ovalbumin and Diarrhea

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

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

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

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

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

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

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

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

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

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

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

γ-Aminobutyric acid (GABA) is the primary inhibitory neurotransmitter in the central nervous system, and it is produced via the enzymatic activity of glutamic acid decarboxylase (GAD). GABA generates fast biological signaling through type A receptors (GABA(A)R), an anionic channel. Intriguingly, GABA is found in the jejunum epithelium of rats. The present study intended to determine whether a functional GABA signaling system exists in the intestinal epithelium and if so whether the GABA signaling regulates intestinal epithelial functions. RT-PCR, Western blot, and immunohistochemical assays of small intestinal tissues of various species were performed to determine the expression of GABA-signaling proteins in intestinal epithelial cells. Perforated patch-clamp recording was used to measure GABA-induced transmembrane current in the small intestine epithelial cell line IEC-18. The fluid weight-to-intestine length ratio was measured in mice that were treated with GABA(A)R agonist and antagonist. The effect of GABA(A)R antagonist on allergic diarrhea was examined using a mouse model. GABA, GAD, and GABA(A)R subunits were identified in small intestine epithelial cells of mice, rats, pigs, and humans. GABA(A)R agonist induced an inward current and depolarized IEC-18. Both GABA and the GABA(A)R agonist muscimol increased intestinal fluid secretion of rats. The increased intestinal secretion was largely decreased by the GABA(A)R antagonist picrotoxin or gabazine, but not by tetrodotoxin. The expression levels of GABA-signaling proteins were increased in the intestinal epithelium of mice that were sensitized and challenged with ovalbumin (OVA). The OVA-treated mice exhibited diarrhea, which was alleviated by oral administration of gabazine or picrotoxin. An endogenous autocrine GABAergic signaling exists in the mammalian intestinal epithelium, which upregulates intestinal fluid secretion. The intestinal GABAergic signaling becomes intensified in allergic diarrhea, and inhibition of this GABA-signal system alleviates the allergic diarrhea.

Topics: Adult; Animals; Autocrine Communication; Blotting, Western; Cell Line; Diarrhea; Disease Models, Animal; GABA-A Receptor Agonists; GABA-A Receptor Antagonists; gamma-Aminobutyric Acid; Humans; Immunohistochemistry; Intestinal Mucosa; Intestinal Secretions; Intestine, Small; Male; Membrane Potentials; Mice; Mice, Inbred BALB C; Middle Aged; Ovalbumin; Patch-Clamp Techniques; Rats; Rats, Wistar; Receptors, GABA-A; Reverse Transcriptase Polymerase Chain Reaction; Signal Transduction; Swine; Young Adult

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

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

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

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

We investigated the anti-allergic effect of a new strain (Pediococcus pentosaceus Sn26, the Sn26 strain) among 59 strains isolated from Japanese fermented vegetable pickles, the Sunki pickle. The Sn26 strain increased Th1 type cytokine (IL-12 and IFN-gamma) production of Peyer's patch (PP) cells in BALB/c mice, improved the Th1/Th2 balance, and inhibited IgE production of splenocytes of ovalbumin (OVA)-induced allergic diarrheic mice. Next we demonstrated, by neutralizing IL-12 and IFN-gamma, that the Sn26 strain first induced IL-12, that IL-12 induced IFN-gamma, and that decreases in IL-4 and IgE production followed. Furthermore, oral administration of the Sn26 strain decreased serum OVA-specific IgE levels and ameliorated the appearance of diarrhea in OVA-induced allergic diarrheic mice. Based on these results, it was assumed that oral administration of the Sn26 strain ameliorated type-1 allergies through improvement of the Th1/Th2 balance and decreases in IgE production.

