the total immunoglobulin G level as compared to control plain milk (P < 0.01).. This study provides a rationale basis for a feasible non-pharmacological therapeutic strategy in food allergen hypersensitivity syndromes.

Topics: Animals; Food Hypersensitivity; Immunoglobulin G; Lactobacillus; Male; Mice; Milk; Ovalbumin; Phytotherapy; Probiotics

We have already reported that antigen-specific IgG1 antibody production in WBB6F1-W/Wv (W/Wv) mice after oral administration of ovalbumin (OVA) was extremely high. Active systemic anaphylaxis (ASA) was induced in these mice after intraperitoneal (i.p.) administration of OVA, and Th2-dominant helper T-cell activation occurred. In this study, we examined the effect of CpG oligodeoxynucleotide (ODN) conjugation of OVA on oral immunization of W/Wv mice.. W/Wv mice were sensitized by administration of 0.1 mg OVA or CpG ODN-OVA by gavage every day for 4 weeks, and the serum titers of OVA-specific IgG1, IgE, and IgG2a antibody were determined. ASA was induced by i.p. injection of OVA, and the changes in body temperature were monitored. In vitro production of Th1- and Th2- type cytokines by splenocytes re-stimulated with antigen was also measured.. The antigen-specific IgG1 antibody titer in the CpG ODN-OVA-sensitized W/Wv mice was lower than in the OVA-sensitized group, but the IgG2a titer was higher. ASA was not induced by i.p. OVA challenge. There were significant increases in the production of Th1-type cytokine (IFN-gamma) by splenocytes in the CpG ODN-OVA-sensitized mice, but the Th2-type cytokine (IL-4) level in the splenocyte culture medium was lower.. These results indicated that oral administration of CpG ODN-OVA conjugate significantly induced antigen-specific Th1 responses and reduced Th2 responses (allergic reactions) on re-stimulation. These findings suggest that CpG ODN-antigen conjugate may be useful as an oral vaccine.

Topics: Administration, Oral; Animals; Cytokines; Female; Food Hypersensitivity; Humans; Immunoglobulin G; Immunoglobulins; Mice; Models, Animal; Oligodeoxyribonucleotides; Ovalbumin; Vaccines, DNA

Studies of the pathological mechanisms of food allergy have been impeded by the lack of relevant animal models. The purpose of this study was to develop a physiological model of food allergy that was not dependent on immunostimulatory adjuvants.. Balb/c mice were epicutaneously sensitized four times at varying intervals over a 22-day period, and challenged orally from day 40, 6 times every 1-3 days with either saline or ovalbumin.. After sensitization (day 35) but before the oral challenges, the ovalbumin-sensitized groups showed increased specific IgE and IgG1 production when compared with the sham-sensitized groups. Mucosal mast cell protease-1 (MMCP-1) was undetectable in serum before the intragastric challenge. MMCP-1 concentrations were increased after the first ovalbumin dose, solely in the ovalbumin-sensitized and -challenged group. After the challenge period, the mean serum MMCP-1 concentration increased from an undetectable level in controls to an over 44-fold level in the ovalbumin-sensitized and -challenged mice. In this group, MMCP-1-positive cells were present in the small intestine and expressions of IFN-gamma and CXCL-9 mRNA were decreased in the ileum, suggesting an impaired Th-1-type response. Within one hour of the last ovalbumin challenge, 5 out of 6 mice developed diarrhea in the ovalbumin-sensitized and -challenged group, but there was no diarrhea in the other groups.. A murine model of food allergy based on sensitization via epicutaneous exposure to allergen without immunostimulatory adjuvants was developed. Effective production of MMCP-1 together with specific IgE and IgG1 suggests a breakdown in oral tolerance to the allergen. Intragastric challenges were accompanied by mast cell-dependent immunopathological changes and diarrhea.

Topics: Animals; Chymases; Diarrhea; Disease Models, Animal; Female; Food Hypersensitivity; Immunization; Immunoglobulin E; Immunoglobulin G; Immunohistochemistry; Mice; Mice, Inbred BALB C; Ovalbumin

An increase in plasma ovalbumin concentrations after intragastric administration of ovalbumin was suppressed by concomitant freeze-dried kefir in BALB/c mice. Serum levels of ovalbumin-specific immunoglobulin G and proliferation of splenic mononuclear cells in mice immunized orally with ovalbumin were suppressed by feeding freeze-dried kefir. We propose that kefir reduces intestinal permeation of food antigen, which contributes to suppression of oral sensitization.

Topics: Administration, Oral; Animals; Antigens; Cultured Milk Products; Female; Food Hypersensitivity; Freeze Drying; Intestinal Absorption; Mice; Mice, Inbred BALB C; Ovalbumin

Undiagnosed food allergies have been proposed as possible causes of promoting and perpetuating irritable bowel syndrome . Our aim was to find out if sensitization could induce chronic functional motor disturbances in the intestine and the mechanisms implicated. Rats were sensitized to ovalbumin (OVA) following three hypersensitivity induction protocols, two parenteral and one oral. Rat mast cell protease II (RMCP II) release in response to OVA challenge and immunoglobulin E (IgE) concentration were measured in serum. At least 1 week after challenge, small intestinal motility was evaluated using strain gauges. Intestinal tissue samples from orally sensitized rats were checked for in vitro stimulation with OVA. Mucosal mast cells were counted from duodenum sections. All sensitized rats showed intestinal hypermotility. Only rats sensitized by parenteral procedure showed an increase in RMCP II after OVA challenge in serum. IgEs increased only in the Bordetella pertussis sensitized group. Small intestine sections from orally sensitized rats released more RMCP II than sections from control rats. All sensitized rats showed an increase in the number of mucosal mast cells in duodenum. In conclusion, hypersensitivity to food proteins induces chronic motor alteration that persists long after antigen challenge and an excited/activated state of sensitized mucosal mast cells.

Topics: Animals; Bordetella pertussis; Cholecystokinin; Duodenum; Food Hypersensitivity; Gastrointestinal Motility; Ileum; Immunoglobulin E; Immunohistochemistry; Intestinal Diseases; Intestinal Mucosa; Intestine, Small; Male; Mast Cells; Muscle Tonus; Ovalbumin; Rats; Rats, Sprague-Dawley; Serine Endopeptidases

We have already reported that WBB6F1-W/W(v) (W/W(v)) mice, which have mutations in the c-kit gene, are highly susceptible to oral sensitization, and that the proportion of TCRgammadelta-T cells among the intraepithelial lymphocytes (IELs) (gammadelta-IELs) of W/W(v) is much lower than in congenic wild-type (+/+) mice. In this study we examined an inhibitory role of gammadelta-IELs in oral sensitization using two different methods. First, wild-type (+/+) mice were sensitized by oral administration of 1.0 mg ovalbumin (OVA) by gavage every day for 9 weeks after anti-TCRgammadelta antibody treatment 4 times. The treatment resulted in an enhanced OVA-specific IgG1 antibody production, active systemic anaphylaxis (ASA), and Th2-dominant cytokine production. Next, W/W(v) mice whose bone marrow cells were reconstituted from C57BL/6J mice for 5 months were sensitized by oral administration of OVA. The OVA-specific IgG1 antibody titer in the bone marrow-reconstituted W/W(v) mice was neither significantly enhanced, nor ASA was induced. The proportion of gammadelta-IELs in the reconstituted mice was much higher than that in the untreated W/W(v) mice. The above findings suggest that the decrease or increase in number of gammadelta-IELs enhances or decreases oral sensitization respectively. These results show that gammadelta-IELs have an important role in the oral tolerance to food antigens.

Topics: Administration, Oral; Animals; Antibodies; Antigens; Body Temperature; Bone Marrow Cells; Female; Food Hypersensitivity; Immunity, Mucosal; Mice; Mice, Inbred Strains; Mutation; Ovalbumin; Proto-Oncogene Proteins c-kit; Receptors, Antigen, T-Cell, gamma-delta; T-Lymphocytes; Time Factors

Some epidemiological or association studies suggest that transforming growth factor-beta (TGF-beta) in breast milk may be a decisive factor in diminishing the risk of allergic diseases during infancy. The observations have prompted us to investigate whether TGF-beta, when taken orally, can affect allergic immune responses. Repeated high-dose ovalbumin peptide (OVA) feeding was previously reported to induce OVA-specific IgE production and an anaphylactic reaction after intravenous challenge of OVA in OVA-TCR transgenic mice, which might represent a model for food allergy. By using this model, we showed here that oral administration of high-dose TGF-beta simultaneously with OVA feeding significantly inhibited the OVA-specific IgE elevation and anaphylactic reaction in OVA-TCR transgenic DO11.10 mice. These effects were associated with suppression of OVA-specific IL-4 production and GATA-3 expression and with up-regulation of IFN-gamma production and T-bet expression by splenocytes. Intra-peritoneal injection of anti-TGF-beta-neutralizing antibody abolished the inhibitory effects of orally administered TGF-beta on the serum IgE response and anaphylactic reaction after OVA feeding in DO11.10 mice. Interestingly, oral administration of high-dose TGF-beta suppressed activation-induced T cell death induced by OVA feeding in DO11.10 mice. We thus conclude that TGF-beta, when taken orally at high dose, has the capacity to modulate a food allergy-related reaction, at least in part, through its systemic activity.

Topics: Administration, Oral; Anaphylaxis; Animals; Antigens, Differentiation, T-Lymphocyte; Apoptosis; CD4-Positive T-Lymphocytes; Cytokines; Fas Ligand Protein; Food Hypersensitivity; Immunoglobulin E; Immunoglobulin G; Injections, Subcutaneous; Membrane Glycoproteins; Mice; Mice, Inbred BALB C; Mice, Transgenic; Ovalbumin; Spleen; T-Lymphocyte Subsets; Transforming Growth Factor beta; Tumor Necrosis Factors

This study was to develop a suitable model for the investigation for food allergy.. BALB/c mice were dosed by intraperitoneally with Ovalbumin, Beef serum albumin, Trypsin inhibitor and Potato acid phosphatase respectively (0.25ml 20mg/mnl) on day 0 and again on day 7. Control group was dosed with PBS. Sera form individual animals were analysed for specific IgE and Passive cutaneous anaphylaxis tests. Additionally, the level of histamine in plasma were detected.. The high titres of specific IgE (1: 32) could be provoked in test groups compared with control group. In addition, the level of histamine in plasma of test groups was higher than that in the control group. But there was no statistical significance between group food allergen and group Potato acid phosphatase.. Although allergic action of BALB/c mice could be provoked, the situation of the allergic action of BALB/c mice to the proteins was very different with the human being. The BALB/c mice could not be a suitable model for the investigation for food allergy.

Topics: Acid Phosphatase; Animals; Disease Models, Animal; Female; Food Hypersensitivity; Histamine; Immunoglobulin E; Male; Mice; Mice, Inbred BALB C; Ovalbumin; Serum Albumin, Bovine; Trypsin Inhibitors

Rabbit MAC-1 receptor, homologue to human CD11b is present in macrophages. The aim of the study was to determine quantitative and distributive modifications of CD11b-positive cells that participate in immune response at rectal mucosa, in an animal model of mucosal immunity. New Zealand rabbits were divided into three groups. G1: control; G2: ovalbumin (OVA) sensitized; G3: OVA-senstitized and rectal challenged. Animals were subcutaneously sensitized twice with 70 microg OVA and 30 ml aluminium hydroxide in 2 ml saline solution. Rectal challenge was developed with a solution of 50 mg OVA in 5 ml saline solution. Sensitized groups (G2 and G3) showed a positive PCA (Passive Cutaneous Anaphylaxis) at 1/160 fold dilutions. In G3 we observed a patchy mucosal edema, lymphangiectasis and eosinophil leucocyte infiltration. Cells were counted as the number of cells per high power field. G1: 9.64 (SE 0.22); G2: 18.10 (SE 0.09) and G3: 23.60 (SE 0.29). (G2 vs G1 p < 0.001; G3 vs G1 p < 0.001; G3 vs G2 p < 0.001). We conclude that there is a close relationship between the food antigen OVA penetration (after challenge) and the increase of CD11b positive cells in rectal mucosa. This fact could be due to the cellular influx to the inflammatory site by the action of chemotactic factors released after challenge.

Topics: Animals; CD11b Antigen; Cell Count; Disease Models, Animal; Food Hypersensitivity; Immunity, Mucosal; Immunization; Immunoglobulin E; Macrophage-1 Antigen; Male; Mucous Membrane; Ovalbumin; Rabbits; Rectum

This study investigated whether orally administered probiotic bacteria (Bifidobacterium bifidum and Lactobacillus casei) and a gram-negative bacterium (Escherichia coli) function as allergic immune modulators to prevent food allergy, according to the hygiene hypothesis. C3H/HeJ mice were sensitized with ovalbumin (OVA) and cholera toxin for 5 weeks. After sensitization, the OVA-induced mice that were not treated with bacteria had significantly increased levels of OVA-specific IgE, total IgE, and IgG1 in sera, as well as scab-covered tails. In comparison, groups treated with B. bifidum BGN4 (BGN4), L. casei 911 (L. casei), or Escherichia coli MC4100 (E. coli) had decreased levels of OVA-specific IgE, total IgE, and IgG1, and decreased levels of mast cell degranulation and tail scabs. OVA-specific IgA levels were decreased in BGN4- and L. casei-treated groups. In conclusion, administration of E. coli, BGN4, or L. casei decreased the OVA-induced allergy response. However, a normal increase in body weight was inhibited in the E. coli-treated mice and in the montreated mice groups during allergy sensitization. Thus, BGN4 and L. casei appear to be useful probiotic bacteria for the prevention of allergy.

Topics: Administration, Oral; Animals; Bifidobacterium; Body Weight; Cell Degranulation; Cholera Toxin; Disease Models, Animal; Escherichia coli; Female; Food Hypersensitivity; Immunity, Mucosal; Immunoglobulin A, Secretory; Immunoglobulin E; Immunoglobulin G; Lacticaseibacillus casei; Mast Cells; Mice; Mice, Inbred C3H; Ovalbumin; Probiotics

The incidence of type I allergic disorders has been increasing worldwide, particularly, the hypersensitivity to food. We first showed that apple condensed tannin (ACT) intake would inhibit the development of the oral sensitization and that the inhibition could correlate with the rise in the population of TCR(gamma)delta-T cells in the intestinal intraepithelial lymphocytes (IEL) using W/W(V) mice and B10A mice which were ovalbumin (OVA)-orally sensitized. Serum OVA-specific immunoglobulin E and immunoglobulin G1 titers in the OVA-orally sensitized W/W(V) and B10A mice ad libitium fed ACT were extremely inhibited compared to those of the control. The ACT intakes of OVA-sensitized W/W(V) and B10A mice inhibited the immediate reduction of the body temperature or the rise in serum histamine induced by active systemic anaphylaxis. The proportions of the TCR(gamma)delta-T cells in the IEL of the OVA-orally sensitized W/W(V) and B10A mice ad libitium fed ACT were significantly greater than that in the controls. Furthermore, ACT feeding by itself could induce the rise in the percentage of the TCR(gamma)delta-T cells among the IEL of the W/W(V) and B10A mice. This suggests that the ACT intake may prevent the development of food allergies and this effect could be correlated with the rise in the percentage of TCR(gamma)delta-T cells among the IEL.

Topics: Allergens; Animals; Body Temperature; Body Weight; Disease Models, Animal; Female; Flavonoids; Food Hypersensitivity; Histamine; Immunoglobulin E; Immunoglobulin G; Intestinal Mucosa; Malus; Mice; Mice, Inbred Strains; Ovalbumin; Phenols; Polyphenols; Proanthocyanidins; Receptors, Antigen, T-Cell, gamma-delta; T-Lymphocytes

To develop a Brown Norway (BN) rat model to determine the potential allergenicity of novel proteins in genetically modified food.. The allergenicity of different proteins were compared, including ovalbumin (OVA), a potent respiratory and food allergen, bovine serum albumin (BSA), a protein that is considered to have a lesser allergenic potential, and potato acid phosphatase (PAP), a non-allergenic protein when administered to BN rats via different routes of exposure (intraperitoneally or by gavage). IgG and IgE antibody responses were determined by ELISA and PCA, respectively. An immunoassay kit was used to determine the plasma histamine level. In addition, possible systemic effect of allergens was investigated by monitoring blood pressure.. OVA provoked very vigorous protein-specific IgG and IgE responses, low grade protein-specific IgG and IgE responses were elicited by BSA, while by neither route did PAP elicit anything. In either routes of exposure, plasma histamine level in BN rats sensitized with OVA was higher than that of BSA or PAP. In addition, an oral challenge with BSA and PAP did not induce any effect on blood pressure, while a temporary drop in systolic blood pressure in few animals of each routes of exposure was found by an oral challenge with OVA.. BN rat model might be a useful and predictive animal model to study the potential allergenicity of novel food proteins.

Topics: Acid Phosphatase; Animals; Disease Models, Animal; Food Hypersensitivity; Food, Genetically Modified; Male; Ovalbumin; Rats; Rats, Inbred BN; Serum Albumin, Bovine; Solanum tuberosum

The aim is to determine immunopathological modifications in rectal mucosa from rabbit after local challenge in sensitized animals with ovalbumin (OVA).. Thirty rabbits divided into three groups: G1: normal, G2: subcutaneously OVA sensitized, G3: sensitized, locally OVA challenged and sampled 4 hours after challenge. Specific anti-OVA IgE levels were evaluated by passive cutaneous anaphylaxis test (PCA). In each group 200 high microscopical power fields (HPF) were counted. Results were expressed as arithmetic mean and SE. Statistical analysis was made using Student t test. Anti-CD4, CD5, micro chain, CD25 and RLA II monoclonal antibodies were used. Avidin biotin horseradish peroxidase system was used.. CD 4: G1: 8.3 +/- 0.06; G2: 13.4 +/- 0.08 and G3: 8.25 +/- 0.06. CD 5: G1: 7.3 +/- 0.05; G2: 9.4 +/- 0.05 and G3: 11.3 +/- 0.06. CD 25: G1: 13 +/- 0.08; G2: 15.1 +/- 0.13 and G3: 25.5 +/- 0.15. mu chain: G1: 10.4 +/- 0.06; G2: 3.8 +/- 0.02 and G3: 6.0 +/- 0.10. RLA II (DR): G1: 11.6 +/- 0.05; G2: 19.2 +/- 0.09 and G3: 19.1 +/- 0.11. In all cases, experimental groups (G2 and G3) presented statistical significant differences vs. control group (G1) (p < 0.001).. Interleukin-2 receptor (CD25+ cells) increase in experimental groups. Cells expressing micro chain decreased in G2 and G3 likely due to activation of B cells and subsequent expression of other immunoglobulin chains in cell surface. RLA II expression is higher in G2 and G3. This receptor is considered an activation marker expressed by macrophages, T and B cells. We conclude that obtained data are important to elucidate immunopathology of local anaphylactic reaction in rectal mucosa from systemic sensitized animals.

Topics: Allergens; Animals; Biomarkers; Disease Models, Animal; Food Hypersensitivity; Intestinal Mucosa; Ovalbumin; Rabbits; Rectum

The contribution of different T cell subsets to the overall measured cytokine response to food allergens is largely unexplored.. The patterns of cytokine production of peripheral blood-derived T cells after allergen stimulation were studied in 22 children with multiple food allergies and in 20 non-allergic children as controls, using flow cytometry.. Proportions of T cells of food-sensitized children spontaneously secreting IFN-gamma and IL-10 (without antigen stimulation) were lower than non-atopic children and adult controls (P

Topics: Allergens; Cells, Cultured; Child; Child, Preschool; Cytokines; Dendritic Cells; Female; Food Hypersensitivity; Humans; Immunoglobulin E; Interferon-gamma; Interleukin-10; Interleukin-2; Interleukin-4; Lactoglobulins; Leukocyte Common Antigens; Male; Ovalbumin; Peanut Hypersensitivity; T-Lymphocytes

To evaluate the immunoregulatory effects of the Lonicera water extract in the ovalbulmin (OVA)-sensitized BALB/c mice and to explore feasibility of treating food allergy with the traditional Chinese medicine (TCM).. Forty female BALB/c mice aged 6 weeks fed with ovalbulmin-free feed, were randomly divided into 5 groups with 8 mice in each. Four groups were sensitized with OVA intraperitoneally two times and challenged intragastrically four times. Groups H, M and L were treated respectively with high (100 mg/100 ml), medium (50 mg/100ml) and low (25 mg/100 ml) concentration of the Lonicera water extract at a dose of 0.3 ml/10 g body weight just 4 hours after the first challenge and then twice daily for 10 consecutive days. The mice in group Ch were used as positive control and were sensitized intraperitoneally and treated with normal saline solution intragastrically daily. The mice in NS group were used as negative control without sensitization and challenge. Just 1 hour after the last challenge, the mice in each group were sacrificed and specimens of jejunum were taken. Histological examinations on the jejunum specimens were performed after either HE or toluidine blue staining, the levels of histamine in gut of the mice were assayed with a fluorescent method; the IFN-gamma and IL-4 production in peripheral lymph node mononuclear cell (PLNMC) and the OVA-specific IgE levels in serum were measured by using ELISA; the mRNA expression of IL-12p40 in PLNMC of the mice was evaluated by RT-PCR; the footpad swelling reactions were assessed for the OVA-induced delayed hypersensitivity.. (1) The inflammatory reactions were significantly inhibited in the mice of group H and M; the accumulated and degranulated mast cells in lamina propria were significantly reduced in the mice by gavage with 100% or 50% of the Lonicera extract, concomitant with the increased percentage of the intact mast cells. (2) The release of histamine in gut in the mice of group H and M was significantly reduced. (3) Either the IL-4 production and the ratio of IL-4/IFN-gamma in PLNMC or the IFN-gamma generation was significantly reduced in group H and M. (4) IL-12p40 mRNA expression in PLNMC was significantly reduced in group H and M. (5) The levels of OVA-specific IgE in serum were reduced in the mice of group H and M. (6) The footpad swelling reactions induced in the allergic mice were significantly inhibited after giving the Lonicera extract of the three different concentrations.. The Lonicera extract showed significant immunoregulatory effects in OVA-induced allergic mice model in this study. Lonicera extract may be of potential research value in treatment of both IgE and none IgE mediated food allergy.

Topics: Animals; Female; Food Hypersensitivity; Histamine; Immunoglobulin E; Interferon-gamma; Interleukin-12 Subunit p40; Interleukin-4; Jejunum; Lonicera; Mast Cells; Mice; Mice, Inbred BALB C; Ovalbumin; Plant Extracts

Few models have described a chronic food allergy with morphological changes in the intestinal mucosa. Here we established an ovalbumin (OVA)-induced, cell-mediated, allergic rat model and examined lymphocyte migration in the gut. Brown Norway rats were intraperitoneally sensitized to OVA and then given 10 mg OVA/day by gastric intubation for 6 wk. Lymphocyte subsets and adhesion molecules were examined immunohistochemically, and the migration of T lymphocytes to microvessels of Peyer's patches and villus mucosa was observed by using an intravital microscope. Serum OVA-specific IgG and IgE levels were increased in animals repeatedly exposed to OVA. Significant villus atrophy and increased crypt depth was accompanied by increased infiltration of T lymphocytes in the small intestinal mucosa of the group given OVA. Expression of rat mast cell protease II and of mucosal addressin cell adhesion molecule-1 (MAdCAM-1) was also increased in these groups. The administration of anti-MAdCAM-1 antibody significantly attenuated the OVA-induced changes in the mucosal architecture and in CD4 T lymphocyte infiltration. Intravital observation demonstrated that in rats with a chronic allergy, T lymphocytes significantly accumulated in villus microvessels as well as in Peyer's patches via a MAdCAM-1-dependent process. Our model of chronic food allergy revealed that lymphocyte migration was increased with MAdCAM-1 upregulation.

Topics: Animals; Antibodies; Cell Adhesion Molecules; Cell Movement; Chronic Disease; Diet; Food Hypersensitivity; Hypersensitivity, Delayed; Immunoglobulins; Intestinal Mucosa; Intestine, Small; Lymphocyte Subsets; Lymphocytes; Male; Microvilli; Mucoproteins; Ovalbumin; Peyer's Patches; Rats; Rats, Inbred BN; Serine Endopeptidases; Venules

Tryptophan was isolated from rat feces as an active compound against ovalbumin permeation in an in vitro Caco-2 cell model. Tryptophan dose-dependently inhibited ovalbumin permeation with accompanying increase in transepithelial electric resistance, and its inhibitory activity reached a plateau at 10 mM. Brown Norway rats were sensitized by intragastric administration of ovalbumin together with or without tryptophan. Antibody levels specific to ovalbumin in the sera and proliferative responses of spleen mononuclear cells to ovalbumin were significantly lower in rats administered ovalbumin plus tryptophan than those administered ovalbumin alone. These results suggest that tryptophan suppresses oral sensitization to ovalbumin, probably via suppression of ovalbumin absorption from the intestinal tract.

Topics: Administration, Oral; Allergens; Animals; Antibodies; Caco-2 Cells; Dose-Response Relationship, Drug; Feces; Food Hypersensitivity; Humans; Immunization; Intestines; Male; Ovalbumin; Permeability; Rats; Rats, Inbred BN; Rats, Wistar; Spleen; Tryptophan

Food allergy is an important and common health issue, and there is a need to identify and characterize the sensitizing mechanisms. One of the common causes of food allergy is ovalbumin (OVA), a dietary antigen from eggs. We hypothesized that OVA-induced food allergy in the gut involves the activation of the chemokine regulated on activation, normal T cell expressed and secreted (RANTES), which then recruits eosinophils to lesioned tissue. The purpose of this study was to clarify whether RANTES expression correlates with eosinophil infiltration in the gut of OVA-sensitized BALB/c mice in response to oral OVA challenge. BALB/c mice were immunized with OVA 1 microg and sensitized after 2 weeks by intragastric administration of OVA. Sensitization to the oral OVA challenge was analyzed by examining eosinophil infiltration into the gut tissue (immunohistochemistry), mucosal eosinophil cationic protein (ECP) concentration, and RANTES mRNA expression (reverse-transcriptase polymerase chain reaction and Southern blotting) at 3, 6, 12, and 24 h after the challenge. There was marked edema of the intestinal villi, and eosinophil infiltration to the lamina propria peaked at 6 h in OVA-sensitized mice. RANTES mRNA expression peaked at 3 h and 6 h and declined thereafter. The expression of RANTES mRNA in the allergic mice was much higher than in the nonallergic, normal, or unsensitized control mice. Tissue eosinophilia and intestinal ECP levels were significantly correlated with the RANTES mRNA level. We conclude that RANTES may play a central role in the pathogenesis of food-mediated gastrointestinal allergy.

Topics: Animals; Chemokine CCL5; Disease Models, Animal; Edema; Eggs; Eosinophils; Female; Food Hypersensitivity; Immunoglobulin E; Intestinal Mucosa; Jejunum; Leukocyte Count; Mice; Mice, Inbred BALB C; Ovalbumin; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger

The purpose of the presented experiments was to study the possibility of using the Brown Norway rat as a model for food allergy in our laboratory. Specific serum IgE against ovalbumin (OVA) was induced after dosing male and female Brown Norway rats daily by gavage for 35 days. The influence of various preparations of allergen: OVA grade II, OVA grade V, and fresh egg white, age (4 versus 8 weeks), dosing volumes, and animal suppliers was studied. A general finding was that females had statistically significantly higher specific IgE and IgG titres and number of responders than males. Egg white preparation, age, dosing volume, and animal supplier did not statistically significantly influence the median IgE and IgG titres and number of responders. The difference between immune responses in males and females could not be attributed to variations in daily intake of OVA or exposure via the lung. In our hands, the oral Brown rat food allergy model gives rise to a moderate number of IgE responders, 13-38 and 38-75% in males and females, respectively. For further experiments with this model in our laboratory, females seem the sex of choice.

Topics: Aging; Allergens; Animals; Disease Models, Animal; Dose-Response Relationship, Drug; Enzyme-Linked Immunosorbent Assay; Female; Food Hypersensitivity; Immunoglobulin E; Immunoglobulin G; Male; Ovalbumin; Rats; Rats, Inbred BN; Sex Characteristics

Protein allergenicity was investigated in BN rat model.. OVA or BSA was administered i.p. (1 mg/ml) or orally (10 mg/ml) to male BN rats for 6 weeks. In different intervals, blood samples were obtained from the orbital plexus for PCA and histamine determination. In addition, blood pressure of rats in OVA-treated group and control group was measured.. Relatively high titres (1/32-1/128) IgE responses were provoked by administration of OVA, while BSA stimulated only low titres (1/2-1/4) IgE. Compared with the controls, the plasma histamine concentrations of OVA groups were higher. An oral challenge with OVA did not induce a clear effect on blood pressure in the majority of rats. However, two rats demonstrated a temporary drop in blood pressure.. BN rat model may be a suitable animal model to determine protein allergenicity.

Topics: Animals; Dietary Proteins; Food Hypersensitivity; Histamine; Immunoglobulin E; Male; Ovalbumin; Passive Cutaneous Anaphylaxis; Random Allocation; Rats; Rats, Inbred BN; Serum Albumin, Bovine

There is increasing evidence supporting the notion that brain-gut communication is crucial for the manifestation of functional gastrointestinal (GI) disorders. Employing denervation by neonatal capsaicin treatment, we investigated here the role of unmyelinated C-fibers in food allergy signaling in the brain. We found that 90 min after oral ovalbumin (OVA) challenge, allergic mice present increased c-fos expression in emotionality-related brain areas such as the paraventricular nucleus of the hypothalamus (PVN) and the central nucleus of the amygdala (CeA). Food allergy also induced enhanced Fos immunoreactivity in the nucleus of tractus solitarii (NTS) of OVA-immunized animals. We also show that while the degree of Fos staining in the NTS of allergic mice was only diminished by neonatal capsaicin, it was completely blocked in the PVN. However, capsaicin did not modify food allergy-induced c-fos expression in the CeA. In conclusion, this study provides evidence showing that unmyelinated C-fibers are part of the neural pathways involved in food allergy-induced activation of specific brain areas, particularly the PVN and to a lesser extent the NTS.

Topics: Analysis of Variance; Animals; Animals, Newborn; Capsaicin; Cell Count; Food Hypersensitivity; Gene Expression Regulation; Immunohistochemistry; Male; Mice; Mice, Inbred BALB C; Neural Pathways; Neurons, Afferent; Ovalbumin; Paraventricular Hypothalamic Nucleus; Proto-Oncogene Proteins c-fos; Solitary Nucleus

Food allergy is most frequently the result of IgE-mediated hypersensitivity reactions. Here, we describe a chronic model in which some of the intestinal and systemic consequences of continuous egg white solution ingestion by ovalbumin-sensitized eight-week-old BALB/c mice, 6 animals per group, of both sexes, were investigated. There was a 20% loss of body weight that began one week after antigen exposure and persisted throughout the experiment (3 weeks). The sensitization procedure induced the production of anti-ovalbumin IgG1 and IgE, which were enhanced by oral antigen exposure (129% for IgG1 and 164% for IgE, compared to sensitization values). Intestinal changes were determined by jejunum edema at 6 h (45% Evans blue extravasation) and by a significant eosinophil infiltration with a peak at 48 h. By day 21 of continuous antigen exposure, histological findings were mild, with mast cell hyperplasia (100%) and increased mucus production (483%). Altogether, our data clearly demonstrate that, although immune stimulation was persistently occurring in response to continuous oral antigen exposure, regulatory mechanisms were occurring in the intestinal mucosa, preventing overt pathology. The experimental model described here reproduces the clinical and pathological changes of mild chronic food allergy and may be useful for mechanistic studies of this common clinical condition.

Topics: Animals; Chronic Disease; Disease Models, Animal; Female; Food Hypersensitivity; Immunoglobulin E; Intestine, Small; Male; Mice; Mice, Inbred BALB C; Neutralization Tests; Ovalbumin

Egg sensitization, particularly persistent sensitization, is a risk factor for later asthma. However, little is known about accompanying IgG and subclass responses and how they might relate to asthmatic outcome.. To characterize hen's egg ovalbumin (OVA) IgG and subclass responses through the first 5 years of life in relation to duration of egg sensitization and later asthma.. The subjects (n=46) formed part of a larger cohort, born to atopic parents, who had been evaluated prospectively for the development of asthma. Egg sensitization was classified as transient (positive egg skin prick test at 1 year only) or persistent (positive skin test for at least 2 years). Plasma OVA IgG, IgG1 and IgG4 concentrations at birth (cord), 6 months, 1 and 5 years of age were measured by ELISA.. The kinetics of OVA IgG and IgG1 responses, but not IgG4, differed between egg sensitized and non-egg sensitized (NES) children. Only persistently sensitized children had a rise in OVA IgG1 concentration through the first year of life, and at 1 year of age they had significantly higher OVA IgG and IgG1 than either transiently sensitized or NES children. High OVA IgG1 was associated with later asthma: at 1 year of age, OVA IgG1 greater than 14,500 U predicted asthma with a sensitivity 64% and specificity 74%.. OVA IgG and subclass responses relate to the duration of egg sensitization. Measurement of OVA IgG1 concentration in infancy might offer a useful adjunct to identify those at an increased risk of asthma.

Topics: Antibody Specificity; Asthma; Child, Preschool; Dermatitis, Atopic; Eggs; Food Hypersensitivity; Humans; Immunoglobulin G; Infant; Ovalbumin; Prognosis; Prospective Studies; ROC Curve; Skin Tests

Oral tolerance is a specific immune unresponsiveness to food antigens to prevent hypersensitivity reactions. We investigated whether zinc deficiency affects oral tolerance. Rats were fed a control (C) or zinc-deficient (ZD) diet, or pair-fed (PF) to ZD rats for 28 d. Beginning on d 7, rats were administered ovalbumin (OVA) orally to induce tolerance, or PBS 3 times/wk, and were then immunized by OVA injection. The proliferation of mesenteric lymph node (MLN) and spleen lymphocytes after in vitro OVA stimulation and the delayed-type hypersensitivity were higher in OVA-fed ZD than in OVA-fed C rats and not different between OVA- and PBS-fed ZD rats, indicating a suppression of tolerance. Lymphocyte proliferation did not differ between PF and C rats. Expressions of cytokines involved in oral tolerance, i.e., interleukin (IL)-4, IL-10 and transforming growth factor-beta, were higher in OVA- than in PBS-fed C rats, but not in ZD rats. Apoptosis was higher in OVA- than in PBS-fed C rats but not different between OVA- and PBS-fed ZD rats. Inflammation and ulcerations that were not present in ZD rats on d 7 (ZD(7)) developed in OVA- or PBS-fed ZD rats. Compared with ZD(7) rats, tumor necrosis factor-alpha and cytokine-induced neutrophil chemoattractant were higher in OVA- and PBS-fed ZD rats, whereas interferon-gamma increased only in OVA-fed ZD rats. In conclusion, zinc deficiency suppresses oral tolerance through dysregulation of cytokine expression and lack of antigen-specific clonal deletion. We suggest that abrogation of tolerance may lead to development of mucosal inflammation and damage.

Topics: Animals; Food Hypersensitivity; Intestinal Mucosa; Jejunum; Male; Ovalbumin; Peroxidase; Rats; Rats, Sprague-Dawley; Spleen; Tumor Necrosis Factor-alpha; Zinc

Other researchers have reported that the specific immune response to subsequent antigen challenge is primed in newborn mice or rats dosed orally by gavage. We wanted to investigate if priming of a subsequent specific IgE response could be achieved by dosing newborn rats orally with ovalbumin and if this method could be used in an animal model for food allergy.. Newborn Brown Norway rats were dosed with ovalbumin in the mouth (100 microg or 6 mg). As young adults, the animals were dosed by gavage for 35 days with 1 mg ovalbumin/day or once intraperitoneally with 100 microg. Control groups were dosed by gavage or intraperitoneally but not as neonates. Additionally, young adult rats were dosed with 1 mg ovalbumin/day in the mouth for 35 days. Sera from individual animals were analysed for specific IgE and specific IgG.. In all experiments with neonatal rats the specific IgE and IgG responses were decreased compared to the control groups, however, not always reaching statistical significance. A statistical significant decrease in the specific immune response was found in young adult rats dosed in the mouth as compared to by gavage.. Dosing Brown Norway rats with ovalbumin in the mouth as neonates do not prime the specific immune response. The decrease in immune response found in our experiments when dosing newborn animals in the mouth in opposition to the priming seen by others when dosing by intragastric intubation may be explained by a dissimilar antigen presentation when dosing includes both oral mucosa and gut.

Topics: Animals; Animals, Newborn; Disease Models, Animal; Food Hypersensitivity; Immunoglobulin E; Immunoglobulin G; Ovalbumin; Rats; Rats, Inbred BN

Topics: Blood Proteins; Cardiac Surgical Procedures; Egg Proteins; Eggs; Food Hypersensitivity; Humans; Ovalbumin

Although T-cell responses to food antigens are normally inhibited either by deletion, active suppression, or both of antigen-specific T cells, T helper cells for IgE response to a food antigen still develop by unknown mechanisms in a genetically susceptible host.. We determined the site at which those IgE helper T cells develop.. We administered ovalbumin (OVA) orally to DO11.10 mice and studied CD4+ T cells in Peyer's patches, the spleen, and the liver. Helper activity for IgE response was assessed by adoptively transferring those CD4+ T cells to naive BALB/c mice, followed by systemic immunization with OVA.. OVA-specific CD4+ T cells were deleted by cell death in the liver and Peyer's patches of DO11.10 mice fed OVA. OVA-specific CD4+ T cells that survived apoptosis in the liver expressed Fas ligand and secreted IL-4, IL-10, and transforming growth factor beta(1). CD4+ T cells producing IFN-gamma were deleted in the liver by repeated feeding of OVA. On transfer of CD4+ T cells to naive mice and systemic immunization with OVA, a marked increase in OVA-specific IgE response developed only in the mice that received hepatic CD4+ T cells from OVA-fed mice, the effect of which was not observed in the recipients of hepatic CD4+ T cells deficient in IL-4. In addition, significant suppression of delayed-type hypersensitivity and IgG(1)/IgG(2a) responses to OVA was observed in the recipients of hepatic CD4+ T cells, and this suppression required Fas/Fas ligand interaction.. Together, these results suggested that a food antigen might negatively select helper T cells for IgE response to the antigen by preferential deletion of T(H)1 cells in the liver.

Topics: Administration, Oral; Adoptive Transfer; Animals; Antigens; Cells, Cultured; Clonal Deletion; Fas Ligand Protein; Food Hypersensitivity; Genes, T-Cell Receptor; Hypersensitivity, Delayed; Immunoglobulin E; Interleukin-4; Liver; Membrane Glycoproteins; Mice; Mice, Inbred BALB C; Mice, Knockout; Mice, Transgenic; Ovalbumin; Peyer's Patches; Spleen; T-Lymphocytes, Helper-Inducer; Th2 Cells

Although many authors have considered the possibility of a direct interaction between food allergy and behavioral changes, the evidence supporting this hypothesis is elusive. Here, we show that after oral ovalbumin (OVA) challenge, allergic mice present higher levels of anxiety, increased Fos expression in emotionality-related brain areas, and aversion to OVA-containing solution. Moreover, treatment with anti-IgE antibody or induction of oral tolerance abrogate both food aversion and the expression of c-fos in the central nervous system (CNS). Our findings establish a direct relationship between brain function and food allergy, thus creating a solid ground for understanding the etiology of psychological disorders in allergic patients.

Topics: Administration, Oral; Animals; Antibodies, Anti-Idiotypic; Anxiety; Avoidance Learning; Brain Chemistry; Emotions; Feeding Behavior; Food Hypersensitivity; Immunization; Immunoglobulin E; Injections, Intraperitoneal; Male; Mice; Mice, Inbred BALB C; Neurons; Ovalbumin; Proto-Oncogene Proteins c-fos

The aetiology of food allergy remains unclear. Although failure to develop or breakdown in oral tolerance has been proposed, the existence of physiologic sensitization routes other than the gastrointestinal tract cannot be excluded.. The purpose of this study is to clarify whether or not exposure to allergen through the skin can promote food allergy.. BALB/c mice were shaved on the back, and a patch impregnated with 100 micro g of ovalbumin (OVA) was applied to the dorsal skin for a 1-week period and then removed. After three courses of sensitization, OVA-specific antibodies in sera were measured, and then mice were orally challenged with 50 mg of OVA. Anaphylactic symptoms, plasma histamine levels, and histology of intestines and lungs after oral challenge were examined.. Epicutaneous (EC) sensitization of mice to OVA induced a high level of OVA-specific IgE. Subsequent oral challenge with OVA resulted in symptoms of systemic anaphylaxis with elevated levels of plasma histamine as well as histological changes in both intestines and lungs. In the presence of anti-IL-4 antibodies, EC sensitization failed to provoke an IgE response, but still induced a Th2-predominant cellular immune response in lungs after oral challenge.. We demonstrated for the first time that food allergy can be induced by allergen exposure through the skin. Our results identify a novel role of EC sensitization in the pathogenesis of food allergy.

Topics: Administration, Cutaneous; Administration, Oral; Allergens; Anaphylaxis; Animals; Female; Food Hypersensitivity; Histamine; Immunoglobulin E; Interleukin-4; Intestines; Lung; Mice; Mice, Inbred BALB C; Ovalbumin; Th2 Cells

Recent studies have developed a murine model of IgE-mediated food allergy based on oral coadministration of antigen and cholera toxin (CT) to establish a maximal response for studying immunopathogenic mechanisms and immunotherapeutic strategies. However, for studying subtle immunomodulating factors or factors effective during response initiation, this maximal response-based model is less suitable due to a lack of sensitivity. Therefore, in attempts to identify essential parameters to fine-tune the immune response towards a submaximal level, potentially more sensitive, we were interested in characterizing the individual effects of the parameters in the CT-based model: CT dose, antigen type and dose, and number of immunizations.. BALB/c mice were orally sensitized weekly for 3 or 7 weeks with graded doses of CT and various food antigens (soy-trypsin inhibitor, ovalbumin or ovomucoid). Antigen-specific IgG1, IgG2a, IgA and IgE were monitored by ELISA.. The CT dose exerted a clear dose-dependent effect on the antigen-specific antibody response whereas the antigen dose tended to affect the kinetics of the developing response. Both the intensity and kinetics of the antibody response depended on the type of antigen and number of immunizations.. The critical parameters of the CT-based murine allergy model differentially control the intensity and kinetics of the developing immune response. Adjustment of these parameters could be a key tool for tailoring the response to submaximal levels rendering the model potentially more sensitive for evaluating the effect of subtle immunomodulating factors that would be lost in the maximal response-based model.

Topics: Animals; Cholera Toxin; Disease Models, Animal; Dose-Response Relationship, Immunologic; Enzyme-Linked Immunosorbent Assay; Female; Food Hypersensitivity; Immunization; Immunoglobulin E; Mice; Mice, Inbred BALB C; Ovalbumin; Ovomucin; Th2 Cells; Trypsin Inhibitor, Kunitz Soybean

Food allergy is a common disease without effective treatment. Since strict elimination of food allergens may be difficult, strategies for effective intervention are urgently needed.. The aim was to investigate the prophylactic use of orally administrated FIP-fve, an immunomodulatory protein isolated from the edible mushroom Flammulina velutipes, in a murine model of food allergy.. BALB/c mice were immunized twice intraperitoneally with ovalbumin (OVA), at an interval of 2 weeks. Before and during each period of immunization, FIP-fve (200 microg per mouse) or phosphate-buffered saline was given orally every other day with a total of five doses. Then OVA-specific antibodies and cytokine profiles were determined. Subsequently, the mice were orally challenged with OVA. Symptoms of anaphylaxis, levels of plasma histamine, and histology of intestines were examined.. Mice receiving oral FIP-fve treatment during sensitization to OVA had an impaired OVA-specific IgE response with a Th1-predominant cytokine profile. These mice were protected from systemic anaphylaxis-like symptoms induced by subsequent oral challenge with OVA.. Oral administration of FIP-fve has a Th1-skewing effect on the development of the allergen-specific immune response, and may serve the purpose of immunoprophylaxis for food allergy and other allergic diseases.

Topics: Administration, Oral; Anaphylaxis; Animals; Antigen-Presenting Cells; Cell Division; Female; Food Hypersensitivity; Fungal Proteins; Histamine; Immunoglobulin E; Interferon-gamma; Interleukin-4; Jejunum; Lectins; Lymphocyte Activation; Mice; Mice, Inbred BALB C; Ovalbumin; Th1 Cells

This study was carried out to evaluate the changes in the allergenic and antigenic properties of hen's egg albumin (ovalbumin [OVA]) with the combination of heat and gamma irradiation treatment. OVA solution samples were treated by (i) heating (sample 1), (ii) irradiation after heating (sample 2), and (iii) heating after irradiation (sample 3). Samples were isothermally heated and irradiated at the absorption dose of 10 kGy. Competitive indirect enzyme-linked immunosorbent assays (ELISAs) were performed with blood serum to test the ability of treated OVA to bind to immunoglobulin E (IgE) and mouse murine monoclonal antibody (IgG). OVA's ability to bind to mouse IgG changed upon heating at 75 degrees C, and its ability to bind to egg-allergic IgE changed upon heating at 80 degrees C. The ELISAs showed that egg-allergic IgE did not recognize OVA very well when heated at > or = 80 degrees C, while mouse IgG retained better activity under these conditions. Egg-allergic IgE binding was low both for OVA samples treated by heating and for samples treated by irradiation followed by heating. These results show that allergies induced by OVA could be effectively reduced by the combination of heat and gamma irradiation treatment.

Topics: Animals; Binding, Competitive; Egg Proteins; Enzyme-Linked Immunosorbent Assay; Food Hypersensitivity; Gamma Rays; Hot Temperature; Humans; Immunoglobulin E; Immunoglobulin G; Mice; Ovalbumin; Protein Denaturation

Information on the comparative digestibility of food allergens and nonallergenic proteins is crucial when stability to digestion is to be used as a criterion to assess the allergenic potential of novel proteins. In this work, we compared the digestive stability of a number of food allergens and proteins of unproven allergenicity and examined whether allergens possess a higher stability than nonallergenic proteins of similar cellular functions, and whether there is a correlation between protein digestibility and allergenicity. The stability of groups of storage proteins, plant lectins, contractile proteins, and enzymes, both allergens and proteins with unproven allergenicity, in a standard simulated gastric fluid and a standard simulated intestinal fluid was measured. Food allergens were not necessarily more resistant to digestion than nonallergenic proteins. There was not a clear relationship between digestibility measured in vitro and protein allergenicity.

Topics: Allergens; Body Fluids; Dietary Proteins; Digestion; Drug Stability; Electrophoresis, Polyacrylamide Gel; Food Hypersensitivity; Gastric Mucosa; Intestinal Mucosa; Lactoglobulins; Models, Biological; Ovalbumin; Papain; Pepsin A; Plant Lectins

The ability of food proteins to resist digestion in simulated gastric fluid (SGF) correlates with allergenic potential. The purpose of the current investigations was to determine whether this association is due solely to the failure of unstable proteins to elicit an immune response when administered orally. We have examined immune responses induced in BALB/c mice by gavage administration of ovalbumin (OVA) and a crude potato protein extract (PPE) containing acid phosphatase activity. The stability of OVA and PPE in SGF was measured using sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis. The ability of these proteins to stimulate specific IgG and IgE antibody production in mice following parenteral (intraperitoneal; ip) or oral (gavage) exposure was compared using enzyme-linked immunosorbent and homologous passive cutaneous anaphylaxis assays, respectively. Both OVA and PPE induced specific IgG antibody responses when administered either by gavage or by ip injection. Parenteral, but not gavage, exposure to OVA was associated with robust IgE antibody responses. Administration of PPE failed to stimulate strong IgE production via either route of exposure. Differential stability in SGF was observed, with PPE being digested extremely rapidly (within 1 min), whereas OVA was more resistant. The strong association reported by others between stability in SGF and allergenic potential is unlikely to be solely due to orally-ingested labile proteins failing to provoke immune responses due to degradation in the stomach.

Topics: Acid Phosphatase; Animals; Dietary Proteins; Digestion; Food Hypersensitivity; Gastric Lavage; Gastric Mucosa; Immunoglobulin E; Immunoglobulin G; Male; Mice; Mice, Inbred BALB C; Ovalbumin; Pepsin A; Solanum tuberosum; Stomach

Our previous study using allergen-sensitized murine splenocyte cultures has shown that Lactobacillus casei strain Shirota (LcS), a lactic acid bacterium widely used as a starter for fermented milk products, suppresses IgE production through promoting a dominant Th1-type response mediated by IL-12 induction.. We tried to evaluate the ability of LcS to suppress both IgE response and allergic reactions in vivo using a food allergy model with ovalbumin-specific T cell receptor transgenic (OVA-TCR-Tg) mice.. The ability of heat-killed LcS to induce IL-12 in serum was tested. OVA-TCR-Tg mice were fed a diet containing OVA for 4 weeks and injected with LcS intraperitoneally three times in the first week of this period. Cytokine and antibody secretion by splenocytes, and serum IgE and IgG1 responses were examined. The inhibitory effect of LcS on systemic anaphylaxis induced by intravenous challenge of OVA-fed OVA-TCR-Tg mice with OVA was also tested.. Intraperitoneal injection of LcS induced an IL-12 response in the serum of OVA-TCR-Tg mice. In the food allergy model, LcS administration skewed the pattern of cytokine production by splenocytes toward Th1 dominance, and suppressed IgE and IgG1 secretion by splenocytes. The ability of LcS to modulate cytokine production was blocked by anti-IL-12 antibody treatment. LcS also inhibited serum OVA-specific IgE and IgG1 responses and diminished systemic anaphylaxis.. LcS administration suppresses IgE and IgG1 responses and systemic allergic reactions in a food allergy model, suggesting a possible use of this lactic acid bacterium in preventing food allergy.

Topics: Anaphylaxis; Animals; Antibodies; Cells, Cultured; Cytokines; Food Hypersensitivity; Genes, T-Cell Receptor; Immunoglobulin E; Immunoglobulin G; Interleukin-12; Lacticaseibacillus casei; Male; Mice; Mice, Inbred BALB C; Mice, Transgenic; Ovalbumin; Peptide Fragments; Spleen; Th1 Cells

Topics: Adult; Double-Blind Method; Eosinophils; Female; Food Hypersensitivity; Histamine Release; Humans; Kinetics; Male; Mast Cells; Methylhistamines; Milk Hypersensitivity; Nuts; Ovalbumin; Proteins; Radioallergosorbent Test; Radioimmunoassay; Skin Tests

For the safety evaluation of genetically engineered crops the potential allergenicity of the newly introduced protein(s) has become an important issue. There is, however, no universal and reliable test system for the evaluation of the allergenic potency of food products. The best known allergy assessment proposal is the careful stepwise process using the IFBC/ILSI decision tree. Unfortunately, the described tests are not always conclusive, especially if the gene source coding for the protein has no history of dietary use and/or an unknown history in terms of allergenicity. The further testing warranted should in particular be focused on the prediction of the sensitizing potential of the novel protein, for which animal models are considered to be needed. In this paper the results are summarized of a promising food allergy model developed in Brown Norway (BN) rats. The results demonstrate that BN rats can be sensitized orally to the various allergenic food proteins tested, resulting in significant antigen-specific IgE responses, without the use of adjuvants. Upon oral challenge of previously sensitized animals, local and systemic immune-mediated effects, such as increased gastrointestinal permeability and decreased breathing frequency and blood pressure, could also be observed.

Topics: Animals; Antibody Formation; Disease Models, Animal; Egg Proteins; Food Hypersensitivity; Lactoglobulins; Male; Milk Proteins; Ovalbumin; Rats; Rats, Inbred BN

There is a need to identify and characterize the allergenic potential of novel proteins introduced into genetically-modified crop plants. Although several approaches have already been described, none of these measures directly the ability of proteins to cause allergic sensitization. For this reason there has been a growing interest in the development of suitable animal models. This article describes experience to date with a method based upon assessment of serological (IgG and IgE antibody) responses induced in BALB/c strain mice by proteins. Comparisons have been made between intraperitoneal (i.p.) administration and exposure by gavage using both allergenic and non-allergenic proteins. The available data indicate that responses provoked by i.p. exposure permit the identification of proteins that have the inherent potential to induce IgE antibody production and allergic sensitization. Moreover, this approach also provides a rank order of proteins with respect to allergenic potency that apparently reflects what is known of their relative sensitizing activity in humans. By comparison, oral exposure of mice by gavage is somewhat less sensitive. On this basis it is proposed that the inherent sensitizing potential of novel proteins can be evaluated as a function of IgE antibody responses stimulated by parenteral (i.p.) exposure of BALB/c mice.

Topics: Acid Phosphatase; Animals; Food Hypersensitivity; Immunoglobulin E; Mice; Mice, Inbred BALB C; Ovalbumin; Peanut Agglutinin; Plant Lectins; Solanum tuberosum

The trend towards an increased consumption of minimally processed plant food results in a higher intake of non-nutritive compounds such as lectins. Lectins are typically globular proteins that are resistant to digestion in the gastrointestinal tract. They affect the integrity of the intestinal epithelium and the absorption of dietary antigens, and induce the release of allergic mediators from mast cells in vitro. Based on this information we have studied whether dietary wheat germ agglutinin (WGA) could be involved in triggering food allergies. Brown Norway rats were immunized intraperitoneally using ovalbumin (OVA; 10 microg/rat) and 10 d later treated for five consecutive days with WGA (10 mg/rat per d) administered intragastrically. Rats were then orally challenged with OVA (100 microg/rat) 1 h after the last WGA application, and blood was collected 4 h later. Immunological responses (anti-OVA immunoglobulins E and G, rat mast cell protease II, interferon-gamma and lymphocyte proliferation) were measured and lymphocyte subpopulations were determined. In immunized rats WGA treatment resulted in increased serum rat mast cell protease II concentrations (pre-challenge 0.26 (SE 0.08) microg/ml, post-challenge 0.49 (SE 0.09) microg/ml; P < 0.01) 4 h after the OVA challenge. After 5 d serum concentrations of anti-OVA immunoglobulin E were significantly increased only in the immunized controls (absorbance at 405 nm on days 14 and 19 was 0.09 (SE 0.008) and 0.24 (SE 0.046) respectively; P = 0.02), while in WGA-treated rats no significant increase was seen (0.08 (SE 0.004) and 0.15 (SE 0.037 respectively; P = 0.14). CD4+ : CD8+ T lymphocytes in the spleen was significantly increased at this time (OVA 1.1 (SD 0.2), 1.4 (sd 0.1), P < 0.05). The treatment did not impair the proliferation and interferon-gamma production of mesenteric lymphocytes. In conclusion, these data suggest that high dietary intake of lectins such as WGA may affect the allergic response towards oral antigens in the gut-associated lymphoid tissue.

Topics: Animals; Caco-2 Cells; CD4-Positive T-Lymphocytes; CD8-Positive T-Lymphocytes; Cell Culture Techniques; Diet; Food Hypersensitivity; Humans; Immune Tolerance; Immunoglobulin E; Interferon-gamma; Lymphocyte Activation; Male; Mast Cells; Mesentery; Metalloendopeptidases; Ovalbumin; Rats; Rats, Inbred BN; Wheat Germ Agglutinins

A relationship between nonspecific bronchial hyperresponsiveness and allergic airway inflammation has been reported in children and in adults with asthma, but the relationship in infants with asthma is still unclear.. To evaluate the relationship between bronchial hyperresponsiveness and total serum IgE level throughout childhood. Bronchial reactivity to methacholine from the age of 1 to 16 years was studied by methacholine inhalation challenge using transcutaneous oxygen pressure (tcPO2) monitoring.. Two hundred one asthmatic children (boys:girls = 132:69; 7.3+/-4.0 years of age, mean +/- SD) were enrolled in this study. The tcPO2 was measured using a tcPO2 monitor. Serial doses of methacholine were doubled until a 10% decrease in tcPO2 from the baseline was reached. The cumulative dose of methacholine at the inflection point of tcPO2 was considered to represent the bronchial reactivity to methacholine.. There was no relationship between the cumulative dose of methacholine at the inflection point of tcPO2 and total serum IgE level in the group of children aged 1 to 4 years (P = 0.212), but significant correlations were found in the groups aged 5 to 10 years and 11 to 16 years (P = 0.044 and P = 0.014, respectively).. We conclude that there is an age-dependent relationship between bronchial reactivity to methacholine and the total serum IgE level and that inhaled allergens, which were more common allergens in older children, may have some effects on the degree of bronchial reactivity to methacholine in children with asthma.

Topics: Administration, Inhalation; Adolescent; Age Factors; Allergens; Animals; Asthma; Bronchial Hyperreactivity; Child; Child, Preschool; Female; Food Hypersensitivity; Humans; Immunoglobulin E; Infant; Male; Methacholine Chloride; Milk; Ovalbumin; Oxygen; Partial Pressure; Radioallergosorbent Test

This study was designed to evaluate the role of c-sensitive fibers in the establishment of immune-induced flavor aversion in mice.. Mice were treated neonatally with capsaicin in order to destroy c-sensitive fibers; after such treatment, adult animals, immunized or not with ovalbumin, were submitted to a two-bottle preference test, with a choice between water and a sweetened egg white solution.. Neonatal capsaicin treatment was unsuccessful in preventing the development of immune-induced aversion to the sweetened solution containing the antigen. Nonetheless, amongst immunized mice, those which had been previously treated with capsaicin showed a significant increment in the preference for the sweetened egg white solution. Furthermore, our data showed that neonatal capsaicin treatment did not interfere with either IgG1 or IgE production.. The present results suggest that c-sensitive fibers have a role in the transmission of the signals generated by this immune response to the central nervous system, thus contributing to the development of a flavor aversion in mice.

Topics: Animals; Animals, Newborn; Avoidance Learning; Capsaicin; Denervation; Food Hypersensitivity; Food Preferences; Immunoglobulin E; Immunoglobulin G; Male; Mice; Mice, Inbred BALB C; Nerve Fibers; Neuroimmunomodulation; Ovalbumin; Synaptic Transmission; Taste

There is a growing interest in the development of methods to characterize the allergenic properties of novel proteins, particularly those expressed by transgenic crop plants. Approaches to the direct evaluation of allergenic potential have focused generally on the ability of proteins to induce antibody (particularly IgE antibody) after systemic (intraperitoneal; i.p.) or gavage administration to high IgE responder strain rodents. To date there has been no systematic comparison of the reliability, sensitivity or selectivity of these approaches. We have, therefore, compared antibody (IgG and IgE) responses induced in Brown Norway (BN) rats by daily gavage administration and in BALB/c strain mice following intraperitoneal or gavage exposure to food proteins of varying allergenic potential. Animals were exposed to the allergens peanut agglutinin and ovalbumin (OVA) or to a crude potato protein extract (PPE) containing acid phosphatase activity, a common foodstuff which appears to be of low allergenicity. All test proteins were clearly immunogenic when administered by gavage to BN rats, with measurable, and in some cases very vigorous, IgG antibody responses recorded for all animals. Identical exposure of BALB/c strain mice also stimulated detectable IgG antibody responses, with particularly high titers recorded following treatment with peanut agglutinin and somewhat less vigorous responses induced by OVA and PPE. Despite these high titer IgG antibody responses, however, none of the proteins provoked detectable IgE antibody following gavage administration to BN rats. In contrast, in BALB/c mice oral exposure to peanut agglutinin elicited high titer IgE antibody, although IgE antibody responses to both OVA and PPE were much weaker. Parenteral (i.p.) treatment of BALB/c strain mice with each of the test materials induced relatively high titer IgG antibody and a differential potential to stimulate IgE antibody was observed. High titer IgE responses were provoked by i.p. administration of peanut agglutinin and OVA, whereas PPE stimulated little or no detectable IgE antibody. It would appear, therefore, that while it is possible to elicit robust IgE responses by gavage exposure of BALB/c strain mice to some protein allergens, such as peanut agglutinin, such responses are generally weaker and less consistent than those provoked by i.p. administration. Furthermore, gavage treatment failed to induce detectable IgE responses in the BN rat, suggesting that the ability t

Topics: Administration, Oral; Animals; Antibody Formation; Dietary Proteins; Female; Food Hypersensitivity; Immunoglobulin E; Immunoglobulin G; Injections, Intraperitoneal; Mice; Mice, Inbred BALB C; Ovalbumin; Peanut Agglutinin; Plant Lectins; Rats; Rats, Inbred BN; Solanum tuberosum; Species Specificity

The risks associated with IgE-mediated food allergy highlight the need for methods to screen for potential food allergens. Clinical and immunological tests are available for the diagnosis of food allergy to known food allergens, but this does not extend to the evaluation, or prediction of allergenicity in novel foods. This category, includes foods produced using novel processes genetically modified (GM) foods, and foods that might be used as alternatives to traditional foods. Through the collation and analysis of the protein sequences of known allergens and their epitopes, it is possible to identify related groups which correlate with observed clinical cross-reactivities. 3-D modelling extends the use of sequence data and can be used to display eptiopes on the surface of a molecule. Experimental models support sequence analysis and 3-D modelling. Observed cross-reactivities can be examined by Western blots prepared from native 2-D gels of a whole food preparation (e.g. hazelnut, peanut), and common proteins identified. IgEs to novel proteins can be raised in Brown Norway rat (a high IgE responder strain) and the proteins tested in simulated digest to determine epitope stability. Using the CSL serum bank, epitope binding can be examined through the ability of an allergen to cross-link the high affinity IgE receptor and thereby release mediators using in vitro cell-based models. This range of methods, in combination with data mining, provides a variety of screening options for testing the potential of a novel food to be allergenic, which does not involve prior exposure to the consumer.

Topics: Allergens; Animals; Cooking; Cross Reactions; Databases, Protein; Epitopes; Food; Food Hypersensitivity; Humans; Immunoblotting; Immunoglobulin E; Mast Cells; Nut Hypersensitivity; Ovalbumin; Peanut Hypersensitivity; Rats; Rats, Inbred BN

Factors that either protect from or enhance the development of atopic disease appear to be acting early in life. The gestational environment, including maternal immune responses, such as transplacentally transferred immunoglobulin (Ig) G antibodies to allergens, may be of importance in this respect, since allergen-specific immunity has been demonstrated to develop in utero.. To evaluate the relation between cord blood IgG subclass antibodies to allergens, maternal atopy and development of atopic disease in the children.. The study group comprised a cohort of 96 children participating in a prospective study up to 8 years of age. Cord blood IgG subclass antibodies to ovalbumin, beta-lactoglobulin, Bet v 1 and cat dander were analysed by ELISA.. The levels of all IgG subclass antibodies to ovalbumin and rBet v 1 were higher in newborn infants with an atopic mother, as compared with babies with nonatopic mothers. IgG1 antibody levels to cat and IgG4 antibody levels to beta-lactoglobulin and cat were also higher in atopic than in nonatopic mothers, whereas the other subclass antibody levels to those allergens were similar. High levels of cord blood IgG antibodies to cat and birch, but not to the food allergens, were associated with less atopic symptoms in the children during the first 8 years of life. Moreover, children who developed IgE antibodies to cat had lower levels of IgG antibodies to that allergen at birth.. High levels of cord blood IgG subclass, especially IgG4, antibodies to food and inhalant allergens are associated with maternal atopy. High levels of IgG antibodies to inhalant, but not food, allergens are associated with less development of atopy in the children.

Topics: Administration, Inhalation; Allergens; Animals; Antigens, Plant; Cats; Child; Female; Fetal Blood; Food Hypersensitivity; Humans; Hypersensitivity, Immediate; Immunoglobulin G; Immunoglobulin Isotypes; Infant; Infant, Newborn; Lactoglobulins; Ovalbumin; Plant Proteins; Pregnancy

No adequate enteral sensitization models are available to study food allergy and the allergenicity of food proteins. To further validate an enteral brown Norway (BN) rat sensitization model under development, we studied specific protein recognition to determine whether a comparable pattern of proteins is recognized by the rat immune system and the human immune system.. The animals were exposed to either ovalbumin as a positive reference control, hen's egg-white-protein extract, or a cow's milk preparation by daily gavage dosing (0.5, 1, 2.5, 5, 10, or 15 mg protein per rat/day) for 9 weeks. No adjuvants were used during the sensitization studies. The specificities of antibodies against hen's egg-white proteins or cow's-milk proteins in sera from orally sensitized rats and food-allergic patients were studied and compared by immunoblotting.. The IgG and IgE antibodies to hen's egg-white proteins and cow's-milk proteins present in sera from orally sensitized rats and food-allergic patients showed a comparable pattern of protein recognition.. Upon daily intragastric exposure to food allergens, the specificities of the induced antibody responses in the BN rat resemble those found in food-allergic patients. These studies add further support to the hypothesis that the BN rat may provide a suitable animal model for food allergy research and research on the allergenicity of food proteins.

Topics: Allergens; Animals; Antibody Specificity; Blotting, Western; Child; Child, Preschool; Dietary Proteins; Electrophoresis, Polyacrylamide Gel; Enzyme-Linked Immunosorbent Assay; Food Hypersensitivity; Humans; Immune System; Immunoglobulin E; Infant; Male; Milk; Milk Hypersensitivity; Milk Proteins; Ovalbumin; Passive Cutaneous Anaphylaxis; Rats; Rats, Inbred BN

The possible protective effect of breast milk against atopic manifestations in infancy, i.e. atopic eczema and food allergy, has been controversial for the last decades. Besides the methodological problems, differences in the composition of human milk could explain these controversies. The aim of this study was to investigate the composition of polyunsaturated fatty acids (PUFA) and secretory immunoglobulin A (S-IgA) levels to food proteins (ovalbumin and beta-lactoglobulin) and an inhalant allergen (cat) in milk from mothers of allergic and non-allergic children. Blood samples were obtained at birth and at 3 months from 120 children. Skin prick tests were performed at 6, 12 and 18 months, and the development of atopic diseases was assessed in the children. Breast milk samples were collected from their mothers at birth and monthly during the lactation period. Milk PUFA composition was measured by gas chromatography, and enzyme-linked immunosorbent assay (ELISA) was used to measure total S-IgA, anti-cat S-IgA, anti-ovalbumin S-IgA, and anti-beta-lactoglobulin S-IgA. Allergic disease developed in 44/120 children (22/63 children of allergic mothers and 22/57 children of non-allergic mothers). Lower levels of eicosapentaenoic acid, C20:5 n-3 (EPA), docosapentaenoic acid C22:5 n-3 (DPA), and docosatetraenoic acid C22:4 n-6 (DHA) (p < 0.05 for all) were found in mature milk from mothers of allergic as compared to milk from mothers of non-allergic children. The total n-6:total n-3 and the arachidonic acid, C20:4 n-6 (AA):EPA ratios were significantly lower in transitional and mature milk from mothers of allergic children, as compared to milk from mothers of non-allergic children. The PUFA levels in serum of allergic and non-allergic children were largely similar, except for higher levels of C22:4 n-6 and C22:5 n-6 (p < 0.05 for both) and a higher AA:EPA ratio in serum phospholipids in the former group (p < 0.05). Changes in the levels of milk PUFA were reflected in changes in PUFA serum phospholipids, particularly for the n-6 PUFA. The AA: EPA ratio in maternal milk was related, however, to the AA:EPA only in serum from non-allergic children, while this was not the case in allergic children. The levels of total S-IgA, anti-cat S-IgA, anti-ovalbumin S-IgA, and anti-beta-lactoglobulin S-IgA in milk from mothers of allergic, as compared to non-allergic, children were similar through the first 3 months of lactation. Low levels of n-3 PUFA in human milk, and particul

Topics: Animals; Breast Feeding; Cats; Child, Preschool; Fatty Acids, Unsaturated; Female; Food Hypersensitivity; Humans; Immunoglobulin A, Secretory; Infant; Infant, Newborn; Lactoglobulins; Milk, Human; Ovalbumin; Phospholipids

Systemically primed BALB/c mice developed severe diarrhea after repeated oral administration of ovalbumin (OVA). Histological analysis demonstrated that dramatic infiltration of eosinophils and mast cells occurred in the large intestine but not in the small intestine of mice with diarrhea. Interestingly, CD4(+) alphabeta T cells of the large intestine secreted IL-4 and IL-13 at high levels. Identically treated STAT6 gene-disrupted mice failed to develop OVA-induced diarrhea. Further, treatment of BALB/c mice with monoclonal anti-IL-4 antibody prevented the development of allergic diarrhea. An adoptive transfer study showed that systemically primed splenic CD4(+) T cells were preferentially recruited into the large intestine upon exposure to oral OVA. These results strongly suggest that systemically derived CD4(+) alphabeta T cells of the large intestine play a critical role in the onset of Th2-mediated intestinal allergic disorders via STAT6 signal transduction.

Topics: Adoptive Transfer; Animals; B-Lymphocytes; Diarrhea; Eosinophils; Food Hypersensitivity; Interleukin-13; Interleukin-4; Intestine, Large; Mast Cells; Mice; Mice, Inbred Strains; Mice, SCID; Ovalbumin; Receptors, Antigen, T-Cell, alpha-beta; Signal Transduction; Spleen; STAT6 Transcription Factor; T-Lymphocyte Subsets; Th2 Cells; Trans-Activators

Mast-cell-deficient W/W(v) mice were sensitized by oral administration of 0.1 and 1.0 mg ovalbumin (OVA) by gavage every day for 9 weeks, and active systemic anaphylaxis (ASA) was induced by intraperitoneal injection of OVA. The production of OVA-specific IgE and IgG1 by oral immunization of the W/W(v) mice was high, and the production of IL-4 by splenocytes re-stimulated with OVA in vitro was increased. In contrast, production of OVA-specific IgG2a and IgG2b was low, and production of IFN-gamma by splenocytes after re-stimulation with OVA in vitro was rather decreased. These findings suggest that Th2-dominant helper T-cell activation had occurred. No increase in serum histamine level was observed following ASA induction. However, the plasma platelet-activating factor (PAF) levels of the mice sensitized with 0.1 and 1.0 mg OVA by gavage increased significantly. The increases in plasma PAF correlated well with the ASA-associated decreases in body temperature, suggesting that PAF plays an important role in ASA in W/W(v) mice. Taken together the above findings indicate that W/W(v) mice are a good model not only for studying induction of food allergy but also for examining the role of PAF in food-induced hypersensitivity.

Topics: Administration, Oral; Anaphylaxis; Animals; Body Weight; Disease Models, Animal; Female; Fever; Food Hypersensitivity; Histamine; Injections, Intraperitoneal; Interferon-gamma; Interleukin-4; Mast Cells; Mice; Mice, Mutant Strains; Organ Size; Ovalbumin; Platelet Activating Factor; Th2 Cells

For many years the central focus of research into gastrointestinal hypersensitivity reactions has been the mast cell population of the intestinal lamina propria. Since bile is known to deliver immunological mediators to the gastrointestinal tract, the possibility arises that extra-intestinal populations of mast cells may also contribute to IgE-mediated intestinal damage.. To characterize hepatic mast cells in the rat and to investigate the role of the hepatobiliary system in a model of IgE-mediated reactivity to dietary antigen.. Wistar rats were passively sensitized with monoclonal antidinitrophenyl (DNP) IgE antibodies, and were later challenged orogastrically with DNP-HSA. Additional animals were sensitized, then bile duct-cannulated prior to antigen challenge. At various time points, liver and intestinal samples were collected for histological examination, and bile was collected and assayed for histamine and TNFalpha.. Hepatic mast cells display a mucosal mast cell-like phenotype, and are closely associated with the vessels of the portal triads. Orogastric antigen challenge led to a rapid and significant decline (P<0.0001) in detectable mast cells as a result of anaphylactic degranulation. The median number of granulated mast cells associated with each portal triad in liver sections declined from six per portal triad to one per portal triad post-antigen challenge. After 15 min, biliary histamine concentrations rose above background levels (P<0.01). TNFalpha was also detectable in the majority (4/6) of bile samples within 15 min of challenge. Histological examination of the gastrointestinal mucosa revealed disruption to the villous epithelium ranging from oedematous changes to gross destruction. Such damage was not seen in animals in which bile had been externally drained.. The data indicate that biliary products are major contributors to the gastrointestinal damage arising from IgE-mediated hypersensitivity reactions in the rat, and such hypersensitivity reactions may involve a population of mast cells which reside in the liver.

Topics: Animals; Bile; Biliary Tract; Blotting, Western; Cell Count; Dinitrophenols; Enteritis; Food Hypersensitivity; Histamine; Immunoglobulin E; Liver; Mast Cells; Ovalbumin; Radioimmunoassay; Rats; Rats, Wistar; Serum Albumin; Tumor Necrosis Factor-alpha

Although several in vivo antigenicity assays using parenteral immunization are operational, no full validated enteral models are available to study food allergy and allergenicity of food proteins. To further validate a developed enteral Brown Norway (BN) rat food allergy model, systemic and local immune-mediated reactions were studied upon oral challenges. The animals were exposed to ovalbumin (OVA) by daily gavage dosing (1 mg OVA/rat/day) for 6 weeks, without the use of an adjuvant, or by intraperitoneal injections with OVA together with AL(OH)3. Subsequently, effects on breathing frequency, blood pressure, and gastrointestinal permeability were investigated upon an oral challenge with 10 to 100 mg OVA in vivo. In both parenterally and orally sensitized rats, an increase in gut permeability (increased passage of beta-lactoglobulin as bystander protein) was determined between 0.5 and 1 h after an oral OVA challenge was given. An oral challenge with OVA did not induce a clear effect on the respiratory system or blood pressure in the majority of the animals. However, some animals demonstrated a temporary decrease in breathing frequency or systolic blood pressure. Upon oral challenge with OVA of orally and parenterally sensitized animals, local effects were observed in all animals whereas systemic effects were observed at a low frequency, which reflects the situation in food allergic patients after an oral challenge. These studies show that the BN rat provides a suitable animal model to study oral sensitization to food proteins as well as immune-mediated effects after oral challenge with food proteins.

Topics: Animals; Blood Pressure; Disease Models, Animal; Enzyme-Linked Immunosorbent Assay; Food Hypersensitivity; Immunoglobulin E; Intestinal Absorption; Male; Ovalbumin; Rats; Rats, Inbred BN; Respiratory Mechanics

Reports of allergy to bird meats are uncommon, and most have been in patients with "bird-egg syndrome.". We sought to evaluate 3 patients who reported allergic reactions to several avian meats, but who denied allergic reactions to eating eggs. The patients required yellow fever vaccine for entry into the military.. Patients were skin tested with commercial extracts of chicken, turkey, and egg, as well as with crude extracts made from dove and quail meat, and with yellow fever vaccine. Immunoblots for IgE antibody were performed by using the same materials used for skin testing plus extracts of duck and goose meat.. Skin tests were positive in all 3 patients to chicken, turkey, dove, quail, and yellow fever vaccine and negative to egg. This included some positive skin test responses to bird meats the patients denied ever having eaten. The vaccine was administered in graded doses. Immunoblots revealed IgE binding to several proteins of similar molecular weights in all of the avian meats but not to egg or yellow fever vaccine. Again, this included IgE antibody to some bird meats the patients denied ever having eaten.. Patients allergic to one bird meat may be allergic to others, including game birds, probably because of cross-reacting allergens. Such patients may have to exercise caution even when eating bird meats they have not previously ingested. The relationship of this allergy to yellow fever vaccine, if any, remains to be determined.

Topics: Adolescent; Adult; Allergens; Animals; Antibodies, Anti-Idiotypic; Blood Protein Electrophoresis; Cross Reactions; Erythema; Food Hypersensitivity; Geese; Humans; Hypersensitivity, Delayed; Immunoblotting; Immunoglobulin E; Male; Meat; Ovalbumin; Poultry; Quail; Skin Tests; Turkeys; Viral Vaccines; Yellow fever virus

A number of studies demonstrating the important role of interleukin-5 (IL-5) in eosinophil infiltration were reported. Antigen-induced eosinophil infiltrations to the trachea and skin were inhibited by pretreatment with monoclonal anti-IL-5 antibody. In this study, the role of IL-5 in eosinophil infiltration to the gut by oral challenge in mice is investigated. A marked eosinophil infiltration to the lamina propria was induced by oral challenge with ovalubumin (OVA) in Balb/c mice intraperitoneally sensitized with OVA, and peaked at 6 h after the oral challenge. Intraperitoneal preadministration of monoclonal anti-IL-5 antibody significantly decreased the eosinophil infiltration to the lamina propria. Furthermore, analysis by reverse transcription polymerase chain reaction (RT-PCR) demonstrated that IL-5 mRNA expression was induced in the lamina propria in an antigen-specific manner and the expression peaked at 6 h and declined thereafter. In-situ hybridization (ISH) revealed the presence of IL-5 mRNA positive cells at lesion site. As in bronchial mucosa and skin, IL-5 may play an important role in eosinophil recruitment to the lesion site in IgE mediated gut late phase reaction.

Topics: Animals; Antibodies, Monoclonal; Cell Movement; Chemotactic Factors, Eosinophil; Edema; Eosinophils; Female; Food Hypersensitivity; In Situ Hybridization; Interleukin-5; Intestinal Mucosa; Male; Mice; Mice, Inbred BALB C; Ovalbumin; Rats; Rats, Sprague-Dawley; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger

Food contaminants may contribute to the recent increased incidence of food allergies. We have investigated this hypothesis experimentally. It was our objective to determine whether toxicity to the intestinal tissue by orally applied mercury (Hg) could modulate the immune response to food allergens. Effective mechanisms were studied with functional immunological and toxicological parameters. Brown Norway rats were immunized intraperitoneally by ovalbumin (OVA). Before oral challenge with OVA, immunized and non-immunized animals were exposed to HgCl2. Immunological responses were measured by enzyme-linked immunosorbent assays [anti-OVA-IgE and-IgG, rat mast cell protease II (RMCPII), interferon-gamma, interleukin-4, lymphocyte proliferation] and by flow cytometry (lymphocyte subpopulations). Toxicity of Hg to the intestinal barrier was determined by measuring viability, DNA damage and induction of glutathione S-transferase in isolated intestinal epithelial cells and lymph node cells, and by measuring permeability, short-circuit current and tissue conductance of the intact intestinal epithelium. A single high oral dose of HgCl2 enhanced the serum concentrations of anti-OVA-IgE and IgG (P < 0.05) and of RMCPII (P < 0.05) in immunized rats. The treatment resulted in a higher number of CD4/CD25+ T cells in the lymph nodes (P < 0.05). The multiple application of low HgCl2 doses (5 x 0.2 mg/kg body weight) only resulted in an elevated RMCPII serum concentration (P < 0.05). Neither treatment schedules impaired proliferation and cytokine production of lymphocytes. In non-immunized rats only minor immunological changes were observed. Oral HgCl2 induced genotoxic damage in lymph node cells and in jejunal epithelial cells (P < 0.05). Moreover, HgCl2 increased the permeability of intestinal epithelial tissue and of Caco-2 monolayers and was genotoxic and cytotoxic to isolated intestinal epithelial cells in vitro. In conclusion, these studies indicate that the food contaminant Hg can stimulate the immune response to OVA in immunized rats. One possible mechanism could be the toxicity of Hg to the intestinal epithelial and the lymph node cells. Whether humans with allergies respond to high oral doses of Hg in a similar way needs to be investigated in further studies.

Topics: Animals; Antibody Formation; Antigens, Surface; Caco-2 Cells; Cell Membrane Permeability; DNA Damage; Enzyme Induction; Epithelial Cells; Fluorescein; Food Hypersensitivity; Glutathione Transferase; Humans; Immunity, Mucosal; Interferon-gamma; Interleukin-4; Intestinal Mucosa; Lymphocyte Activation; Male; Mast Cells; Mercury; Mesentery; Ovalbumin; Rats; Rats, Sprague-Dawley; Serine Endopeptidases

We developed an oral sensitization protocol for food proteins for the rat. Young Brown Norway (BN) rats were exposed to 1 mg ovalbumin (OVA) by daily gavage dosing for 42 days without the use of an adjuvant. OVA-specific IgE and IgG responses were determined by ELISA. On an oral challenge with OVA some clinical symptoms of food allergy-like effects on the respiratory system, blood pressure, and permeability of the gastrointestinal barrier were studied. In addition, BN rats were orally exposed to a total hen egg white protein (HEW) extract and cow's milk (CM) and the specificities of induced antibody responses were compared with the specificities of antibodies in sera from egg- and milk-allergic patients using immunoblotting. Animals orally exposed to the allergens developed specific IgE and IgG antibodies which recognized the same proteins compared with antibodies from egg- or CM-allergic patients. Among the various clinical symptoms of food allergy, gut permeability was increased after an oral challenge. In addition, some animals demonstrated a temporary decrease in breathing frequency or systolic blood pressure. The results obtained show that the Brown Norway rat is a suitable animal model for inducing specific IgG and IgE responses on daily intragastric dosing of OVA without the use of an adjuvant. Moreover, local immune-mediated effects on oral challenge are observed. The observation that enterally exposed BN rats and food-allergic patients demonstrate antibody responses to a comparable selection of proteins on exposure to different protein mixtures (HEW and CM) further supports the suitability of the BN rat as an animal model for food allergy research and for the study of the allergenicity of (novel) food proteins.

Topics: Administration, Oral; Allergens; Animals; Antibodies; Blood Pressure; Cattle; Chickens; Dietary Proteins; Digestive System; Disease Models, Animal; Egg Proteins; Food Hypersensitivity; Humans; Immunization; Immunologic Techniques; Male; Milk Proteins; Ovalbumin; Passive Cutaneous Anaphylaxis; Rats; Rats, Inbred BN; Respiratory Function Tests

A model of antigen-specific T-cell proliferative responses based on reciprocal patterns of responses to dietary and inhalant allergens has been suggested, the former being frequent in infancy but rare in adults, whereas the latter are preserved and expand between infancy and adulthood. We have evaluated the age-related variations of mononuclear cell reactivity to food allergens. The cord blood mononuclear cells (CBMC) of 30 neonates without family history of atopy and the peripheral blood mononuclear cells (PBMC) of 20 healthy children and of 40 healthy adults were stimulated in vitro with beta-lactoglobulin (BLG) or ovalbumin (OVA) and the cultures were harvested after 7 days. Neonates, children and adults were compared for the percentages of positive responses and for the magnitude of response. Adult subjects showed significantly lower percentages of positive responses and reduced magnitude of response than those observed in neonates and children either in BLG or in OVA cultures. We have not observed a decrease of food allergen mononuclear cell reactivity between neonates and children for the frequency of positive responses. The magnitude of response of neonates was significantly lower than that of children in BLG cultures. Our results seem to confirm the loss of mononuclear cell reactivity to food allergens in adult age. However, other reports show conflicting data. We suggest that a rigorous standardization of the methodological steps of in vitro mononuclear cell stimulation with allergen is necessary.

Topics: Adult; Aging; Allergens; Child, Preschool; Female; Fetal Blood; Food Hypersensitivity; Humans; In Vitro Techniques; Infant; Infant, Newborn; Lactoglobulins; Leukocytes, Mononuclear; Lymphocyte Activation; Male; Models, Biological; Ovalbumin; T-Lymphocytes

Although several in vivo antigenicity assays using parenteral immunization are operational, no adequate enteral sensitization models are available to study food allergy and allergenicity of food proteins.. This paper describes the development of an enteral model for food allergy research in the Brown Norway (BN) rat.. The animals were exposed to ovalbumin either ad libitum via the drinking water (0.002 to 20 mg/mL) continuously for 6 weeks or by gavage (1 mg/mL per rat). Gavage dosing was performed either daily, twice a week, once a week or once every 2 weeks during a period of 6 weeks. No adjuvants were used during the sensitization studies.. After intra-gastric administration of ovalbumin once or twice a week or once every two weeks, no or only a very low frequency of ovalbumin-specific antibody responses were detected. Daily intra-gastric dosing with ovalbumin resulted in antigen-specific IgG as well as IgE responses in almost all animals tested. Upon ad libitum exposure, ovalbumin-specific IgG but no ovalbumin-specific IgE was detected. The cellular response was examined by determination of delayed-type hypersensitivity (DTH) reactions in the animals dosed by daily gavage and in the ad libitum exposed rats. Both sensitization protocols sensitized for DTH. The response was most pronounced in ad libitum exposed rats at day 28 of exposure.. These studies show that the BN rat may provide a suitable animal model for inducing specific IgG and IgE responses as well as specific T-cell mediated hypersensitivity (DTH) to ovalbumin upon exposure via the enteral route without the use of adjuvants.

Topics: Administration, Oral; Animals; Disease Models, Animal; Enzyme-Linked Immunosorbent Assay; Food Hypersensitivity; Hypersensitivity, Delayed; Immunoglobulin E; Immunoglobulin G; Male; Ovalbumin; Passive Cutaneous Anaphylaxis; Rats; Rats, Inbred BN

Rats were sensitized with ovalbumin (OVA) with a molecular weight of 45 kd, challenged with OVA orally, followed by orally administered beta-lactoglobulin (BLG) as an intestinal permeability marker. BLG is a macro-molecular protein with a molecular weight of 18 kd. Blood BLG concentrations were measured (by ELISA) serially over 4 hours following BLG administration, which in turn was given 1 hour after OVA challenges. The maximum BLG concentration was at 2 hours. BLG was then administered orally 1, 3, 6, 12 and 24 hours after oral OVA challenge, and the serum BLG concentration at 2 hours after BLG administration was compared among the five groups. BLG appeared in the circulation of the animals 1, 6 and 24 hours after allergen challenge, but not after 3 and 12 hours. The serum BLG concentration was not significantly different at 1, 6 and 24 hours. Histopathological examinations of the intestines showed mast cell infiltration of the intestinal mucosa at 1 hour, remarkable edema of villi at 3 hours, eosinophil infiltration at 6 hours, an increase of goblet cells at 12 hours and villous atrophy and lymphocyte infiltration at 24 hours. The appearance in the serum of three BLG peaks of comparable heights suggested that the intestinal absorption of BLG may be related to a late and delayed phase as well as the immediate IgE-dependent phase response.

Topics: Administration, Oral; Animals; Food Hypersensitivity; Immunoglobulin E; Intestinal Absorption; Intestinal Mucosa; Intestines; Lactoglobulins; Male; Ovalbumin; Passive Cutaneous Anaphylaxis; Permeability; Rats; Rats, Inbred BN; Time Factors

Substantial part of patients who suffer from food allergy outgrow their allergic reaction. Moreover the mechanisms of this phenomenon are poorly understood. We studied cytokine mRNA expression in peripheral blood mononuclear cells (PBMC) from children with egg allergy, nine patients on active stage and eight were outgrown, and four healthy controls, by use of reverse transcription polymerase chain reaction. Following ovalbumin (OVA) stimulation in vitro, active patients demonstrated increasing IL-5 mRNA. In comparison, no increasing expression of IL-5 mRNA was observed in outgrown and healthy children. IL-4 and IFN-gamma mRNA expression has no tendency either to increase or to decrease in all three groups. There was no difference of proliferative responses for OVA among these groups suggesting that outgrown patients' PBMC did not fall into anergy or clonal deletion. These data suggested the change in balance of cytokine production of PBMC which were stimulated by allergen is a trigger for "outgrow" of food allergy.

Topics: Allergens; Child; Child, Preschool; Cytokines; Food Hypersensitivity; Humans; Infant; Leukocytes, Mononuclear; Ovalbumin; RNA, Messenger

We characterize the intraepithelial T cell population of terminal ileum from intraperitoneally OVA-sensitized New Zealand rabbit after oral challenge. High anti-OVA specific IgE levels elicited after sensitization were evaluated by positive passive cutaneous anaphylaxis (PCA) at 160 fold dilutions. Anaphylactic response in gut after the oral challenge was confirmed by evaluation of edema in villi and marked mast cell degranulatin. Total T cells (CD5+), T cells subset (CD4+) and MHC II R-DQ positive cells expressed as the number of positive cells/1000 nuclei, were analyzed for each tissue. Positive cells were counted in 30 villi by light microscopy. The number of CD5+ cells was 122.5 cells/1000 nuclei. Activated cells in small bowel epithelium 8 hours after challenge in experimental group, expressed by MHC II R-DQ showed an increase: 32.2 as compared to control group: 12.6 positive cells/1000 nuclei (p < 0.05) and compared to experimental group 1 hour after challenge 9.3 (p < 0.05). We demonstrated an increment of activated T cells in gut epithelium of terminal ileum in OVA-sensitized group after oral challenge.

Topics: Animals; Antibodies, Monoclonal; Food Hypersensitivity; Ileum; Immunoglobulin E; Intestinal Mucosa; Lymphocyte Count; Ovalbumin; Rabbits; Serine Proteinase Inhibitors; T-Lymphocytes

The antigen-specific interleukin-2 response (AIR) test using lymphocytes is effective in searching for the antigen which causes allergic diseases and understanding their disease activity.. The correlation between the raw egg oral provocation test and egg white antigen-specific interleukin-2 (IL-2) response test was investigated in 123 children with infantile atopic dermatitis and 13 children with bronchial asthma.. Among the 83 who showed positive reactions to provocation, 75 also reacted positively to the AIR test (sensitivity, 90.4%), while among the 53 children who showed negative responses to antigen provocation, 45 produced negative responses to the AIR test (specificity, 84.9%). The specificity of egg white IgE RAST score and skin-prick test are 88.7 and 81.3% which are comparable to that of the AIR test. However, their sensitivity was low (38.6 and 66.7%). In the patterns of symptom developed in the provocation AIR displayed late and delayed type allergic responses in addition to the immediate type which RAST reflected. The RAST-negative group composed of 98 patients included 51 (52.0%) who exhibited positive reactions to the provocation test. Among these 44 responded positively to the AIR test (86.3%).. The AIR test is effective for screening egg white antigen as part of the tests for antigens responsible for allergic diseases and as a test to ascertain the relevant antigens, and that the conditions that could not be diagnosed by RAST can be detected by the AIR test.

Topics: Administration, Oral; Adolescent; Antigens; Asthma; Child; Child, Preschool; Dermatitis, Atopic; Female; Food Hypersensitivity; Humans; Immunoglobulin E; Infant; Interleukin-2; Lymphocyte Activation; Male; Ovalbumin; Radioallergosorbent Test; Skin Tests; T-Lymphocytes

Topics: Allergens; Animals; Disease Models, Animal; Food Hypersensitivity; Immunoglobulin E; Milk Proteins; Ovalbumin; Rats; Rats, Inbred BN; Rats, Sprague-Dawley

Allergic reactions to food are characterized by enhanced allergen-specific IgE serum levels and the activation of intestinal mast cells. Here we describe a murine model for the onset of food allergy and the role of cytokines in the regulation of food-induced IgE responses.. Mice were primed systemically with low doses of alum-precipitated ovalbumin. Subsequent intragastric challenge led to enhanced sensitization.. Compared with baseline ovalbumin-specific IgE levels before challenge (0.23 +/- 0.06 optical density [OD] units), ovalbumin-challenged mice showed significantly elevated IgE levels (0.86 +/- 0.23 OD units) after intragastric challenge, which were not observed in control animals (0.29 +/- 0.06 OD units). IgE levels mirrored intestinal mast cell activation, measured by decreased histamine levels in duodenal specimens, in ovalbumin-challenged mice (92.6 +/- 7.9 ng/0.1 gm tissue weight) but not in saline-challenged mice (135.4 +/- 18.3 ng/0.1 gm tissue weight), compared with baseline levels (141.1 +/- 4.1 ng/0.1 gm tissue weight). Changes in IgE and histamine levels after intragastric challenge could be blocked by treating the animals with neutralizing antibodies against IL-4 or IL-10. Although it is generally accepted that ingestion of food allergens leads to a state of immunologic unresponsiveness (i.e., oral tolerance), it is shown here that low-dose systemic priming followed by intragastric challenge leads to sensitization instead of unresponsiveness.. Our murine model shows an important correlation between TH2 cytokines, IgE production, and histamine release. Hence, this in vivo model provides a useful tool with which the complex mechanism underlying sensitization to food allergens can be studied.

Topics: Allergens; Alum Compounds; Animals; Antibody Specificity; Cytokines; Disease Models, Animal; Female; Food Hypersensitivity; Histamine Release; Immunization; Immunoglobulin E; Interleukin-10; Interleukin-4; Intestine, Small; Mast Cells; Mice; Mice, Inbred BALB C; Neutralization Tests; Ovalbumin

To investigate the humoral immune response to common food antigens in Japanese children with IDDM.. IgG antibodies to cow's milk, beta-lactoglobulin, bovine serum albumin (BSA), alpha-lactalbumin, and hens egg ovalbumin were examined by enzyme-linked immunosorbent assay in the sera of 33 patients with IDDM, ages 11.8 +/- 3.4 years. The data were compared with that of 50 normal subjects, ages 10.3 +/- 5.1 years, who acted as control subjects. A positive antibody to a food antigen was defined as an antibody titer greater than the 95th percentile value in normal subjects.. Children with IDDM had significantly higher median titers of IgG antibodies to beta-lactoglobulin and ovalbumin (P = 0.03 and P = 0.0005 respectively). More children with IDDM than control subjects had positive IgG antibody to ovalbumin (21 vs. 6%, P = 0.04). Titers, as well as the number of positive antibodies to other food antigens, including BSA, did not differ between the two groups.. Japanese children with IDDM show an enhanced humoral immune response to beta-lactoglobulin and ovalbumin, a phenomenon that may be related to the pathogenesis of the disease.

Topics: Adolescent; Animals; Child; Diabetes Mellitus, Type 1; Food Hypersensitivity; Humans; Immunoglobulin G; Japan; Lactalbumin; Lactoglobulins; Milk; Ovalbumin; Serum Albumin, Bovine

We have attempted a new approach, using peripheral blood mononuclear cells (PBMCs), to predict the clinical course of infantile food-sensitive atopic dermatitis (AD). In this study, we investigated the relationships between the clinical course of infantile hen's-egg-sensitive AD and laboratory data obtained at the early stage of AD, particularly on combinations of specific IgE antibodies to hen's egg and proliferative responses of PBMCs to ovalbumin (OA). Total IgE concentrations, specific IgE antibodies to hen's egg and proliferative responses of PBMCs to OA were measured in 31 hen's-egg-sensitive AD patients within 6 months after development of AD symptoms. After the acquisition of laboratory data, the clinical courses of all patients were followed up for 1 year. The stimulation index of proliferative responses of PBMCs to OA in hen's-egg-sensitive AD patients whose symptoms did not improve was significantly (p < 0.05) higher than that in hen's-egg-sensitive AD patients whose symptoms improved. On the other hand, the radioallergosorbent test (RAST) scores for hen's egg were not statistically different between the two groups. The degree of refractoriness of AD symptoms tended to be higher in the patients who showed a higher total IgE concentration at the beginning of the clinical course than in those who showed a lower one. These results might be attributed to the underlying cause of AD, i.e., a cell-mediated and an IgE-mediated allergy. We concluded that, in addition to total IgE concentrations, combinations of RAST values and lymphocyte proliferative responses to OA are important to predict the clinical course of infantile hen's-egg-sensitive AD.

Topics: Allergens; Animals; Chickens; Child, Preschool; Dermatitis, Atopic; Eggs; Food Hypersensitivity; Humans; Immunoglobulin E; In Vitro Techniques; Infant; Lymphocyte Activation; Ovalbumin; Time Factors

The role of food-specific antibodies in the pathogenesis of food allergy is controversial. The first step in solving this controversy may be the assessment of antibody response to food antigens in the normal population. Most of the existing data in this field come from studies that used assays of different standards. This study investigated food-specific antibodies in the normal population using standardized assays. Normal levels of antibody titers were also derived for use as reference. Two hundred and eight individuals from different age groups participated. Immunoglobulin G (IgG) antibodies to cow's milk and its component proteins, and to hen's egg ovalbumin, IgA and IgM antibodies to beta-lactoglobulin and ovalbumin were measured by enzyme-linked immunosorbent assay. The sepharose-radioallergosorbent test was used to measure IgE antibodies to cow's milk and ovalbumin. Titers of IgG antibodies to cow's milk and its component proteins revealed an age-related trend, peaking in the 5 months-1 year age group and then decreased to negligible values in adults. A similar trend was observed with IgG anti-ovalbumin antibodies. Temporal association was less evident for antibodies of other classes. Only six subjects had positive IgE antibodies to cow's milk, while none had positive IgE anti-ovalbumin antibody. The prevalences of IgG antibodies to cow's milk, its component proteins, and ovalbumin are influenced by age and feeding habits. Cross-reactivity to related food antigens is common. The presence of IgE antibodies to food antigens is not a physiological phenomenon.

Topics: Animals; Antibodies; Child; Child, Preschool; Enzyme-Linked Immunosorbent Assay; Female; Food Hypersensitivity; Humans; Immunoglobulin A; Immunoglobulin E; Immunoglobulin G; Immunoglobulin M; Immunoglobulins; Infant; Infant, Newborn; Lactoglobulins; Male; Milk; Milk Hypersensitivity; Ovalbumin; Radioallergosorbent Test

The Brown Norway (BN) rat was examined as a model for investigating factors that influence the development of food allergy. An antigen dose-response curve for the production of antigen-specific reaginic antibody (IgE) induced through the oral route was determined. Animals were dosed orally with 1.0, 2.5, 5.0, 7.5, 10.0 and 12.0 mg ovalbumin/ml (0.5 ml/100 g twice a week for 6 wk). To promote IgE production the adjuvant carrageenan was administered once a week by the i.p. route. The effect on oral sensitization of 1.5 mg Gypsophila sp. saponin/ml administered together with the antigen on oral sensitization was examined in animals treated with 2.5, 6.0 or 10.0 mg ovalbumin/ml. The number of animals producing antigen specific reaginic antibody in response to 2.5 mg ovalbumin/ml was significantly increased (P < 0.01) in the group that received saponin with 2.5 mg ovalbumin/ml. These studies indicate that the BN rat is a sensitive model for the investigation of allergic reactions to food and has the potential to determine the impact of other dietary factors on the development of oral sensitization.

Topics: Adjuvants, Immunologic; Administration, Oral; Animals; Carrageenan; Disease Models, Animal; Dose-Response Relationship, Immunologic; Extravasation of Diagnostic and Therapeutic Materials; Food Hypersensitivity; Immunity, Mucosal; Immunoglobulin E; Immunoglobulin G; Injections, Intraperitoneal; Intestinal Mucosa; Intestines; Magnoliopsida; Male; Ovalbumin; Permeability; Plant Extracts; Rats; Rats, Inbred BN; Rats, Sprague-Dawley; Saponins

We studied allergenic determinants that induce hypersensitivity to OVA, the major allergen in egg allergy, using immunoblot and histamine release assays. Immunoblot analysis demonstrated a part of the OVA epitope was in the C-terminal region comprising residues 347-385 (OVA347-385). Histamine was released from basophils of a patient with egg allergy upon stimulation with the OVA fragment corresponding to OVA347-385. Furthermore, detailed epitope mapping using overlapping peptides (residues 347-366, OVA-A; residues 357-376, OVA-B; and residues 367-385, OVA-C) in the OVA 347-385 region was carried out using the histamine release assay. In order for histamine release from basophils to occur, the allergen must possess two or more allergenic determinants located on the protein molecule at distances that would be equivalent to the distances between IgE molecules on the membrane surface. these results suggest that there are at least two epitopes that bind IgE antibodies on each OVA peptide. In addition, one epitope that binds IgE antibodies in two patients appears to reside in the haptenic peptide OVA357-366 (OVA-B1). The histamine release from basophils stimulated by OVA-B was completely inhibited by OVA-B1 in one of these patients. Similarly, OVA-B1 inhibited the histamine release produced by OVA-A in the other by more than 40%. These results suggest that haptenic synthetic peptides could regulate the allergic reaction in the effector phase if common epitope(s) recognized by IgE antibodies in the patients with egg allergy can be found. These are the first studies that provide an antigen-specific approach to inhibiting histamine release from basophils by a haptenic peptide recognized by IgE antibodies in an allergic disorder.

Topics: Allergens; Amino Acid Sequence; Animals; Basophils; Chickens; Child; Child, Preschool; Eggs; Enzyme-Linked Immunosorbent Assay; Epitopes; Female; Food Hypersensitivity; Haptens; Histamine Release; Humans; Immunoblotting; Immunoglobulin E; Infant; Male; Molecular Sequence Data; Ovalbumin; Peptide Fragments; Serine Endopeptidases

In order to investigate T cell recognition of allergens in hen egg allergy, we have established 30 ovalbumin (OVA)-specific T cell lines (TCLs) from peripheral blood mononuclear cells of 6 patients with atopic dermatitis, who are positive for IgE antibodies to OVA and clinically allergic to hen egg, and characterized them for their cytokine production pattern. All TCLs we could study were mainly composed of CD4+ T cells. Most TCLs produced significant amounts of interleukin 4 (IL-4) and IL-5 but no or very little interferon gamma on antigen stimulation, suggesting that these TCLs belong to TH2-type T cells. Restriction elements and epitope specificities were further studied on some TCLs. Antibody blocking of the proliferative responses of the TCLs to OVA indicated that HLA-DR are acting as the dominant restriction elements for these TCLs with minor contribution of HLA-DQ. By use of 187 overlapping synthetic peptides covering the whole sequence of OVA, at least 3 different T cell epitopes were identified.

Topics: Allergens; Amino Acid Sequence; Animals; Cell Line; Chickens; Child, Preschool; Cytokines; Dermatitis, Atopic; Epitope Mapping; Epitopes, T-Lymphocyte; Female; Food Hypersensitivity; HLA Antigens; Humans; Immunoglobulin E; Infant; Lymphocyte Activation; Male; Molecular Sequence Data; Ovalbumin; Peptide Fragments; T-Lymphocytes

Morphologic and immunologic changes in the gut mucosa of food-hypersensitive mice, from a study model generated by feeding ovalbumin (OVA) to female BALB/c mice after intraperitoneal injection of cyclophosphamide (CY), were investigated in an effort to clarify the mechanisms of food-sensitive enteropathy. Villous atrophy, crypt hyperplasia, and increased numbers of intraepithelial lymphocytes (IEL) were confirmed in the antigen-challenged OVA-sensitive mice as seen in food-sensitive enteropathy in humans, whereas no significant morphologic changes were observed in the nontreated control group or groups treated with OVA or CY alone. IEL and lamina propria lymphocytes (LPL) were isolated from the intestinal mucosa before and after the antigen challenge, and surface markers were analyzed by FACScan. After the antigen challenge, the numbers of CD8+ cells increased among the IEL, and the occurrence of both CD4+ and CD8+ cells increased among the LPL. The numbers of Thy-1+ cells and TCR- alpha/beta + cells increased among both the IEL and LPL, and LFA-1 expression was enhanced in both of these lymphocyte populations. The proliferative response of IEL and LPL to OVA increased in a dose-dependent manner after the antigen challenge in the OVA-sensitive mouse model. These results indicate that IEL and LPL, possibly those that have migrated from peripheral blood, are activated by orally administered antigens and cause mucosal damage in the food-sensitive enteropathy.

Topics: Animals; Antigens; Atrophy; Disease Models, Animal; Female; Food Hypersensitivity; Hyperplasia; In Vitro Techniques; Intestinal Diseases; Intestinal Mucosa; Jejunum; Lymphocyte Activation; Mice; Mice, Inbred BALB C; Ovalbumin; T-Lymphocyte Subsets

In the pathogenesis of allergic reactions, T cells and cytokines play a major role. However, characterizations of food allergen-specific T cells are very limited. In this study, we screened the peripheral blood mononuclear cells (PBMC) of 14 patients for reactivity to ovomucoid (Gal d I), the major hen's egg allergen, and ovalbumin (Gal d II). Cell lines and clones specific to ovomucoid were generated from PBMC of four egg-allergic subjects, in order to study antigen domain specificity and cell cytokine production profiles. The results demonstrated, firstly, that egg-allergic patients respond to ovomucoid rather than to ovalbumin, and, secondly, that antigen specificity is predominantly directed toward the second and third domains of ovomucoid. The T-cell cytokine message was characterized by reverse transcriptase polymerase chain reaction (RT-PCR). Cell lines and clones from all four patients consistently expressed interleukin (IL)-5. IL-4, IL-13, and interferon-gamma were found to be expressed only by certain lines or clones. This observation suggests a central pathogenic role for IL-5 in food allergy-related symptoms.

Topics: Adolescent; Adult; Allergens; Cell Division; Cell Line; Child; Child, Preschool; Clone Cells; Cytokines; Eggs; Epitopes; Female; Food Hypersensitivity; Humans; Male; Ovalbumin; Ovomucin; Polymerase Chain Reaction; RNA, Messenger; T-Lymphocytes

Proteins of hen egg whites are common ingredients in food and difficult to eliminate. Allergens of egg white induce allergic symptoms among relatively high numbers of patients suffering from food allergy. B cell epitopes to hen egg white major allergens have been reported. Considering that IgE antibody formation is mostly T cell dependent, the study of T cell epitopes is essential for both T cell dependent and independent IgE response.. Little information on T cell epitopes recognizing food allergens has been reported. T cell responses to hen egg white allergens and two synthetic OA peptides located at amino acid residues No. 105-122 and 323-339 were investigated.. Peripheral blood mononuclear cells from hen egg allergic patients were investigated. Various allergens of hen egg white were used for stimulation. Primary proliferation responses were detected followed by the generation of long-term cultures which were examined for their specificity, phenotype, cytokine profile and IgE production. The allergen specific T cell lines were mapped using a panel of 13 synthetic peptides of ovalbumin.. Human T cells recognizing ovomucoid, lysozyme and ovalbumin epitope 105-122 are reported for the first time. The cell lines were enriched CD4+/CD8+ T cells (CD2+ > 95%). Ovomucoid and ovalbumin induced IgE synthesis by a small fraction of B cells (1%) present in the ovalbumin and ovomucoid specific T cell lines.. Human T cells recognized several egg white allergens and epitopes within the ovalbumin molecule. Specific IgE was produced in cultures stimulated with ovalbumin and ovomucoid. OA peptides 105-122 and 323-339 have no affinity to the specific IgE of the two patients; an observation which could be of particular interest regarding the mechanisms of peptide-based immunotherapy.

Topics: Cell Line; Cells, Cultured; Cytokines; Eggs; Epitopes; Food Hypersensitivity; Humans; Immunoglobulin E; Immunophenotyping; Leukocytes, Mononuclear; Lymphocyte Activation; Male; Muramidase; Ovalbumin; Ovomucin; Peptide Fragments; Radioallergosorbent Test; T-Lymphocytes

Approximately 40 to 80% of egg-allergic children outgrow egg allergy after 2 to 5 years.. To detail the immunologic mechanisms involved in the development of tolerance to egg proteins, the balance between interleukin 4 (IL4) and interferon-gamma (IFN-gamma) synthesis in patients with active atopic dermatitis allergic to hen egg and in those outgrowing hen egg allergy was evaluated.. A marked increase in IL4 and a decrease in IFN-gamma synthesis by peripheral blood lymphocytes following ovalbumin (OVA) specific in vitro stimulation was observed in active atopic dermatitis. In contrast, OVA-induced IL4 synthesis in patients in remission was comparable to that in normal individuals. An intriguing finding was higher production of IFN-gamma by lymphocytes from ovalbumin-insensitive patients in remission as compared to normal individuals following antigen stimulation, although cell proliferation in OVA-stimulated lymphocytes was reduced in patients during remission.. OVA antigen may be capable of inducing a population of Th1-type cells to produce cytokines such as IFN-gamma, resulting in suppression of Th2-type responses, i.e. IL4 secretion. We speculate that the changes in the balance of relevant antigen-induced cytokine synthesis seen in such patients may be causally associated with the improvement in their clinical status.

Topics: Antigens; Child, Preschool; Cytokines; Dermatitis, Atopic; Down-Regulation; Egg Proteins; Eggs; Female; Food Hypersensitivity; Humans; Immune Tolerance; Infant; Interferon-gamma; Interleukin-4; Lymphocytes; Male; Ovalbumin; Up-Regulation

Eicosanoids are involved in the mediation of inflammatory and allergic processes in the gut. In order to evaluate a potential beneficial effect of the diet, the effect of mediators of inflammation and of a sensitization against egg albumin on anion secretion across the colon was tested using rats fed on a diet containing 15% fish oil as compared to 15% olive oil as donor animals. Feeding on a fish oil diet significantly reduced the response to bradykinin or phospholipase C, known agonist of prostaglandin-induced secretion, by about 50%. The increase in short-circuit current (Isc) induced by the phospholipase A2 stimulator, melittin, or by distension of the gut wall were only insignificantly inhibited by 15-30%. Administration of egg albumin to the mucosas from animals sensitized against egg albumin induced an indomethacin- and tetrodotoxin-sensitive increase in Isc. This response was, however, only insignificantly (30%) reduced by the fish oil diet. In conclusion, the effect of fish oil diet depends on the stimulus used for activation of prostaglandin release. This suggested that different pools of arachidonic acid are differentially affected by the diet or that certain stimuli for phospholipases are strong enough to overcome the effect of a reduced substrate availability. Consequently, a diet rich in polyunsaturated n-3 fatty acids may only play an adjuvant role for the therapy of inflammatory or allergic intestinal diseases.

Topics: Acid-Base Equilibrium; Animals; Eicosanoids; Female; Fish Oils; Food Hypersensitivity; Inflammation Mediators; Intestinal Secretions; Ovalbumin; Prostaglandins; Rats; Rats, Inbred Strains

We studied the binding activities of IgE, IgG and IgA antibodies in patients with allergy to hen's egg white against two different ovalbumin (OVA) preparations, which were physically or chemically denatured OVA and enzyme-digested OVA fragments. The binding activities of IgE antibodies to these OVA preparations with those of IgG or IgA antibodies were compared. It was found that the binding activities of IgE antibodies to denatured OVA by treatment with dithiothreitol, urea or hydrochloric acid were similar to those of IgG or IgA antibodies. In contrast, the binding activities of IgE antibodies to heat-denatured OVA or by treatment with sodium hydroxide at pH 11.0 were different from those of IgG or IgA antibodies to these denatured OVA. Furthermore, we found that the binding activities of anti-OVA antibodies in sera from patients with allergy to hen's egg white against fragmented OVA were different between IgE antibodies and IgG or IgA antibodies. Thus, it can be concluded that IgE antibodies to OVA in sera from patients with allergy to egg white differ from IgG or IgA antibodies in respect to binding activities against different preparations of denatured or fragmented OVA, probably due to differences in fine specificities of these antibodies against OVA.

Topics: Allergens; Antibodies; Antibodies, Anti-Idiotypic; Antibody Specificity; Binding Sites, Antibody; Child; Child, Preschool; Egg White; Electrophoresis, Polyacrylamide Gel; Epitopes; Female; Food Hypersensitivity; Humans; Immunoglobulin A; Immunoglobulin E; Immunoglobulin G; Infant; Male; Ovalbumin; Peptide Fragments; Protein Denaturation; Sodium Dodecyl Sulfate

An ovalbumin (OVA)-specific T cell line (TCL) was established from a patient with hen egg allergy. The TCL was CD4+, expressed alpha beta T cell receptor, and recognized OVA presented by HLA-DR10. Based on the response of the TCL to synthetic OVA peptides, it was found that the TCL recognized OVA 323-339, which is a major T cell epitope presented by murine I-Ad. The TCL secreted high levels of IL-5, but undetectable amounts of IL-2, interferon-gamma, and IL-4 when stimulated with OVA or the OVA 323-339 peptide. Since IL-5 is an important growth and chemotactic factor for eosinophils, it is possible that these OVA 323-339-specific T cells can contribute to human egg allergy. To our knowledge, this is the first demonstration of food allergen-specific TCL establishment and identification of a T cell epitope possibly related to the allergic reaction to food antigens. An analog peptide of the OVA 323-339 which is known to strongly bind to I-Ad partially inhibited the response of the TCL to OVA 323-339 presented by HLA-DR10, raising the possibility of peptide-based immunotherapy of food allergy.

Topics: Amino Acid Sequence; Animals; Cell Line; Cell Line, Transformed; Chickens; Child, Preschool; Cytokines; Egg White; Epitopes; Food Hypersensitivity; HLA-D Antigens; Humans; Lymphocyte Activation; Molecular Sequence Data; Ovalbumin; T-Lymphocytes

In order to clarify the mechanism of food-antigen recognition, the proliferative responses of peripheral blood mononuclear cells (PBMCs) to native, heat-denatured or pepsin-treated ovalbumin (OA) were investigated in 16 hen's egg-sensitive patients with atopic dermatitis (AD). Seven of them had hypersensitivity to boiled hen's egg and others had not. The responses of PBMCs to heat-denatured OA were lower than those to native OA in the patients without hypersensitivity to boiled hen's egg. However, there were no differences of the responses of PBMCs between heat-denatured OA and native OA in the patients with hypersensitivity to boiled hen's egg. Moreover, the reduction of the responses of PBMCs to pepsin-treated OA was recognized in six out of seven patients. The primary structure of OA did not change by heating or pepsin treatment according to sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). These results suggested that the secondary structure of OA changed in connection with the reduction of the responses of PBMCs to denatured OA. In addition, we demonstrated the suppressive effect of anti-HLA-DR and anti-HLA-DQ monoclonal antibodies on the proliferative response of PBMCs to OA. The results suggested that the proliferative responses of PBMCs to OA were restricted by HLA-DR or HLA-DQ in hen's egg-sensitive patients with AD.

Topics: Antibodies, Monoclonal; Cell Division; Cells, Cultured; Child; Child, Preschool; Dermatitis, Atopic; DNA; Eggs; Electrophoresis, Polyacrylamide Gel; Female; Food Hypersensitivity; HLA Antigens; Hot Temperature; Humans; Hydrolysis; Immunoglobulin E; Indicators and Reagents; Infant; Male; Monocytes; Ovalbumin; Pepsin A; Protein Denaturation

The study was made of aflatoxin B1 effect on guinea pig alimentary anaphylaxis to chicken ovalbumin (CO) and pasteurized cow milk (PCM), of CO-specific IgG antibody levels, some serum indices, sensitivity of the animals to LD50 histamine. In response to aflatoxin B1 alimentary anaphylaxis both to CO and PCM became more severe, lethality of the anaphylaxis to CO being in logarithmic relation to aflatoxin B1 dose (p < 0.05); specific IgG-antibodies to CO grew in number; serum total protein increased against unchanged levels of albumins; the activity of gamma-glutamyltransferase inhibited; histamine shock gained severity, its lethality being logarithmically related to the aflatoxin B1 dose. The discussion covers mechanisms underlying the animal allergic reactivity responses to aflatoxin B1.

Topics: Aflatoxin B1; Anaphylaxis; Animals; Blood Proteins; Chickens; Dietary Proteins; Drug Interactions; Food Hypersensitivity; gamma-Glutamyltransferase; Guinea Pigs; Immunoglobulin G; Lethal Dose 50; Male; Milk Hypersensitivity; Milk Proteins; Ovalbumin

IgE is considered to be involved in immediate hypersensitive reactions (IHR) following egg ingestion. IgE antibody levels to egg-white (EW) antigens in the IHR-positive group (n = 19, mean age +/- SD = 5.2 +/- 4.5 yr) were higher than those in the IHR-negative group (n = 13, mean of age +/- SD = 3.6 +/- 2.2 yr). However, even in the IHR-negative group, some patients showed high IgE to EW. RAST inhibition tests with heat-treated (100 degrees C, 5, 10, and 30 min) egg-white antigens were performed on 13 serum samples from subjects with IHR and 9 serum samples from subjects without IHR. Heat treatment decreased the IgE-binding activity of egg white and it was speculated that IgE from IHR-negative subjects bound to relatively heat-unstable sites of egg-white antigens. Furthermore, we selected IHR-negative subjects (n = 8, mean of age +/- SD = 3.0 +/- 1.7 yr) with higher IgE antibody levels than the lowest limit of IgE to EW of the IHR-positive group and compared IgE to ovomucoid (OM), ovalbumin (OA), conalbumin (CA), and lysozyme (Ly) between these IHR-negative and positive groups. IgE-binding activities to egg-white components, including OA, CA, and Ly but not OM, were significantly decreased with heat treatment. The IHR-negative group showed significantly lower IgE to OM (untreated, 5, 10, 30 min treatment) and 5 min treated OA alone than the IHR-positive group, while no difference was found in IgE to other components between the two groups.(ABSTRACT TRUNCATED AT 250 WORDS)

Topics: Adolescent; Antigen-Antibody Reactions; Antigens; Child; Child, Preschool; Conalbumin; Egg White; Female; Food Hypersensitivity; Humans; Hypersensitivity, Immediate; Immunoglobulin E; Infant; Male; Muramidase; Ovalbumin; Radioallergosorbent Test

In order to clarify the mechanisms of food-sensitive enteropathy, a food hypersensitive model was generated by feeding ovalbumin to female BALB/c mice after intraperitoneal injection of cyclophosphamide and morphological and immunological changes in the gut mucosa were investigated. Villus atrophy, crypt hyperplasia and increased numbers of intra-epithelial lymphocytes (IEL) were confirmed in this model, as seen in food-sensitive enteropathy in humans. Subpopulations of IEL and lamina propria lymphocytes were enumerated by immunohistochemical observation. CD8-positive cells were increased both in epithelium and lamina propria, whereas CD4-positive cells were decreased in lamina propria. We document here that orally administered food antigen actually induces food-sensitive enteropathy and mucosal damage is generated by lymphocytes that infiltrate the intestinal mucosa. We also investigated the effect of feeding an omega-3 fatty acid-enriched diet in this model and found that it was efficient in attenuating mucosal damage.

Topics: Animals; Fatty Acids, Omega-3; Female; Food Hypersensitivity; Intestinal Diseases; Intestinal Mucosa; Lymphocyte Subsets; Lymphocytes; Mice; Mice, Inbred BALB C; Ovalbumin

Effects of cholera toxin B subunits supplemented with 0.1% cholera toxin (CTB*) on systemic IgE antibody responses to ovalbumin (OVA) were examined in BDFI (H-2b/d), Balb/c (H-2d) and C3H (H-2k) mice given OVA intragastrically or intranasally. Two successive doses of OVA intragastrically administered to Balb/c and C3H mice induced little IgE response and resulted in almost complete unresponsiveness to subsequent intraperitoneal challenge with OVA in Al(OH)3, while the intragastric administration to BDF1 mice induced high IgE response and resulted in abrogation of the unresponsiveness to the subsequent challenge. The intranasal administration of OVA induced little IgE response and suppressed response to the subsequent challenge in any strain of mice. On the other hand, two successive doses of intragastric or intranasal OVA together with CTB* enhanced IgE response in all three strains and the subsequent challenge with OVA in Al(OH)3 induced higher IgE responses. These results suggest that CTB* augments IgE response to OVA and abrogates the unresponsiveness when administered orally or intranasally into mice together with OVA, regardless of the H-2 haplotype of the mice.

Topics: Adjuvants, Immunologic; Administration, Intranasal; Administration, Oral; Animals; Cholera Toxin; Dietary Proteins; Female; Food Hypersensitivity; Immune Tolerance; Immunization; Immunoglobulin E; Mice; Mice, Inbred BALB C; Mice, Inbred C3H; Ovalbumin

When attempting to generate mouse monoclonal antibodies to hen's egg ovalbumin, injection of commercially purified ovalbumin resulted in monoclonal antibodies, which when assayed against commercially purified ovalbumin (Gal d I) or ovomucoid (Gal d III), appeared to be specific to both. With the use of high-performance liquid chromatography (HPLC)-repurified ovalbumin and ovomucoid in assay procedures, monoclonal antibodies generated by commercially purified ovalbumin were found to be specific for ovomucoid only. To clarify this phenomenon, mice were serially injected with commercially purified ovalbumin or HPLC-repurified ovalbumin. It was found that most of the antibody response to commercially purified ovalbumin was directed against the minor (< 1%) ovomucoid contaminant and that HPLC-repurified ovalbumin failed to produce antibodies to ovomucoid. Commercially purified ovomucoid resulted in only minimal amounts of antibodies to ovalbumin. Thus when commercially purified ovalbumin is used both for immunization and immunoassay, most of the antibodies produced are actually against the small amount of ovomucoid contaminant, and not ovalbumin. To determine whether ovomucoid is the major antigenic and allergenic egg white protein in human beings, one group of 18 children with egg allergy were skin prick tested with half-log dilutions of egg white extract and diethylaminoethyl cellulose (DEAE)-repurified ovomucoid, ovalbumin, and lysozyme. Ovomucoid mean wheal diameters were significantly greater than wheal diameters in response to ovalbumin, lysozyme, and egg white extract at the three most concentrated of five dilutions tested: 0.01, 0.03, and 0.1 mg/ml (p < 0.01). Serum ovomucoid-specific IgE and IgG antibody concentrations to DEAE-repurified ovomucoid were significantly greater than that to DEAE-repurified ovalbumin (p < 0.05). In a second study, 10 patients with egg allergy and persistent egg hypersensitivity were compared with 11 patients with egg allergy in whom clinical tolerance to egg developed. IgE antibodies to repurified ovomucoid were significantly greater in patients with persistent egg hypersensitivity compared with patients in whom clinical tolerance developed at the time of both initial and follow-up food challenges. In contrast, there were no significant differences in IgE antibody concentrations to repurified ovalbumin in either group at any time. These results suggest that ovomucoid is the immunodominant protein fraction in egg white and

Topics: Adolescent; Allergens; Animals; Antibody Specificity; Antigens; Child; Child, Preschool; Chromatography, High Pressure Liquid; Eggs; Enzyme-Linked Immunosorbent Assay; Food Hypersensitivity; Humans; Immunoglobulin E; Immunoglobulin G; Infant; Mice; Ovalbumin; Ovomucin; Skin Tests

Specific induction of IL2-responsiveness by ovalbumin-stimulated lymphocytes was studied in patients with hen egg allergy. Fluorescence-activated cell sorter analysis of the cells showed that the IL2-absorbing and IL2-responding cells mainly consisted of CD3+2+4+8-45RA+ cells that may act as helper cells for IgE production and/or as effector cells for delayed type hypersensitivity. beta-Chains (P75) of IL2 receptors were involved in ovalbumin-induced IL2 responsiveness of the patients' lymphocytes, whereas the alpha-chains (p55) were expressed on normal lymphocytes stimulated with ovalbumin as well. Adhering mononuclear cells from patients allergic to ovalbumin but not to Dermatophagoides farinae (Df) were pulsed with ovalbumin antigen then added to a T cell-rich population. After five days of culture, we evaluated cell growth for IL-2 sensitivity during an additional 3-day culture in the presence of IL-2. Responder cells from the patients, which were cocultured with ovalbumin-pulsed autologous adhering cells, acquired IL2 responsiveness; whereas, those cultured with Df-pulsed adhering cells did not. This reaction was specific for antigen. The monoclonal antibody to HLA-DQ (Leu 10) and HLA-DP (HLA-DP) frameworks, but not the one to the HLA-DR framework (OKIa1), blocked the antigen presenting cells ability to induce responses. T Cell-rich responder cells depleted of CD4+ cells did not acquire IL2-responsiveness, whereas the depletion of CD8+ cells had no effect. As a whole, the results indicate that DQ-bearing and/or DP-bearing adhering cells have a key function in presenting ovalbumin-antigen to allergen-specific responder T cells that very likely belong to CD4+ subsets.

Topics: Antigen-Presenting Cells; Cell Adhesion; Eggs; Food Hypersensitivity; HLA-DP Antigens; HLA-DQ Antigens; Humans; Immunity, Cellular; Interleukin-2; Lymphocyte Activation; Lymphokines; Ovalbumin; Receptors, Interleukin-2; T-Lymphocyte Subsets; T-Lymphocytes

Monocyte chemotactic activities in supernatants of ovalbumin (OA)-stimulated peripheral blood mononuclear cell (PBMC) cultures were studied in patients with atopic dermatitis (AD) who were sensitive to hen's egg. The monocyte chemotactic activities in hen's egg-sensitive AD patients were significantly higher than those of non-atopic healthy controls and patients with immediate allergic symptoms. However, the monocyte chemotactic activities were not detected in bovine serum albumin-stimulated PBMC culture supernatants in patients with AD who were sensitive to hen's egg, but not to cow's milk. Furthermore, there was significant correlation between the monocyte chemotactic activities and proliferative responses of PBMCs to OA in hen's egg-sensitive AD patients, whereas there was no significant correlation between the monocyte chemotactic activities and radioallergosorbent test values. These results suggest that PBMCs stimulated with food antigens produce monocyte chemotactic factors which relate to the pathogenesis of AD in food-sensitive AD patients and that the pathogenesis of AD may be related to cell-mediated immune responses.

Topics: Adolescent; Adult; Animals; Chemotactic Factors; Chemotaxis; Chickens; Child; Child, Preschool; Dermatitis, Atopic; Female; Food Hypersensitivity; Humans; In Vitro Techniques; Infant; Lymphocyte Activation; Lymphocytes; Male; Monocytes; Ovalbumin; Ovum

Ten ovalbumin (OVA)-specific T cell lines (TCLs) were established from peripheral blood mononuclear cells of 6 patients with hen egg allergy, and the antigen recognition of these TCLs was characterized. Two OVA epitopes were determined by use of 3 synthetic OVA peptides which have been known as murine T cell epitopes. Blocking of antigen-specific T cell proliferation by anti-HLA class II monoclonal antibodies suggest that all 3 HLA class II molecules could act as restriction elements for T cell recognition of OVA. This is the first demonstration of OVA epitopes recognized by T cells in patients with hen egg allergy, as far as we know.

Topics: Cell Line; Child; Child, Preschool; Eggs; Epitopes; Female; Food Hypersensitivity; Humans; Infant; Male; Ovalbumin; T-Lymphocytes

We tested the effect of Tranilast [N-(3',4'-dimethoxycinnamoyl anthranilic acid)], one of the anti-allergic agents, on the induction of interleukin 2 (IL2) responsiveness of lymphocytes from patients with bronchial asthma or hen-egg allergy following stimulation with Dermatophagoides farinae (Df) or ovalbumin (OVA), respectively. Mononuclear cells pretreated with Tranilast for 12 h failed to respond to IL2 following incubation with Df or OVA. Also Tranilast inhibited purified protein derivative (PPD)-induced IL2 responsiveness of normal lymphocytes but not the Con A-induced IL2 responsiveness of normal or allergen-sensitized lymphocytes. These results suggested that Tranilast has some immunosuppressive effect in that it inhibits antigen-induced IL2 responsiveness. Separation of potential target cells of Tranilast disclosed that antigen-presenting adherent cells were more susceptible to Tranilast than IL2-responding T-cell rich populations. Expression of HLA-DR and -DQ antigens but not DP antigens on macrophages, was significantly suppressed by treatment with Tranilast, although Tranilast scarcely decreased HLA class II antigens expression on B-cells. The suppression was overcome by interferon-gamma, which was known as an inducer for class II antigen expression. Taken together, Tranilast may suppress antigen-induced IL2 responsiveness by inhibiting HLA-DR and HLA-DQ antigens on macrophages.

Topics: Adult; Animals; Antigens, Dermatophagoides; Asthma; Child, Preschool; Food Hypersensitivity; Glycoproteins; Histamine H1 Antagonists; HLA-D Antigens; Humans; In Vitro Techniques; Infant; Interleukin-2; Lymphocytes; Macrophages; Mites; ortho-Aminobenzoates; Ovalbumin

We investigated the effect of elimination diets on T cell responses to ovalbumin (OA) in hen's egg-sensitive atopic dermatitis (AD) patients. The proliferative responses of both peripheral blood mononuclear cells (PBMCs) and T cells with monocytes to OA decreased after elimination diets, but those to Candida albicans or phytohemagglutinin (PHA) did not decrease after elimination diets. The proliferative responses of CD4+ T cells with monocytes to OA decreased after elimination diets. In these patients, clinical symptoms of AD improved. These results indicate that T cells, especially CD4+ T cells, respond to food antigens in food-specific lymphocyte responses, and that elimination diets may be able to initiate reduction of the responsiveness of food-sensitive T cells, especially CD4+ T cells. Moreover, the surface marker phenotypes of the T cells responding to OA were analysed. Our results showed that CD4+CD45RA+ T cells tended to increase. The increase in circulating CD4+CD45RA+ T cells might function as systemic suppression against immune responses in the skin.

Topics: Adolescent; Antigens, Surface; CD4-Positive T-Lymphocytes; Cells, Cultured; Child; Child, Preschool; Dermatitis, Atopic; Eggs; Food Hypersensitivity; Humans; Immunophenotyping; Immunosuppression Therapy; Infant; Lymphocyte Activation; Mitogens; Monocytes; Ovalbumin; T-Lymphocytes; T-Lymphocytes, Regulatory

Young piglets weaned onto soya diets frequently develop diarrhoea which may have a dietary and/or immunological component. Piglets abruptly weaned onto soya at 3 weeks of age developed levels of serum IgG anti-soya antibodies almost comparable to those induced by injection with soya protein in adjuvant at 7 weeks. In the piglets primed by feeding, no significant further increase in antibody occurred after subsequent systemic injection. In contrast, secondary responses were observed in age-matched animals, previously primed by injection, and primary responses were obtained in previously naive piglets. The results demonstrate the development of specific unresponsiveness to soya proteins in neonates fed soya, despite the occurrence of an initial vigorous immune response to the fed protein.

Topics: Administration, Oral; Animal Feed; Animals; Antibodies; Antibody Formation; Dietary Proteins; Female; Food Hypersensitivity; Glycine max; Immune Tolerance; Injections, Intraperitoneal; Male; Ovalbumin; Plant Proteins; Plant Proteins, Dietary; Soybean Proteins; Swine; Time Factors; Weaning

The role of T lymphocytes was assessed in patients with food-sensitive atopic dermatitis (AD). T lymphocytes plus monocytes responded well to ovalbumin or bovine serum albumin (BSA) in children with AD who were sensitive to hen's egg or cow's milk compared with healthy children and children with immediate allergic symptoms who are sensitive to hen's egg or cow's milk. The responding cells were shown to be predominantly CD4+ T lymphocytes. Interleukin-2 activity and interferon-gamma concentrations in culture supernatants of ovalbumin-stimulated peripheral blood mononuclear cells (PBMCs) from patients with AD who were sensitive to hen's egg were significantly higher than those of healthy children and patients sensitive to hen's egg with immediate symptoms. Expression of Fc epsilon R II on B lymphocytes in cultures of ovalbumin-stimulated PBMCs from patients with AD was significantly higher than that of healthy children, but it tended to be lower than that of patients with immediate symptoms. These results suggest that, in patients with AD who are food sensitive, CD4+ T lymphocytes stimulated by food antigens secrete lymphokines such as interleukin-2 and interferon-gamma that are secreted from TH1 clones in mice, and express Fc epsilon R II on B lymphocyte that is induced by interleukin-4 secreted from TH2 clones in mice. Taken together, cell-mediated immunity may also occur in addition to IgE-mediated hypersensitivity in patients with food-sensitive AD.

Topics: Antigens; Cell Division; Cells, Cultured; Child; Child, Preschool; Dermatitis, Atopic; Food Hypersensitivity; Humans; Infant; Interferon-gamma; Interleukin-2; Milk Hypersensitivity; Monocytes; Ovalbumin; Radioallergosorbent Test; Receptors, Fc; T-Lymphocytes

Five patients with atopic dermatitis (AD) who were sensitive to hen's egg were observed before and after natural measles virus infection. Within 4 weeks of natural measles virus infection, the eczematous lesions clearly improved in four of the five patients in whom neither offending foods were eliminated, nor anti-allergic drugs, systemic steroids and steroid ointment administered. This was accompanied by reduced proliferative responses of peripheral blood mononuclear cells (PBMCs) to ovalbumin (OA). Another patient showed a transient improvement of AD symptoms, from severe to mild, and thereafter returned to severe accompanied by increased proliferative responses of PBMCs to OA. Radioallergosorbent test (RAST) scores for hen's egg in all five patients did not change in each level in each patient, except the transiently decreased RAST scores for hen's egg in one patient, after the infection. Thus, in patients with AD who are sensitive to food, the improvement of AD symptoms that appeared within 4 weeks of natural measles virus infection was related to reduced proliferative responses of PBMCs to the food antigen following the infection.

Topics: Antigens; Child; Child, Preschool; Dermatitis, Atopic; Double-Blind Method; Eggs; Food Hypersensitivity; Humans; Infant; Leukocytes, Mononuclear; Lymphocyte Activation; Measles; Measles virus; Ovalbumin

Altered intestinal motility and diarrhea are features of food protein-induced intestinal anaphylaxis in the conscious rat. These experiments were performed to determine the mediator(s) responsible for jejunal circular smooth muscle contraction during this response. Hooded-Lister rats were sensitized by intraperitoneal injection of 10-micrograms egg albumin, and controls were sham-sensitized with saline. Fourteen days later the contractility of the circular muscle in jejunal segments (mucosa intact) was examined in standard tissue baths in response to antigen (Ag) or other agents. While control and sensitized tissues contracted in similar fashion in response to stretch, bethanechol, histamine, or 5-hydroxytryptamine (5HT), Ag contracted only the segments of sensitized animals. The contractile response was: (1) specific to the sensitizing Ag, as bovine serum albumin did not induce contraction and (2) could be passively transferred with serum containing specific immunoglobulin E antibody (IgE-Ab). Concanavalin A, which degranulates both mucosal and connective tissue-type mast cells, and compound 48/80, which degranulates only connective tissue-type mast cells produced contractile responses. Ag-induced contraction was significantly inhibited by the mucosal and connective tissue-type mast cell stabilizer doxantrazole, but not the connective tissue mast cell stabilizer disodium cromoglycate. Diphenhydramine and cimetidine together significantly inhibited histamine-induced contraction, but failed to effect the Ag-induced contraction in sensitized tissues. While the contractile response to 5HT was reduced in the presence of methysergide (5HT1-receptor antagonist), cinanserin (5HT2-receptor antagonist), and ICS 205-930 (5HT3-receptor antagonist), only cinanserin significantly inhibited the contractile response to Ag. Indomethacin significantly inhibited Ag-induced contraction. Ag-induced contraction was resistant to atropine and tetrodotoxin.(ABSTRACT TRUNCATED AT 250 WORDS)

Topics: Anaphylaxis; Animals; Cell Degranulation; Food Hypersensitivity; Gastrointestinal Motility; Immunoglobulin E; Jejunum; Mast Cells; Muscle Contraction; Muscle, Smooth; Ovalbumin; Prostaglandins; Rats; Serotonin

An animal study was conducted to elucidate the role of ovalbumin (OA) in the development of eczematous lesions in intrauterine sensitized newborns. Four groups of pregnant guinea pigs were used: group A, immunized by oral administration of 1% OA in drinking water until parturition; group B, immunized by intradermal injection of OA with Freund's complete adjuvant; group C, immunized by both methods; and group D (control), not immunized. The newborn guinea pigs of each group were patch tested with 10% OA in white petrolatum. Positive reactions were seen in the newborns of groups B and C, but not in those in groups A and D. By enzyme-linked immunosorbent assay and passive cutaneous anaphylaxis, a high titre of OA-specific IgG was detected in the group B and C newborns. The number of positive patch test reactions decreased concomitantly with the decline of specific IgG. Histologically, eczematous changes were observed in the positive reaction sites. Many OA antigen-bearing Langerhans cells were found by the immuno-double labelling technique. Immuno-electron microscopic findings revealed the presence of OA antigens as well as IgG molecules on the cytoplasmic membranes of Langerhans cells. Our studies demonstrated that maternal sensitization with OA can induce an eczematous reaction in the newborns to OA patch testing under the presence of high levels of OA-specific IgG in the serum. From these findings it is suggested that IgG plays an essential role in the development of contact hypersensitivity reaction to OA.

Topics: Administration, Oral; Animals; Animals, Newborn; Dermatitis, Contact; Disease Models, Animal; Female; Food Hypersensitivity; Guinea Pigs; Immunity, Maternally-Acquired; Immunoglobulin G; Injections, Intradermal; Male; Ovalbumin; Passive Cutaneous Anaphylaxis; Patch Tests; Pregnancy

Proliferative responses of peripheral blood mononuclear cells and T cells with monocytes to ovalbumin were significantly higher than those of B cells with monocytes to ovalbumin in patients with atopic dermatitis (AD) who were sensitive to hen eggs. The CD4+ T-cell/CD8+ T-cell ratios (the values obtained by dividing the maximum stimulation index of CD4+ T cells with monocytes to ovalbumin by the maximum stimulation index of CD8+ T cells with monocytes to ovalbumin) were significantly higher in AD patients sensitive to hen eggs than in non-atopic healthy controls. The proliferative responses of CD4+ T cells with monocytes to ovalbumin were more intensive than those of CD8+ T cells with monocytes in patients with AD sensitive to hen eggs compared with non-atopic healthy controls. These results suggest that the cells responding to ovalbumin are predominantly CD4+ T cells. However, there was no relationship between the stimulation index of proliferative responses of peripheral blood mononuclear cells to ovalbumin and RAST scores for hen eggs. Thus it is possible that the majority of the CD4+ T cells which respond to ovalbumin are not CD4+ helper T cells for IgE production.

Topics: Adolescent; B-Lymphocytes; CD4-CD8 Ratio; CD4-Positive T-Lymphocytes; Child; Child, Preschool; Dermatitis, Atopic; Eggs; Evaluation Studies as Topic; Female; Food Hypersensitivity; Humans; Immunoglobulin E; Infant; Leukocytes, Mononuclear; Lymphocyte Activation; Male; Monocytes; Ovalbumin; Radioallergosorbent Test; Severity of Illness Index

Experimental studies have shown that N(3',4'-dimethoxycinnamoyl) anthranilic acid (Tranilast) inhibits reaginic antibody-mediated hypersensitivity reactions, and it has been demonstrated to be an effective drug for patients with bronchial asthma. On the other hand, from the nature of the cellular infiltrate seen in eczematous lesions, it appears that some form of cell-mediated immunity may be involved in addition to IgE-mediated immunity in the pathogenesis of atopic dermatitis (AD). Moreover, we have previously reported that the proliferative responses of peripheral blood mononuclear cells (PBMCs) to ovalbumin (OA) or bovine serum albumin (BSA) in children with AD who are sensitive to hen's egg or cow's milk were significantly higher than those of healthy children and hen's egg or cow's milk sensitive children with immediate symptoms. In this study, we have showed that the proliferative responses of PBMCs to OA were dose-dependently inhibited by Tranilast on patients with AD. The responding cells to OA were shown, through separation experiments, to be T cells, and the proliferative responses of T cells to OA were also dose-dependently inhibited by Tranilast. Moreover, the inhibition was thought to occur at the initial stage of the proliferative reactions. These results suggest that Tranilast can be clinically applied to patients with AD.

Topics: Antigens; Child; Child, Preschool; Dermatitis, Atopic; Food; Food Hypersensitivity; Histamine H1 Antagonists; Humans; In Vitro Techniques; Infant; Lymphocyte Activation; ortho-Aminobenzoates; Ovalbumin; Serum Albumin, Bovine

Proliferative responses of cord blood lymphocytes (CBLs) to food antigens and cord blood IgE concentrations were measured in 37 full term newborn infants for the prediction of allergic disorders. In these 37 infants who were followed up for two years, allergic history of the family was found in four (sensitivity 57.1%) and cord blood IgE concentrations were greater than 0.5 IU/ml in three (sensitivity 42.9%) of seven infants who developed allergic disorders. When CBLs were stimulated twice by ovalbumin or bovine serum albumin, the value of the stimulation index in proliferative responses of CBLs to ovalbumin or bovine serum albumin was greater than 1.5 in six (sensitivity 85.7%) of seven infants who developed allergic disorders. The specificity of the responses of CBLs in the prediction of the development of allergic disorders was 93.3%. The proliferative responses of CBLs to food antigens were useful in the prediction of not only development of allergic disorders but also offending allergens. These observations provide further evidence that sensitisation is occurring in utero. This would appear to be increasingly important in the genesis of early atopic problems. As our follow up is only two years, in utero sensitisation is a prediction for the early development of atopic disease but only longer follow up will show whether this holds good for allergic disorders at any age.

Topics: Allergens; Animals; Cattle; Cell Division; Cells, Cultured; Disease Susceptibility; Fetal Blood; Food Hypersensitivity; Humans; Immunoglobulin E; Infant, Newborn; Lymphocytes; Ovalbumin; Prognosis; Serum Albumin

To assess whether dietary antigens play a role in inflammatory bowel disease, 26 monozygotic twin pairs with inflammatory bowel disease and 52 healthy controls were investigated for serum antibodies (IgA, IgG, IgM) against ovalbumin, betalactoglobulin, gliadin, whole yeast (Saccharomyces cerevisiae) and yeast cell wall mannan. The twins were made up of five pairs concordant and nine pairs discordant for Crohn's disease, and two pairs concordant and 10 pairs discordant for ulcerative colitis. Two patients with Crohn's disease had a slight increase in disease activity, the others were in clinical remission. Two striking observations were made: first, individuals with ulcerative colitis were indistinguishable from healthy twins, and controls except for the response to gliadin. Both healthy and diseased twins had higher IgA levels to gliadin than controls. Second, twins who had developed Crohn's disease displayed higher antibody titres towards yeast cell wall mannan in particular, but also to whole yeast (Saccharomyces cerevisiae) of all antibody types (IgA, IgG, and IgM). In contrast, the response to gliadin, ovalbumin, and betalactoglobulin did not differ from healthy twins and was even lower than in the controls. The results argue against an increased systemic antigen presentation caused by an impaired mucosal barrier in the inflammatory bowel disease. Rather, they suggest that yeast cell wall material--that is, mannan, or some antigen rich in mannose and cross reacting with mannan, may play an aetiological role in Crohn's disease, but not in ulcerative colitis. The increases in IgA and IgM, as well as IgG suggest that local and systemic immune systems are selectively activated by antigen(s) present in the cell wall of baker's yeast.

Topics: Adult; Aged; Cell Wall; Food Hypersensitivity; Gliadin; Humans; Immunoglobulin A; Immunoglobulin G; Immunoglobulin M; Inflammatory Bowel Diseases; Lactoglobulins; Mannans; Middle Aged; Ovalbumin; Saccharomyces cerevisiae; Twins, Monozygotic

To delineate the antigenic determinants on ovalbumin (OA) and ovomucoid (OVM) recognized by specific IgG and IgE antibodies in sera from patients with allergy to hen's egg, we studied the binding activities of IgG and IgE antibodies to native OA or OVM and seven different OA or OVM preparations, i.e. heated OA or OVM (100 degrees C for 3 min and 80 degrees C for 30 min) or 0.3 M NaOH, dithiothreitol (DTT), sodium dodecyl sulfate (SDS), 6 M urea or HCl treated OA or OVM. Eight patients with IgE anti-OA antibodies and 12 patients with IgE anti-OVM antibodies were used in these studies. The binding activities of IgG and IgE antibodies to physically or chemically denatured OA were partially decreased compared with those to native OA, whereas IgG and IgE antibodies in sera from patients bound well to denatured OVM with similar binding activities to native OVM. These results strongly suggest that antibodies to OA recognize partially conformational antigenic determinants on OA, whereas antibodies to OVM mainly recognize sequential antigenic determinants on OVM, and that antigenic determinants of OVM recognized by antibodies in sera from patients are more stable than those of OA under these denaturation conditions. In addition, the binding activities of IgE antibodies to denatured OA or OVM were significantly different from those of IgG antibodies to these OA or OVM, suggesting that the specificities of IgE antibodies to OA or OVM may be different from those of IgG antibodies to OA or OVM.

Topics: Adolescent; Animals; Antibody Specificity; Child; Child, Preschool; Eggs; Epitopes; Female; Food Hypersensitivity; Humans; Immunoglobulin E; Immunoglobulin G; Infant; Male; Mice; Ovalbumin; Ovomucin; Protein Denaturation; Rabbits

Topics: Anaphylaxis; Animals; Chickens; Cytochrome P-450 Enzyme System; Food Hypersensitivity; Guinea Pigs; Male; Methylcholanthrene; Microsomes, Liver; Ovalbumin

Topics: Adolescent; Animals; Chickens; Child; Child, Preschool; Dermatitis, Atopic; Diarrhea; Edible Grain; Egg White; Female; Fluoroimmunoassay; Food Hypersensitivity; Humans; Immunoglobulin E; Immunoglobulin Fragments; Immunoglobulin G; Infant; Milk Proteins; Ovalbumin; Radioallergosorbent Test

Allergen-specific B cells were detected by antigen-coated magnetic beads. The frequency of ovalbumin (OVA)-specific B cells was significantly higher in patients with egg white-allergy than in age-matched nonallergic individuals. These B cells, isolated by the immunomagnetic-beads method, produced anti-OVA antibodies of mostly IgM class when they were transformed by Epstein-Barr virus and cultured for about 2 weeks. Based on a chronologic analysis, the increase of OVA-specific B cells was found to precede the increase of IgG and IgE anti-OVA antibodies in the serum. These observations indicated that OVA-binding B cells in the peripheral blood are already committed to producing IgM antibody and probably are the precursors of antibody-forming cells of the IgG or IgE class.

Topics: Adult; B-Lymphocytes; Cells, Cultured; Child, Preschool; Eggs; Food Hypersensitivity; Humans; Immunoglobulin A; Immunoglobulin E; Immunoglobulin G; Immunoglobulin M; Infant; Lymphocyte Activation; Ovalbumin; Rosette Formation; Serum Albumin

The IgE levels and food-allergen-specific IgE- and IgG-antibodies (Ab) to ovalbumin (OA), ovomucoid (OVO) and beta-lactoglobulin (BLG) were determined up to 18 months of age in 163 infants born to women who were atopic. A high (HIGH group) or a low (REDUCED group) intake of hen's egg and cow's milk by the mother during the third trimester gave no significant differences in the concentrations of IgE or in IgE-Ab (OVO, BLG) and IgG-Ab (OA, OVO, BLG). Similarly, a prolongation of the abstention diet to the early lactation period did not influence the immune response. The IgG-Ab levels to all three food allergens decreased significantly (P less than 0.001) in both study groups between birth and 2 months of age, but then increased significantly (P less than 0.001) between 6 and 18 months of age. The presence in serum of IgE-Ab to OVO (greater than or equal to 0.15 PRU/ml) was associated with significantly higher IgG-Ab levels to OVO at 6 months (P less than 0.001) and at 18 months (P less than 0.05). Infants with positive skin-prick tests (SPT) to OA and OVO showed higher IgG-Ab levels at 6 and 18 months of age than did infants with negative SPT reactions to the two egg allergens. This indicates a relation between the IgE- and IgG-Ab response and it also suggests that some individuals are 'high responders' to both types of immunoglobulin isotypes while others are 'low responders'.

Topics: Allergens; Animals; Diet; Eggs; Female; Food Hypersensitivity; Humans; Immunoglobulin E; Immunoglobulin G; Infant; Lactation; Lactoglobulins; Male; Maternal-Fetal Exchange; Milk; Ovalbumin; Ovomucin; Pregnancy

Topics: Child, Preschool; Eggs; Evaluation Studies as Topic; Food Hypersensitivity; Humans; Immunoglobulin E; Measles; Measles Vaccine; Ovalbumin

The effect of chronic dietary antigen challenge on the intestine was examined in sensitized rats. Three groups of Hooded-Lister rats were studied: animals sensitized to egg albumin; sham-sensitized animals; and unmanipulated controls. In sensitized rats, serum immunoglobulin E titers to egg albumin were greater than or equal to 1:64, whereas control and pair-fed rats showed no response. Sensitized rats received egg albumin 1 mg/ml in drinking water and rat chow ad libitum. Pair-fed animals also received egg albumin but were pair-fed with sensitized animals. Controls received water and rat chow ad libitum. Chronic antigen challenge resulted in reduced food intake and weight gain in sensitized animals. When the rats were killed after 9 days of antigen exposure, proximal intestine from experimental animals showed decreased disaccharidase activity, brush-border microvillus surface, area, and villus height. Crypt depth and enterocyte migration rate were increased. Mucosal mast cell involvement was suggested by mast cell proliferation, evidence of mast cell degranulation, and increased serum rat mast cell protease II levels. At the time of death, only sensitized jejunum demonstrated an increase in short-circuit current in Ussing chambers in response to antigen challenge. The findings indicate that chronic antigen exposure leads to intestinal injury, reduced food intake, and diminished weight gain.

Topics: Anaphylaxis; Animals; Antigens; Body Weight; Female; Food Hypersensitivity; Immunization; Immunoglobulin E; Intestinal Diseases; Intestinal Mucosa; Intestine, Small; Mast Cells; Microscopy, Electron; Ovalbumin; Peptide Hydrolases; Rats; Rats, Inbred Strains

Thirty-five egg-sensitive children who received measles immunization without adverse sequelae are described. Thirty-two of the children had a history of immediate hypersensitivity reactions to egg protein, including 22 who developed a generalized reaction after oral exposure to egg. There were also 3 highly allergic children, with immediate hypersensitivity reactions to other food, who, despite having never been exposed to egg, developed large skin prick test wheals to egg white. Measles vaccine was given to all children without prior vaccine skin testing. There were no adverse reactions. It is suggested that measles vaccine can be given to children with a history of generalized or localized urticaria/angioedema on exposure to egg protein without prior skin testing.

Topics: Child, Preschool; Egg Proteins, Dietary; Egg White; Egg Yolk; Female; Food Hypersensitivity; Humans; Hypersensitivity, Immediate; Infant; Male; Measles Vaccine; Milk Proteins; Ovalbumin; Skin Tests

Alterations in myoelectric and motor activity are important features of food protein-induced intestinal anaphylaxis. To determine the mediator(s) involved, rats were sensitized by injection of egg albumin (10 micrograms ip), and controls were sham sensitized with saline. Fourteen days later the contractility of longitudinally oriented jejunal segments (mucosa intact) was examined in standard tissue baths in response to antigen (Ag) or other agents. Although control and sensitized tissues similarly contracted to stretch, bethanechol, histamine, or 5-hydroxytryptamine (5-HT), Ag contracted only sensitized segments. Contractile response 1) was specific to the sensitizing Ag, as bovine serum albumin did not induce contraction, and 2) could be passively transferred with serum containing specific IgE antibody. Mast cell degranulation after Ag challenge was suggested by a significant loss of granulated mast cells in sensitized compared with control rats challenged with Ag. Concanavalin A, which degranulates mucosal and connective tissue-type mast cells, and compound 48/80, which degranulates only connective tissue-type mast cells, produced a contractile response. Ag-induced contraction was significantly inhibited by the mucosal and connective tissue-type mast cell stabilizer doxantrazole (P less than 0.001) and the connective tissue mast cell stabilizer disodium cromoglycate (P less than 0.05). Diphenhydramine and cimetidine together blocked histamine-induced contraction but failed to affect Ag-induced contraction in sensitized tissues.(ABSTRACT TRUNCATED AT 250 WORDS)

Topics: Animals; Chickens; Dietary Proteins; Food Hypersensitivity; Immunoglobulin E; In Vitro Techniques; Intestinal Mucosa; Jejunum; Mast Cells; Muscle Contraction; Muscle, Smooth; Ovalbumin; Rats; Rats, Inbred Strains; Reference Values

Groups of rats previously sensitized systemically on day 0 with a low dose of ovalbumin (OVA) were gavaged daily with ovalbumin or bovine serum albumin or a mixture of both proteins from day 14 to 18. Blood samples were obtained pre- and post-challenge and serum levels of the rat mast cell protease II (RMCPII) determined by immunoassay. Release of this specific mucosal mast cell mediator was only observed in animals challenged with ovalbumin and the initial challenge released levels of RMCPII 15-fold higher than normal resting levels (P less than 0.001). Subsequent daily challenges evoked the release of significantly lower levels of mediator (P less than 0.001 relative to day 14), but with one exception each test group released significantly more RMCPII than the matched control group on each day (P less than 0.001-P = 0.015). An increased uptake of BSA 'bystander' protein was observed when OVA-sensitized animals were repeatedly gavage-challenged with OVA but there was no correlation with the release of RMCPII mediator. After a 9-day rest period the levels of RMCPII released 6 hr post-challenge on day 26 were still significantly lower (P = 0.004) than the levels of mediator released on day 14. In contrast, animals not previously challenged were still capable of releasing high levels of mediator at the time of first mucosal contact. The levels of RMCPII detected in the serum after enteral protein antigen challenge never exceeded 6000 ng/ml and were lower than those previously observed in parasitized rats following intravenous antigen challenge.

Topics: Anaphylaxis; Animals; Antigens; Chymases; Disease Models, Animal; Food Hypersensitivity; Immunization; Injections, Intraperitoneal; Intestinal Mucosa; Intubation, Gastrointestinal; Mast Cells; Ovalbumin; Rats; Rats, Inbred Strains; Serine Endopeptidases

We studied antigen-specific IgA production from the peripheral lymphocytes of children with hen-egg allergy by using the method of indirect plaque forming cell (PFC) assay. The number of OVA-specific IgA-PFC generated from the patient lymphocytes was low (6 +/- 3/7 X 10(4) non T cells) in comparison with age-matched normal children (112 +/- 18/7 X 10(4) non T cells). The numbers of OVA-specific IgG-PFC and IgM-PFC generated from the patient lymphocytes were not so different from those of the age-matched normal children. This indicates that the activity of OVA-specific IgA-antibody production was reduced in patients with hen-egg allergy.

Topics: Child, Preschool; Eggs; Female; Food Hypersensitivity; Humans; Immunoglobulin A; Immunoglobulin M; Lymphocytes; Male; Ovalbumin

Breast milk samples were collected from 152 women during the first week after delivery. The levels of IgG and IgA antibodies to beta-lactoglobulin, ovalbumin and gliadin were assessed with an enzyme-linked immunosorbent assay (ELISA). The breast milk antibody levels did not differ significantly between mothers on a strictly cow's milk and egg-free diet, and mothers taking these foods. Moreover, the colostral food antibody levels did not differ significantly between atopic and non-atopic mothers. Neither was there any correlation between the colostral antibody levels and the development of atopic disease in the baby. I conclude that maternal antigen avoidance during late pregnancy does not affect the food antibody levels in colostrum. High levels of food antibodies in a colostrum sample seem not to offer protection against food allergy in the child.

Topics: Animals; Colostrum; Diet; Food Hypersensitivity; Gliadin; Humans; Hypersensitivity; Infant; Infant, Newborn; Lactoglobulins; Milk; Milk, Human; Ovalbumin; Prospective Studies

Increased gastrointestinal absorption of intact antigen with systemic immunization has been considered a major etiologic factor in the development of food sensitivity. We attempted to test this hypothesis in infants with suspected food protein-induced entercolitis by measuring serum ovalbumin (OVA) concentrations after ingestion of egg white (prior to the performance of good challenges to establish this diagnosis). We first noted significant underestimation of serum OVA concentrations in the presence of even low serum anti-OVA antibody concentrations (greater than 1:12). Next, using selected noninhibitory sera, we found that all infants studied absorbed some OVA, there was no correlation between serum OVA levels and age (3-11 months), and there was no significant difference between serum OVA concentrations in infants who subsequently had positive oral food challenge responses (120 +/- 67 ng/ml) and a matched group with negative challenges (102 +/- 80). These data do not support the hypothesis that "intestinal closure" (antigen exclusion) occurs in the neonatal period or the role of antigen absorption as the major etiological factor in the development of food sensitivity. Better methods of quantitating macromolecular absorption must be developed before the role of antigen absorption in food sensitivity can be assessed. Of note, urinary excretion of intact OVA also occurred. This varied greatly from one voiding to the next and continuing for at least 13 hr after ingestion.

Topics: Antibodies; Antigens; Dietary Proteins; Enterocolitis; Female; Food Hypersensitivity; Humans; Immunization; Infant; Intestinal Absorption; Male; Ovalbumin

Ovalbumin (OA) and specific antibodies to OA were evaluated by ELISA in 92 colostrum, 366 mature milk and 12 cord blood samples. Anti-OA IgG antibody titers in colostrum were lower than those in cord blood and were not detectable in several samples. Anti-OA IgM antibody titers in colostrum were comparatively high and decreased on the days following postpartum. Anti-OA IgA antibody titers were the highest in the colostrum and decreased on the following days, but remained constant in mature milk. On the other hand, OA was detected in 27.2% of colostrum and 17.2% of mature milk samples, in concentrations from 0.5-59.0 ng/ml. In mature milk, the positive rates of OA were significantly elevated by egg ingestion by the mothers. OA in mature milk from allergic mothers tended to be more readily detectable than that from non-allergic mothers. These results suggest that avoidance of the specific food antigens by mothers is important for therapy and prevention on allergic disease in their breast-fed infants.

Topics: Breast Feeding; Eggs; Enzyme-Linked Immunosorbent Assay; Food Hypersensitivity; Humans; Immunoglobulin A; Immunoglobulin G; Immunoglobulin M; Milk, Human; Ovalbumin

The role of IgG subclasses and IgG subclass antibodies in atopic disease is controversial. Serum IgG and IgG subclass (IgG1-4) antibodies to the two dietary antigens ovalbumin (OA) and beta-lactoglobulin (BLG) were measured with ELISA-methods in 16 patients with mild or moderate atopic dermatitis (AD) and healthy controls. The IgG antibodies were measured in 31 patients with previous AD and controls. The IgG subclass antibodies to OA and BLG showed predominance of IgG4 and IgG1 for both patients and controls. The levels of IgG and IgG subclass antibodies to OA did not differ between the groups, but the levels of IgG and IgG4 anti-BLG antibody were higher in patients with active AD than in controls. The antibody levels did not correlate with severity of disease or with a history of food allergy/intolerance. IgG4 antibodies to dietary antigens may be elevated in AD, but the diagnostic significance of IgG subclass antibody measurement is limited.

Topics: Animals; Antigens; Dermatitis, Atopic; Food Hypersensitivity; Humans; Immunoglobulin E; Immunoglobulin G; Lactoglobulins; Milk; Ovalbumin

Topics: Allergens; Antibody Formation; Antibody Specificity; Child; Child, Preschool; Enzyme-Linked Immunosorbent Assay; Food Hypersensitivity; Humans; Immunoglobulin G; Infant; Lactoglobulins; Ovalbumin; Reference Values

Serum concentrations of IgE, IgG1 and IgG4 antibodies to ovalbumin and basophil sensitivity to ovalbumin were compared with the results of a titrated oral provocation test with ovalbumin in 27 children sensitive to hen's egg white, of whom 17 responded with an immediate hypersensitivity reaction. Neither the serum level of IgE, IgG1 and IgG4 antibodies to ovalbumin nor a positive histamine release test predicted the clinical relevance of ovalbumin sensitivity. The children with a positive challenge test had a significantly higher IgE/IgE4 ratio and tended to be younger and to have higher serum IgE levels and a higher IgG1/IgG4 ratio than those with a negative challenge test. We conclude that an oral provocation test is necessary to confirm the diagnosis of food allergy.

Topics: Child; Child, Preschool; Egg White; Enzyme-Linked Immunosorbent Assay; Female; Food Hypersensitivity; Histamine Release; Humans; Hypersensitivity, Immediate; Immunoglobulin E; Immunoglobulin G; Infant; Male; Ovalbumin

Intestinal motility was examined in an animal model of intestinal anaphylaxis. Hooded-Lister rats were sensitized (S) by intraperitoneal injection of 10 micrograms egg albumin (Ag) and compared with sham-sensitized controls (C). Seven days later three bipolar jejunal electrodes and a jejunostomy tube for motility recording and Ag administration were implanted. On day 14 intestinal myoelectric and motor activity were measured in fasted animals before and after intraluminal administration of either 10 mg egg albumin in 0.5 ml saline, 10 mg bovine serum albumin (BSA) in 0.5 ml saline, or placebo (P) challenge with 0.5 ml saline. Specific immunoglobulin E serum titers were greater than or equal to 1:64 in S animals, whereas C animals showed no response. None of the C animals challenged with P or Ag and none of the S animals challenged with P or BSA defecated after challenge, but all the S animals challenged with Ag developed diarrhea (P less than 0.001). In S animals challenged with Ag, the fasting motility pattern was disrupted, the migrating motor complex was abolished (P = 0.002), and the frequency of aborally propagating clustered contractions was increased (P less than 0.01). In this animal model an immune-mediated reaction to food protein was associated with diarrhea and altered intestinal myoelectric and motor activity.

Topics: Anaphylaxis; Animals; Chickens; Disease Models, Animal; Fasting; Food Hypersensitivity; Gastrointestinal Motility; Jejunum; Muscle, Smooth; Ovalbumin; Rats; Rats, Inbred Strains; Reference Values

Topics: Food Hypersensitivity; Humans; Intestine, Small; Ovalbumin; Permeability

Topics: Antigens; Breast Feeding; Caseins; Dermatitis, Atopic; Egg Proteins; Food Hypersensitivity; Humans; Immunoglobulins; Infant; Infant, Newborn; Lactalbumin; Lactoglobulins; Milk, Human; Ovalbumin

Topics: Animals; Autoantibodies; Child; Child, Preschool; False Negative Reactions; Food Hypersensitivity; Humans; Immunoglobulin E; Infant; Milk; Ovalbumin; Radioallergosorbent Test; Radioimmunosorbent Test

Topics: Animals; Cells, Cultured; Chickens; Child; Eggs; Food Hypersensitivity; Humans; Immunity, Cellular; Lymphocyte Activation; Ovalbumin; T-Lymphocytes

Intestinal anaphylaxis is associated with disturbances in gut function that are antigen-specific and dependent on mast cell degranulation. Using an animal model of intestinal anaphylaxis, we have correlated alterations in water and electrolyte transport, associated with intraluminal challenge, with specific intestinal mucosal mast cell degranulation by following systemic as well as local release of rat mast cell protease II. This protease is specific for intestinal mucosal mast cells and is known to selectively attack type IV collagen, which is found in basement membranes. Intraluminal antigen challenge in sensitized animals dramatically increased serum and intraluminal levels of rat mast cell protease II. Serum levels continued to rise throughout the duration of antigen challenge. Although light microscopy of challenged intestine demonstrated little distortion of mucosal architecture, ultrastructural examination revealed significant disruption to the basement membrane and underlying collagenous matrix of the intestinal mucosa. Our findings indicate that during mucosal immunoglobulin E-mediated reactions, rat mast cell protease II is released and is associated with ultrastructural changes in the intestinal mucosa. The systemic appearance of this specific protease provides a serum marker of intestinal anaphylaxis.

Topics: Anaphylaxis; Animals; Chymases; Female; Food Hypersensitivity; Immunoglobulin E; Intestinal Absorption; Intestinal Mucosa; Mast Cells; Microscopy, Electron; Ovalbumin; Rats; Serine Endopeptidases; Water-Electrolyte Balance

We have confirmed previous observations that intestinal anaphylaxis induced in rats previously sensitized to ovalbumin (OVA) is associated with an increased uptake of an unrelated 'bystander' protein, bovine serum albumin (BSA) fed 1 hr previously. In this study, this enhanced protein uptake was associated with an increased lactulose/rhamnose excretion ratio after administration of these sugars, although there was no correlation between the two measurements. One hour after antigen challenge the serum levels of rat mast-cell protease II (RMCPII), a specific marker for mucosal mast-cell secretion, were significantly higher than both the pre-challenge levels and those of sham-challenged controls (P less than 0.002). There was a significant positive correlation between the serum levels of RMCPII and the lactulose/rhamnose excretion ratios (P less than 0.05), but no such correlation existed between RMCPII and BSA levels in the challenged rats. In other studies the urinary lactulose/rhamnose ratios of rats with cetrimide-induced gut damage were found to be significantly increased, although BSA uptake into the serum remained unaltered. We conclude that there is no simple correlation between gut permeation of low-molecular weight sugars and and the uptake of macromolecular proteins.

Topics: Animals; Chymases; Disaccharides; Female; Food Hypersensitivity; Intestinal Absorption; Intestinal Mucosa; Lactulose; Male; Mast Cells; Ovalbumin; Permeability; Rats; Rats, Inbred Strains; Rhamnose; Serine Endopeptidases; Serum Albumin, Bovine

Topics: Food Hypersensitivity; Humans; Lymphocyte Activation; ortho-Aminobenzoates; Ovalbumin; Serum Albumin, Bovine

Experiments in order to induce food allergy were carried out in guinea pigs. The sensitization with egg albumin, pasteurized cow milk and bovine serum albumin provoked anaphylactic shock. The passive cutaneous anaphylaxis, serum antibodies, liver cytochrome P-450 concentration and the anaphylactic shock were determined. Some correlation between the mortality, anaphylactic antibodies and cytochrome P-450 monooxygenase system was established. The morphology of the jejunal mucosa, the activities of the 5 disaccharidases, the number of immunoglobulin secreting cells (Ig SC) and the mastocytes were investigated in 35 patients with food allergy. Normal mucosa was found in 28 cases as well as a significant decrease of the lactase, sucrase and trehalase activities. An increase of IgM and IgG secreting cells and of mastocytes, different electron microscopic changes in the enterocytes (an increased number of lysosomes, appearance of vesicles in cytoplasma, shortening, enlargement and uneven distribution of microvilli) as well as symptoms of functional activity in the plasmocytes and some others were also revealed. The experimental model obtained is similar to that one in humans according to the enteral way of sensitization the high selectivity of the allergic reaction which is of reagin type as the immunoglobulin changes are involved.

Topics: Animals; Antibody Specificity; Antigen-Antibody Complex; Antigens; Disaccharidases; Food Hypersensitivity; Guinea Pigs; Humans; Immunoglobulin E; Immunoglobulins; Intestinal Mucosa; Microscopy, Electron; Ovalbumin; Radioimmunoassay; Rats; Rats, Inbred Strains; Serum Albumin, Bovine

We have examined the role of Ts cels and APC in regulating the tolerance of systemic DTH in mice fed OVA. Oral tolerance to OVA was prevented by eliminating Ts with dGuo and by treating mice with anti-I-J antiserum. In addition, activating the reticuloendothelial system (RES) with oestradiol, muramyl dipeptide (MDP) or GvHR prevented the induction of tolerance. Further studies showed that prevention of oral tolerance correlated with the ability to enhance APC activity and that oestradiol also abrogated the induction of Ts after feeding OVA. Our results show that the tolerance of systemic DTH in mice fed OVA reflects complex interactions between APC and Ts and suggest that defects in these regulatory events may be responsible for clinical food sensitive enteropathy.

Topics: Administration, Oral; Animals; Antigen-Presenting Cells; Deoxyguanosine; Dietary Proteins; Food Hypersensitivity; Graft vs Host Reaction; Hypersensitivity, Delayed; Immune Tolerance; Mice; Mononuclear Phagocyte System; Ovalbumin; T-Lymphocytes, Regulatory

Topics: Animals; Child; Child, Preschool; Eggs; Food Hypersensitivity; Humans; Infant; Lymphocytes; Milk; Ovalbumin; Serum Albumin, Bovine

Serum antibodies of the IgG type from rabbits immunized with food antigens (beta-lactoglobulin, ovalbumin and gliadin) have been quantified using a fluorescence-linked immunosorbent assay (FLISA), with the antigens adsorbed as round spots (about 8 mm in diameter) on glass or plastic microscope slides. The indirect immunofluorescence intensities were determined using a microscope fluorometer, and compared to enzyme-linked immunosorbent assay (ELISA) in microtiter plates and diffusion-in-gel-ELISA (DIG-ELISA) in plastic petri dishes. It was found that FLISA in general became more sensitive when the antigens had been adsorbed onto a plastic (Nunclon) than onto a glass surface. When the antigens were adsorbed to the plastic slides, the relative sensitivity order (maximum serum dilution) of the assays was in general the following, ELISA greater than FLISA greater than DIG-ELISA. The fluorescence-linked method appeared to require equal or less antigen and conjugated antiserum per sample. Due to the visual inspection of the surface, inhomogeneities of the antigen-coating could be readily discovered and evaluated by several measurements within the field of antigen-antibody reaction. It is proposed that spot FLISA may be an alternative to ELISA especially when the amount of antigen or antiserum is limited.

Topics: Allergens; Animals; Antibodies; Fluorescent Antibody Technique; Food Hypersensitivity; Gliadin; Immunization; Immunosorbent Techniques; Lactoglobulins; Ovalbumin; Rabbits

In IgA glomerulonephritis (GN), the pathogenic role of IgA is well documented, but the specificity of these IgA is unknown. Cases of celiac disease associated with IgA GN have been reported and led us to investigate the role of gliadin sensitivity. We measured IgA, IgG and IgM antibodies to gliadin, beta-lactoglobulin and ovalbumin by ELISA (results expressed as optical density; OD) in 27 patients with primary IgA GN, 14 with membranous GN (MGN), 21 with idiopathic nephrotic syndrome (INS) and 21 healthy controls. The normal value for antigliadin IgA was less than 0.650 OD. 19/27 patients with IgA GN had a raised level versus 2/14 in MGN and 2/21 INS (p less than 0.001: IgA GN vs. MGN, INS and controls). Antibodies to beta-lactoglobulin were rarely found and were not more frequent in IgA GN. Cross-reactivity with reticulin was investigated in 16 patients who were serum-positive for IgA antigliadin: no reticulin antibodies were detected by immunofluorescence. Antigliadin IgA are of diagnostic value for distinguishing IgA GN from other GN, with a sensitivity of 70%, a specificity of 89%, a positive predictive value of 83% and a negative one of 79%.

Topics: Adult; Antibody Specificity; Celiac Disease; Dietary Proteins; Enzyme-Linked Immunosorbent Assay; Female; Food Hypersensitivity; Gliadin; Glomerulonephritis, IGA; Humans; Immunoglobulin A; Lactoglobulins; Male; Ovalbumin; Plant Proteins

The radioallergosorbent test (RAST) and RAST inhibition test were used to examine cross-allergenicity amongst the major hen's egg-white and egg-yolk proteins. Using ovalbumin as a reference allergen to compare cross-reactivity, it was apparent that the proteins conalbumin, ovomucoid and lysozyme substantially inhibited binding to ovalbumin discs of IgE in the sera of patients clinically hypersensitive to egg. The converse situation with conalbumin, ovomucoid and lysozyme on the discs and ovalbumin as the inhibitor also resulted in significantly decreased levels of IgE binding to the proteins on the discs. It was also demonstrated that cross-reactions occurred between ovalbumin and the yolk protein, apovitellenin I. Cross-reaction was also observed surprisingly when egg lysozyme was on the disc and the milk protein allergen alpha-lactalbumin was used as the inhibitor. The demonstration of cross-reaction between all of these proteins may signify that there are a number of common allergenic determinants on these egg proteins, thus providing a molecular basis for the phenomenon of cross-reactivity.

Topics: Adolescent; Adult; Apoproteins; Child; Child, Preschool; Cross Reactions; Egg Proteins; Egg Proteins, Dietary; Food Hypersensitivity; Humans; Lactalbumin; Muramidase; Ovalbumin

Topics: Adult; Antigen-Antibody Complex; Asthma; Eggs; Female; Food Hypersensitivity; Humans; Immunoglobulin G; Male; Middle Aged; Ovalbumin; Radioallergosorbent Test

In this study we have examined whether differences between mouse strains in the induction of tolerance after feeding ovalbumin (OVA) are due to differences in intestinal processing of OVA or are determined by the systemic immune system. Compared with major histocompatibility complex (MHC)-congenic BALB/c mice, BALB/B mice develop much less tolerance of systemic delayed-type hypersensitivity (DTH) and humoral immunity after feeding OVA and this defect is also expressed partially in (BALB/B x BALB/c)F1 animals. Serum taken from either BALB/c or BALB/B mice fed OVA 1 h before produced significant suppression of systemic DTH responses in BALB/c, but not in BALB/B mice. Although OVA-fed BALB/B serum was slightly less tolerogenic than BALB/c serum, we conclude that the defective induction of oral tolerance in BALB/B mice is due primarily to a MHC-influenced defect with the immune system. These findings support the idea that clinical food-sensitive enteropathy reflects an immune response gene-controlled defect in tolerance to dietary proteins.

Topics: Administration, Oral; Animals; Dietary Proteins; Food Hypersensitivity; Hypersensitivity, Delayed; Immune Tolerance; Immunologic Deficiency Syndromes; Intestines; Major Histocompatibility Complex; Mice; Mice, Inbred BALB C; Mice, Inbred Strains; Ovalbumin

Topics: Animals; Diphenhydramine; DNA; Epithelial Cells; Epithelium; Food Hypersensitivity; Histamine; In Vitro Techniques; Intestinal Absorption; Intestinal Mucosa; Mast Cells; Ovalbumin; Proteins; Rats; Sucrase; Tetrodotoxin; Thioxanthenes; Water-Electrolyte Balance; Xanthones

We investigated the transfer of bovine serum 125I-albumin (125I-BSA), bovine 125I-gamma-globulin (125I-BGG), 125I-ovalbumin (125I-OVA), and 125I-beta-lactoglobulin (125I-BLG) from the blood into the milk of lactating mice. Equal amounts (by weight) of the radiolabeled proteins were injected intravenously into mice 1 wk postpartum. Total radioactivity, trichloroacetic acid-precipitable radioactivity, and specifically immunoprecipitable radioactivity were measured in serum, mammary gland homogenate, and milk. Clearance of immunoreactive OVA (iOVA) and iBLG from the circulation was more rapid than iBSA and iBGG. The radioactivity in mammary tissue associated with BSA and BGG was greater than 70% immunoprecipitable throughout the 4-h test interval; 125I-OVA and 125I-BLG were less than 12% precipitable 1 and 4 h after injection. In milk obtained at 4 h, there was an approximately 10-fold greater accumulation of iBSA or iBGG than of iOVA or iBLG. These experiments demonstrate that protein antigens differ in their ability to transfer from maternal circulation into milk. The transfer into milk appeared to be in proportion to persistence of the antigens in the maternal circulation.

Topics: Animals; Antigens; Dietary Proteins; Female; Food Hypersensitivity; gamma-Globulins; Iodine Radioisotopes; Lactation; Lactoglobulins; Mice; Milk; Ovalbumin; Pregnancy; Serum Albumin, Radio-Iodinated

Twelve egg-allergic patients were selected on the basis of a clinical history of egg allergy and a positive skin-prick test (SPT) to whole egg. A study was then carried out on the ability of the patients' washed leucocytes to release histamine in the presence of whole egg, ovomucoid, ovalbumin and ovotransferrin. Histamine release (HR) from washed leucocytes was demonstrated in ten out of twelve patients, but only four out of ten released over 40% of their total histamine. Spontaneous HR ranged from 2.1-14.5% with a mean of 7.5%. There was good agreement between positive and negative HR, skin test and radioallergosorbent test (RAST) results. Concordance between the HR and skin test was found in 83%, HR and RAST in 71% and skin test and RAST in 78% of patients. However there was poor quantitative agreement between these three tests. When skin-prick tests and HR thresholds were compared, ovomucoid elicited the greatest skin test sensitivity in five out of six patients, whereas five of the same six were more sensitive to ovalbumin when judged by HR.

Topics: Adolescent; Adult; Cells, Cultured; Eggs; Female; Food Hypersensitivity; Histamine Release; Humans; Immunoglobulin E; Leukocytes; Male; Middle Aged; Ovalbumin; Ovomucin; Radioallergosorbent Test; Skin Tests

Adverse reactions characteristic of food intolerance are claimed to occur in susceptible individuals following exposure to various chemical additives used to colour, flavour or preserve food. The objective of the present study was to develop a method suitable for investigating the nature and mechanism of these reactions in an animal model. Our results demonstrate that intestinal responses, elicited either specifically following oral challenge by antigen or non-specifically by the direct action of a chemical, can be quantified by evaluating the intestinal extravasation (IEV) of intravenously administered 125I-labelled rat albumin.

Topics: Animals; Extravasation of Diagnostic and Therapeutic Materials; Food Additives; Food Coloring Agents; Food Hypersensitivity; Injections, Intraperitoneal; Injections, Intravenous; Intestines; Iodine Radioisotopes; Male; Models, Biological; Ovalbumin; Rats

Topics: Adolescent; Antibody Specificity; Child; Child, Preschool; Dermatitis, Atopic; Eggs; Enzyme-Linked Immunosorbent Assay; Food Hypersensitivity; Humans; Immunoglobulin E; Immunoglobulin G; Infant; Mites; Ovalbumin; Radioallergosorbent Test

Topics: Adult; Antibody Specificity; Child; Child, Preschool; Dermatitis, Atopic; Eggs; Enzyme-Linked Immunosorbent Assay; Female; Food Hypersensitivity; Humans; Immunoglobulin E; Immunoglobulin G; Male; Ovalbumin

Topics: Child; Child, Preschool; Female; Food Hypersensitivity; Humans; Infant; Lymphocyte Activation; Lymphocytes; Male; Ovalbumin; Serum Albumin, Bovine

Serum IgG antibodies reactive with different dietary proteins have been detected in a significant proportion of adult patients with coeliac disease, dermatitis herpetiformis and atopic eczema. Serum anti-milk antibodies were shown to be distributed predominantly between the IgG2 and IgG4 subclasses, whereas anti-gliadin antibodies in atopic eczema were predominantly of the IgG4 subclass. Furthermore, as antibodies to each of these dietary antigens in healthy adults were markedly restricted to the IgG4 subclass, their production may be part of a normal immune response to dietary proteins. There was no correlation between serum IgG4 antibody and total serum IgG4 level. In contrast, restricted IgG4 anti-gliadin antibodies were less prevalent in the serum of patients with coeliac disease and dermatitis herpetiformis, suggesting defective downstream switching of Ig heavy-chain genes in these conditions.

Topics: Celiac Disease; Dermatitis Herpetiformis; Dermatitis, Atopic; Dietary Proteins; Female; Food Hypersensitivity; Gliadin; Humans; Immunoglobulin A; Immunoglobulin E; Immunoglobulin G; Immunoglobulin Isotypes; Male; Milk Proteins; Ovalbumin

An ELISA was applied to measure IgG sub-class antibodies to cow's milk beta-lactoglobulin (BLG), alpha-lactalbumin (ALA) and alpha-casein (AC) and to hen's egg ovalbumin (OA) in the sera of nineteen adult patients with milk intolerance causing either asthma, eczema or both. Results were compared with those of forty blood donors and twenty adult patients with either asthma or eczema due to inhalant allergy. Apart from one blood donor, high titres of IgG sub-class antibodies to all three milk proteins were found only in the milk intolerance group. The most frequently detected antibody was AC-specific IgG4; being high (i.e. greater than 9.98 micrograms/ml) in eight milk intolerance cases: six with eczema, one with asthma and one with both. A variable proportion of these eight patients also had high levels of IgG1, IgG2 and IgG3 antibodies to AC and IgG1, IgG2, IgG3 and IgG4 antibodies to BLG and ALA. In contrast, IgG antibody to the egg protein, OA, was remarkably restricted to IgG4 and was present in high titres in 68.4% of milk intolerant patients, 60% of inhalant allergy patients and 30% of blood donors. However, the greater incidence of high titres of IgG4 antibody to OA, compared to AC, was due to the superior coating efficiency of OA resulting in a more sensitive assay. We conclude that some adult cases of milk intolerance, particularly those with eczema, can be diagnosed by detecting raised serum levels of IgG sub-class antibodies to milk proteins.

Topics: Adolescent; Adult; Animals; Asthma; Caseins; Cattle; Eczema; Food Hypersensitivity; Humans; Immunoglobulin G; Lactoglobulins; Middle Aged; Milk; Milk Proteins; Ovalbumin

Previous studies have suggested that, apart from IgE-mediated reactions, some of the symptoms of food allergy may be caused by IgG antibodies to food proteins. This study was carried out to see if there were any distinctive features of the IgG sub-class antibody response to dietary antigens which occurs in food allergic patients. IgG sub-class antibodies were measured using a quantitative enzyme-linked immuno-sorbent assay (ELISA) to wheat gliadin, ovalbumin and bovine casein in twenty patients who had coeliac disease and in twenty-eight egg allergic patients. These were compared with twenty-one atopic dermatitis patients who did not have food allergy and twenty-six healthy control subjects. Coeliac disease patients tended to have raised IgG antibody levels (especially IgG1) to all three antigens but these overlapped considerably with that seen in egg allergic and atopic dermatitis patients. Coeliacs who avoided gluten had anti-gliadin antibody levels which did not differ from those seen in healthy subjects but nevertheless had raised anti-ovalbumin and casein-specific antibodies. The IgG antibody was largely restricted to IgG1 and IgG4 sub-class although the relative amount of each varied with the antigen. Although gliadin-specific antibodies were mainly IgG1, ovalbumin-specific antibodies were mainly IgG4. The increased antibody levels to all three antigens in coeliacs were caused by a raised IgG1 response, IgG4 antibodies were usually normal. Egg allergic patients also had raised IgG1 but not IgG4 antibodies to ovalbumin. These data show that the response to different dietary antigens can vary with the antigen. The fact that IgG1 and not IgG4 antibodies were raised to all three antigens in patients with coeliac disease suggests that they are a secondary consequence of the disease, perhaps reflecting increased transport of antigens across a damaged gut mucosa rather than a specific immunopathological reaction. However, the observation that antibodies to gliadin, and not ovalbumin or casein, fell following gluten avoidance shows that the response to gliadin, at least, is dependent upon continued exposure to antigen.

Topics: Adult; Animals; Antibodies; Caseins; Cattle; Enzyme-Linked Immunosorbent Assay; Food Hypersensitivity; Gliadin; Humans; Hypersensitivity, Delayed; Hypersensitivity, Immediate; Immunoglobulin G; Ovalbumin

Clinical parameters of 84 egg-allergic children were recorded. The individual allergens in hen's egg white were studied by means of crossed radio-immunoelectrophoresis (CRIE). Thirteen of the proteins in the egg white were found to have given rise to IgE-antibody production in the egg-allergic children. The major allergens were identified as ovalbumin, ovomucoid and ovotransferrin. Pruritus and exacerbation of atopic dermatitis were the symptoms of egg allergy most frequently recorded. A clear association was found between egg allergy and atopic dermatitis and can be explained on basis of the relationship that seems to exist between atopic dermatitis and high levels of total IgE in serum. This relationship is discussed.

Topics: Adolescent; Allergens; Child; Child, Preschool; Conalbumin; Dermatitis, Atopic; Egg White; Food Hypersensitivity; Humans; Immunoelectrophoresis, Two-Dimensional; Immunoglobulin E; Infant; Ovalbumin; Ovomucin; Radioallergosorbent Test; Skin Tests

The IgG subclasses of human antibodies against 2 dietary antigens, ovalbumin (OA) and beta-lactoglobulin (BLG), were studied by ELISA using monoclonal anti-human IgG subclass antibodies. Under the assay conditions used, the anti-IgG subclass antibodies were subclass specific. Quantitative estimates of the subclass antibodies were obtained by reference to a 'capture' assay using F(ab')2 anti-light chain antibody as ligand and IgG myelomas as standards. The validity of these estimates was supported by antibody quantitation using the Farr assay. In healthy adults with serum anti-OA or anti-BLG antibodies, anti-OA antibodies were found mainly in the IgG1 (9/11) and IgG4 (6/11) subclasses, whereas 5 sera showed high levels of IgG2 antibodies. In contrast, the IgG subclass distribution of anti-BLG antibodies was predominantly IgG4 (10/10).

Topics: Adult; Antibodies, Anti-Idiotypic; Antibodies, Monoclonal; Antibody Specificity; Diet; Enzyme-Linked Immunosorbent Assay; Female; Food Hypersensitivity; Humans; Immunoglobulin G; Lactoglobulins; Male; Myeloma Proteins; Ovalbumin

The aim of the present study was to create clearly documented immediate-type allergy to food protein in the intestine of rats and to study some pathophysiological phenomena induced by challenge with the allergen. To achieve this, rats were sensitized with ovalbumin. A passive cutaneous anaphylaxis reaction to ovalbumin was negative in all controls and positive in all test animals when Bordetella pertussis was used as adjuvant. Sixty minutes after an intravenous injection of 125I-human serum albumin and 45 min after an ovalbumin challenge, given by gavage, the rats were sacrificed. The intestine was removed and sections taken for morphologic studies. The remainder was rinsed, opened, cut into measured segments, weighed, and the radioactivity was measured. Disaccharidases, alkaline phosphatase, and protein were estimated in homogenates of epithelium. Results in both control and test animals showed that radioactivity decreased as one moved distally along the intestine. However, radioactivity was significantly higher (p less than 0.01) in the intestine of test animals than in controls. Radioactivity in liver, kidney, spleen, and lungs was identical in test and control animals. There was significant reduction in levels of alkaline phosphatase (p varied from less than 0.05 to less than 0.001), maltase (p less than 0.05), and sucrase (p less than 0.05 to less than 0.01). Lactase activity in contrast was significantly raised (p less than 0.05). There was no change in intestinal morphology or in the intestinal mast cell count.

Topics: Alkaline Phosphatase; alpha-Glucosidases; Animals; beta-Galactosidase; Cats; Food Hypersensitivity; Hypersensitivity, Immediate; Intestines; Microvilli; Milk Proteins; Ovalbumin; Passive Cutaneous Anaphylaxis; Rats; Sucrase

The occurrence of antibodies to five dietary protein antigens in the sera from 21 healthy adults was investigated by a modified Farr assay. Antibody to ovalbumin (OA) occurred most frequently (90%) whereas only 24% had antibodies to alpha-lactalbumin (ALA). No correlation was noted between the titer of antibodies against bovine serum albumin (BSA) and OA in the single individual. The avidity constants (10(8)-10(9) l/mol) and cross-reactivities against other albumins of anti-BSA antibodies in two human sera were comparable to that of the antibodies in pooled hyperimmune rabbit antiserum. Crossed radioimmunoelectrophoresis showed serum anti-BSA and anti-OA antibodies to be predominantly of the IgG class (13/13, 10/10), occasionally of the IgA- (6/13, 1/10) and rarely of the IgM class (1/13, 0/10). Analysis by radioelectroimmunoassay (rocket immunoelectrophoresis) of the IgG subclass distribution of anti-BSA and anti-OA antibodies showed total absence of IgG3. In contrast, antibodies of the IgG4 subclass were frequently present even in sera with very low levels of total IgG4.

Topics: Adult; Antibody Affinity; Antigens; Cross Reactions; Dietary Proteins; Female; Food Hypersensitivity; Humans; Immunoglobulin Allotypes; Immunoglobulin G; Male; Middle Aged; Ovalbumin; Serum Albumin, Bovine

Topics: Adolescent; Adult; Asthma; Egg White; Food Hypersensitivity; Humans; Middle Aged; Ovalbumin

An immunisation schedule is described that reproducibly induces long latent period skin sensitising antibody (immunoglobulin) in calves by a combined oral/parenteral route. This response is only directed towards the protein component of the antigen and is boosted upon challenge injection.

Topics: Animals; Cattle; Cattle Diseases; Food Hypersensitivity; Immunization; Immunoglobulin E; Lipopolysaccharides; Ovalbumin; Passive Cutaneous Anaphylaxis

In a prospective study of 92 children with at least one atopic parent, the development of the specific antibody responses to food and inhalant allergens during the first 5 years of life were assessed. By the radioallergosorbent test egg specific IgE antibody occurred in about 30% of the children with the mean peak concentration at 12 months. By the second year the prevalence of this antibody had increased whereas the mean concentration had decreased. Milk specific IgE antibody could not be shown in any subject, including four whose skin tests yielded positive results. Food specific IgG antibody was noted by antigen binding radioimmunoassays at 3 months in most children. These responses had peaked and began to fall by the fifth year. In contrast few children had detectable IgE or IgG antibody to inhalant allergens before the first 2 years of life. Both the concentration and prevalence of specific antibody, however, increased from the second to the fifth year and was greater in children whose skin tests yielded positive results. Breast feeding was associated with an increase in the prevalence of positive results from skin tests but was not associated with detectable IgE antibody to both food proteins, a lower concentration of IgG antibody to cows' milk, and was not associated with protection against the development of disease. A high level of exposure to dust mite was associated with an increased prevalence of positive results from skin tests to dust mite and appreciably higher antibody concentration. This study indicates differences in the humoral responses to food and inhalant allergens. Environmental factors appear to influence the development of these responses.

Topics: Aging; Allergens; Environmental Exposure; Food Hypersensitivity; Humans; Hypersensitivity, Immediate; Immunoglobulin E; Immunoglobulin G; Infant Nutritional Physiological Phenomena; Infant, Newborn; Lactoglobulins; Mites; Ovalbumin; Prospective Studies; Respiratory Hypersensitivity; Risk; Secale; Skin Tests

Stimulation ( [3H]thymidine incorporation) of blood lymphocytes cultured with food proteins was evaluated in infants with food protein-induced enterocolitis and correlated with the results of oral diagnostic challenges with the same foods (soy, cow's milk, and egg white). The geometric mean stimulation index for lymphocytes from patients with positive oral soy protein challenge that were cultured with soy protein was 8.5, and for patients with positive cow's milk challenge the stimulation index was 6.0 when casein was used in the cultures. Both values are significantly different from the values obtained from patients with negative oral challenges (p less than 0.01). The enhanced lymphocyte responses were specific for the food proteins responsible for clinical symptoms. It is not clear whether these lymphocyte responses are due to systemic immunization secondary to macromolecular absorption, or to an abnormality in immune regulation such as a delay in the development of oral tolerance mechanisms. They suggest, however, that circulating lymphocytes sensitive to the food antigens that produce the clinical symptoms are frequent in infants with this discrete form of food protein hypersensitivity.

Topics: Animals; Antigens; Cattle; Dose-Response Relationship, Immunologic; Enterocolitis; Food Hypersensitivity; Glycine max; Humans; Infant; Lymphocyte Activation; Lymphocytes; Milk; Milk Proteins; Ovalbumin; Plant Proteins, Dietary; Soybean Proteins

Topics: Child, Preschool; Dermatitis, Atopic; Eggs; Food Hypersensitivity; Humans; Immunoglobulin E; Infant; Ketotifen; Ovalbumin

To evaluate the role of immunologic mechanisms in one specific syndrome of food intolerance in infants, food protein-induced enterocolitis, we measured class-specific serum antibodies to three food proteins, ovalbumin, soy, and cow milk, prior to diagnostic food challenges in 18 infants suspected to have this syndrome. Infants with positive challenge reactions to egg, soy, or cow milk had 5-10 times higher levels of IgA antibody directed against that food than did the infants with negative challenges. Levels of IgG antibody to soy and egg were also significantly higher (greater than 10-fold) in infants with positive challenge responses. There was no significant difference in levels of IgM food antibodies between the two groups. IgA anti-soy antibody levels rose in all 12 infants tested 2-10 weeks after a single soy feeding (challenge). However, IgM anti-soy antibody increased in the five infants who had a negative response to the challenge feeding and decreased in those seven with a positive response. The difference between the two groups was statistically significant (P less than 0.01). Some correlation existed (r = -0.68) between the increase in IgA anti-soy antibody and the decrease in IgM anti-soy antibody for infants with positive soy challenges. Although a pathogenic role for these antibodies is not proven, the findings suggest an altered immunologic response to ingestion of food antigens in infants with food protein-induced enterocolitis.

Topics: Antibodies; Antigens; Dietary Proteins; Enterocolitis, Pseudomembranous; Food Hypersensitivity; Glycine max; Humans; Immunoglobulin G; Immunoglobulin M; Infant; Milk Proteins; Ovalbumin

The efficacy of oral SCG in preventing food allergy symptoms in children was investigated. Ten children with cow's milk and/or egg IgE-mediated allergy were selected for this study. The subjects were challenged with the offending food before and after a seven-day pre-treatment period with oral SCG (30 mg/kg b.w. per day). Full protection was achieved in six out of eight children with cow's milk allergy and in four of the five children with egg allergy. The mode of action of SCG in the prevention of clinical manifestations of food allergy is discussed.

Topics: Administration, Oral; Adolescent; Animals; Antibody Formation; Antibody Specificity; Caseins; Child; Child, Preschool; Cromolyn Sodium; Eggs; Female; Food Hypersensitivity; Humans; Immunoglobulin E; Lactalbumin; Male; Milk; Ovalbumin; Skin Tests; Skin Window Technique

Both clinical observations and experimental data support a primary role for mast cells in the genesis of immediate reactions to foods. Of particular interest is the gastrointestinal mast cell. At its location in the bowel wall it is the first mast cell to degranulate to specific antigen, provided it has antigen specific IgE on its surface. This initial reaction may potentiate entry of further specific antigen into the general circulation, where it can travel to distant sites and degranulate additional mast cells. It is also apparent that gastrointestinal mast cells in animal models have unique properties, including a reported insensitivity to certain degranulating agents and drugs. Such features must be further examined in future attempts to select therapy aimed at preventive treatments for food allergic patients.

Topics: Animals; Anti-Inflammatory Agents; Cromolyn Sodium; Cytoplasmic Granules; Digestive System; Food Hypersensitivity; Histamine; Humans; Immunoglobulin E; Intestinal Mucosa; Mast Cells; Ovalbumin; Rats

The occurrence of proteins cross-reacting with allergens in hen's egg white was studied in turkey, duck, goose and seagull egg whites, in hen egg yolk, and in hen and chicken sera and flesh. The study was based upon quantitative immunoelectrophoretic techniques. The different egg whites were all found to contain proteins cross-reacting with most of the allergens in hen's egg white, but the degree of cross-reactivity varied considerably among the various egg whites. All egg whites contained proteins able to bind human IgE-antibody in the sera of patients with allergy to hen's egg white. Several proteins cross-reacting with allergens in hen's egg white were also detected in egg yolk and in hen and chicken sera and flesh. Clinical implications of the results are discussed.

Topics: Allergens; Animals; Birds; Conalbumin; Cross Reactions; Egg Proteins; Egg White; Food Hypersensitivity; Immunoelectrophoresis; Ovalbumin; Ovomucin; Poultry; Species Specificity

Antigen 22 (ag-22) in hen's egg white, previously shown to be one of the major allergens in the egg white, was partially purified by combining biochemical separation techniques and quantitative immunoelectrophoretic methods. The molecular weight of ag-22 was found to be approximately 78,000 using analytical ultracentrifugation and pI was determined to be 6.1, which is appropriate with the values of ovotransferrin. It was concluded that ag-22 is identical with ovotransferrin. The ability of ovotransferrin to react in the human IgE-system was demonstrated in vivo and in vitro, by means of skin prick tests and crossed radioimmunoelectrophoresis.

Topics: Allergens; Animals; Antigens; Chickens; Egg Proteins; Egg White; Female; Food Hypersensitivity; Humans; Immune Sera; Immunoelectrophoresis, Two-Dimensional; Molecular Weight; Ovalbumin; Rabbits; Skin Tests

Forty-two patients with a history of egg sensitivity were evaluated for receiving avian-grown vaccines. After giving a history and undergoing physical examination, each patient was skin-tested with egg antigens and six egg-propagated vaccines, given an oral egg challenge, and finally, when possible, given a vaccine challenge. Thirty-seven of the 42 patients (88%) were ultimately given one or more of the vaccines with no reactions or only minimal ones to both egg protein and vaccine; they had mild reactions consisting of pruritus, headache, and apprehension. Immunization was withheld from three patients who had a history of severe reactions after egg ingestion and strongly positive skin reactivity to both egg and vaccine. A history of egg intolerance should not, by itself, disqualify a patient from receiving one of these vaccines. However, a history of exquisite sensitivity to egg protein indicates that a severe vaccine reaction is likely. An intradermal skin test utilizing 0.02 ml of a 1:100 dilution of the vaccine and resulting in a wheal of greater than 5 mm was found to be the test that most reliably predicted those patients who should not receive the vaccine.

Topics: Administration, Oral; Adolescent; Adult; Child; Child, Preschool; Eggs; Female; Food Hypersensitivity; Humans; Immunization; Male; Middle Aged; Ovalbumin; Skin Tests; Viral Vaccines

Topics: Alveolitis, Extrinsic Allergic; Animals; Buffaloes; Cattle; Female; Food Hypersensitivity; Humans; Infant; Milk; Ovalbumin; Precipitins

This report is of seven-year-old girl with a lifelong history of severe eczema, intestinal features of food allergy, recurrent respiratory tract infections, chronic bilateral keratitis and mucocutaneous candidiasis. Immunological tests showed high serum IgE levels, with specific IgE antibodies to cow's milk and egg white, defective PMN chemotaxis and a marked defect in both the function and number of T-lymphocytes. On a cow's milk-free and egg-free diet the eczema subsided and the respiratory infections improved. A partial correction of the immunodeficiency was also observed. The relationships between the immune system and atopy are discussed.

Topics: Animals; Antibody Formation; Cattle; Chemotaxis, Leukocyte; Child; Eggs; Eosinophilia; Female; Food Hypersensitivity; Humans; Hypergammaglobulinemia; Immunity, Cellular; Immunoglobulin E; Milk; Ovalbumin

Topics: Adolescent; Age Factors; Animals; Antibody Specificity; Chickens; Child; Child, Preschool; Egg White; Eggs; Food Hypersensitivity; Humans; Immunoglobulin E; Immunoglobulin G; Infant; Ovalbumin; Ovomucin

Topics: Allergens; Animals; Chickens; Egg White; Electrophoresis, Agar Gel; Electrophoresis, Polyacrylamide Gel; Female; Food Hypersensitivity; Humans; Immune Sera; Immunoelectrophoresis, Two-Dimensional; Isoelectric Focusing; Muramidase; Ovalbumin; Ovomucin; Rabbits; Radioallergosorbent Test

Allergens in hen's egg white were studied in crossed radio-immunoelectrophoresis (CRIE). Specific IgE-antibodies against different proteins in the egg white were examined in sera from 70 atopic patients with clinical hypersensitivity, and RAST greater than or equal to 2 to egg white. Ovalbumin, ovomucoid and an unidentified protein, antigen 22, were classified as major allergens. Specific IgE-antibodies against 10 more proteins in hen's egg white were detected. IgE-antibodies against lysozyme could not be detected.

Topics: Adolescent; Allergens; Animals; Chickens; Child; Child, Preschool; Egg White; Female; Food Hypersensitivity; Humans; Immunoelectrophoresis; Immunoelectrophoresis, Two-Dimensional; Infant; Muramidase; Ovalbumin; Ovomucin; Rabbits; Radioallergosorbent Test; Skin Tests

The intestinal absorption of ovalbumin and beta-lactoglobulin was measured in Hooded Lister rats which had previously been made allergic to ovalbumin, and in unimmunized controls. The antigens were introduced both together and separately into closed intestinal loops. Absorption of free ovalbumin, but not beta-lactoglobulin, was reduced in rats with anti-ovalbumin antibody, demonstrating antigen-specific immune exclusion despite the presence of reaginic antibody. In contrast, the absorption of beta-lactoglobulin was enhanced by the presence of ovalbumin in rats with IgE anti-ovalbumin, but not in unimmunized controls. These results suggest that macromolecular absorption may be increased in an antigen non-specific way in food allergy.

Topics: Animals; Antigen-Antibody Complex; Antigens; Epitopes; Food Hypersensitivity; Immunoglobulin E; Intestinal Absorption; Lactoglobulins; Ovalbumin; Rats

A simple two-step method for the detection of specific antigen within immune complexes is described. The immune complexes are precipitated from serum by polyethylene glycol, dissociated by incubation in acid pH buffer and adsorbed onto the surface of polystyrene tubes. The antigen is detected by the binding of a radiolabelled affinity-purified specific antibody. The assay can detect the antigen within both antigen- and antibody-excess immune complexes of any immunoglobulin class, and can also allow semiquantitative comparison of different samples. Immune complexes containing food protein antigens after eating have been found in the serum of both normal subjects and atopic patients; the latter group showed higher mean levels of antigen-specific immune complexes. The method can be adopted for large-scale screening of clinical samples for suspected antigens if suitable antisera are available.

Topics: Adult; Allergens; Animals; Antigen-Antibody Complex; Antigens; Eczema; Eggs; Food Hypersensitivity; Humans; Lactoglobulins; Methods; Milk; Ovalbumin; Polyethylene Glycols; Polystyrenes; Serum Albumin, Bovine

A case of eggwhite allergy and a case of fish allergy, where complement-fixation by thermostable IgG-class antibody was positive at extremely high antigen dilutions are described. The antibody in eggwhite allergy was of the precipitating variety and the C-fixation phenomenon was not related to the presence or absence of IgE antibody.

Topics: Animals; Antigen-Antibody Reactions; Chemical Precipitation; Chickens; Complement Fixation Tests; Complement System Proteins; Dust; Egg White; Fishes; Food Hypersensitivity; Guinea Pigs; Humans; Immunoglobulin E; Infant; Male; Ovalbumin; Ovomucin; Rabbits; Radioallergosorbent Test

Topics: Antigen-Antibody Complex; Antigens; Carbohydrates; Dietary Proteins; Food Hypersensitivity; Humans; Immunoglobulins; Intestinal Absorption; Milk Proteins; Ovalbumin; Proteins

A two site solid phase radioimmunoassay for detection of common food antigens is described. Bovine serum albumin, beta-lactoglobulin and ovalbumin can be detected in normal human serum at levels ranging from 0.1 to > 1000 ng/ml; sensitivity is not impaired by the presence of low levels of antibodies. Thirty min to 3 h after oral intake of milk, beta-lactoglobulin could be detected in the sera of 3 normal individuals, at a concentration of 0.1-3 ng/ml. This assay should prove useful in assessing the importance of macromolecular absorption in food allergy and in other gastrointestinal diseases.

Topics: Adult; Allergens; Animals; Cattle; Food; Food Hypersensitivity; Humans; Intestinal Absorption; Lactoglobulins; Milk; Ovalbumin; Radioimmunoassay; Serum Albumin, Bovine

Topics: Antibodies; Celiac Disease; Child, Preschool; Cromolyn Sodium; Food Hypersensitivity; Humans; Immunoglobulin G; Infant; Intestinal Absorption; Malabsorption Syndromes; Ovalbumin

An animal model of food allergy has been developed in which some aspects of the allergic response could be quantified and the effects of various drugs evaluated. The change in permeability of the intestinal tract of actively sensitized rats, after oral challenge with the sensitizing antigen, was the parameter measured. Rats were sensitized by injection of egg albumin and B. pertussis vaccine to induce reaginic antibody to egg albumin. Two weeks after sensitization, 125I-labelled bovine serum albumin (125I-labelled BSA) was injected intravenously, followed by oral challenge with egg albumin. Pieces of intestinal tissue were obtained and the amount of 125I-labelled BSA determined in a gamma counter. The amount of 125I-labelled BSA in the intestinal tissue of sensitized and challenged rats regularly showed an increase of greater than 100% above values for control rats.

Topics: Anaphylaxis; Animals; Antigens; Capillary Permeability; Cromolyn Sodium; Cyproheptadine; Disease Models, Animal; Female; Food Hypersensitivity; Hemocyanins; Immunoglobulin E; Intestines; Mast Cells; Ovalbumin; Pertussis Vaccine; Rats

Topics: Age Factors; Animals; Antibodies; Chickens; Child; Child, Preschool; Female; Food Hypersensitivity; Humans; Infant; Infant, Newborn; Male; Ovalbumin

Topics: Adolescent; Animals; Child; Child, Preschool; Dietary Proteins; Food Hypersensitivity; Glycine max; Histamine Release; Humans; Infant; Leukocytes; Lymphocyte Activation; Milk; Ovalbumin; Plant Extracts; Skin Tests; Triticum

Topics: Adolescent; Animals; Caseins; Chickens; Child; Child, Preschool; Dietary Proteins; DNA; Food Hypersensitivity; Humans; Lactalbumin; Lactoglobulins; Lymphocyte Activation; Milk Proteins; Ovalbumin; Plant Extracts; Serum Albumin, Bovine; Skin Tests; Thymidine; Triticum; Tritium

Topics: Adolescent; Antibodies; Biphenyl Compounds; Child; Child, Preschool; Chromatography, DEAE-Cellulose; Diazonium Compounds; Female; Food Hypersensitivity; gamma-Globulins; Hemagglutination Tests; Humans; Immunodiffusion; Immunoelectrophoresis; Immunoglobulins; Infant; Male; Ovalbumin

Topics: Adult; Allergens; Animals; Child; Female; Food Hypersensitivity; Humans; Male; Middle Aged; Milk; Ovalbumin; Skin Tests

Topics: Anaphylaxis; Animals; Antigen-Antibody Reactions; Egg White; Escherichia coli; Female; Food Hypersensitivity; Lipopolysaccharides; Ovalbumin; Pertussis Vaccine; Polysaccharides, Bacterial; Propranolol; Rats; Zymosan

Topics: Antibodies; Egg White; Eggs; Food Hypersensitivity; Humans; Immunoelectrophoresis; Immunoglobulin G; Immunoglobulin M; Iodine Isotopes; Ovalbumin; Radioimmunoassay

Topics: Adult; Allergens; Animals; Antigens; Cytochromes; Egg White; Female; Food Hypersensitivity; Guinea Pigs; Humans; Immunoelectrophoresis; Intestinal Absorption; Male; Ovalbumin; Passive Cutaneous Anaphylaxis; Radioimmunoassay

Topics: Amniotic Fluid; Antibodies; Antigens; Female; Food Hypersensitivity; Humans; Infant, Newborn; Meconium; Ovalbumin; Passive Cutaneous Anaphylaxis; Pregnancy; Umbilical Cord

Topics: Albumins; Antibodies; Blood; Caseins; Colitis, Ulcerative; Dietary Proteins; Food Hypersensitivity; Globulins; Glutens; Hemagglutination; Hemagglutination Tests; Humans; Lactoglobulins; Milk; Ovalbumin; Statistics as Topic

Topics: Adolescent; Animals; Antibodies; Child; Child, Preschool; Egg White; Female; Food Hypersensitivity; Humans; Immunoelectrophoresis; In Vitro Techniques; Infant; Male; Ovalbumin; Rabbits