Topics: Animals; Diarrhea; Female; Hypersensitivity; Immunoglobulin E; Mice; Mice, Inbred BALB C; Ovalbumin; Pediococcus

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

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

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

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

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

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

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

The etiology of ulcerative colitis (UC) is to be understood. The basic pathological feature of UC is intestinal chronic inflammation. Superantigen, such as Staphylococcus enterotoxin B (SEB), is reported to compromise intestinal barrier function by increasing epithelial permeability and initiate inflammation in the intestinal mucosa. Inasmuch as anatomic position of the sinus, chronic sinusitis-derived SEB may follow the secretion and to be swallowed down to the gastrointestinal tract and induce lesions to the intestinal mucosa.. Sinus wash fluid (SWF, containing SEB) was collected from a group of patients with both chronic sinusitis (CS) and UC. A group of mice were sensitized to ovalbumin (OVA) in the presence of SWF. The sensitized mice were challenged with the specific antigen OVA. The inflammatory status of the colonic tissue was determined with histology, serology and electron microscopy. Using horseradish peroxidase (HRP) as a tracer, another group of mice was stimulated with SWF for 2 hours. The HRP activity was detected in the colonic tissue with enzymatic approaches and electron microscopy.. Epithelial hyperpermeability in colonic epithelium was induced by stimulating with SWF. The HRP activity in the colonic mucosa was almost 11 times more in the SWF treated group (3.2 +/- 0.6 microg/g tissue) than the control group (0.3 +/- 0.1 microg/g tissue). Mice were sensitized using a mixture of SWF and OVA (serum OVA-specific IgE was detected with a highest titer as 1:64). Challenge with OVA induced extensive inflammation in the colonic mucosa by showing (1) marked degranulation in mast cells (MC, 46.3 +/- 4.5%) and eosinophils (Eo, 55.7 +/- 4.2%); (2) inflammatory cell infiltration (MC = 145.2 +/- 11.4; Eo = 215.8 +/- 12.5; mononuclear cell = 258.4 +/- 15.3/mm2 tissue); (3) increased MPO activity (12.9 +/- 3.2 U/g tissue) and inflammatory scores (1.8 +/- 0.3); (4) mucosal surface ulcers; (5) edema in the lamina propria; (6) bacterial translocation and abscess formation in the subepithelial region.. Introducing Sinusitis-derived SEB-containing SWF to the gastrointestinal tract compromised colonic mucosal barrier function increasing epithelial permeability to luminal macromolecular protein in mice. The SWF facilitated colonic mucosal sensitization to luminal antigen. Multiple challenging the sensitized colonic mucosa with specific antigen OVA induced inflammation, induced a condition similar to human ulcerative colitis.

Topics: Adult; Animals; Antigens, Bacterial; Chronic Disease; Colitis, Ulcerative; Colon; Diarrhea; Disease Models, Animal; Enterotoxins; Eosinophils; Female; Horseradish Peroxidase; Humans; Immunization; Immunoglobulin E; Intestinal Mucosa; Male; Mast Cells; Mice; Middle Aged; Ovalbumin; Paranasal Sinuses; Permeability; Sinusitis; Superantigens; Therapeutic Irrigation

OVA-induced allergic diarrhea occurs as a consequence of over-expression of Th1 inhibitory IL-12p40 monomers and homodimers in the large intestine, establishing a dominant Th2-type environment. In this study, we demonstrate that intranasally administered murine IL-12p70 naked DNA expression plasmids resulted in the synthesis of corresponding cytokine in the large intestinal CD11c(+) dendritic cells, leading to the inhibition of Ag-specific Th2-type response for the prevention of allergic diarrhea and the suppression of clinical symptoms including OVA-specific IgE Ab synthesis. The nasal IL-12p70 DNA treatment proved effective even after the establishment of allergic diarrhea. These results suggest that the mucosal administration of naked IL-12p70 DNA plasmid should be considered as a possible preventive and therapeutic treatment for Th2 cell-mediated food allergic diseases in the intestinal tract.

Topics: Administration, Intranasal; Colitis; Cytokines; Diarrhea; Down-Regulation; Genetic Therapy; Green Fluorescent Proteins; Immunoglobulin A; Immunoglobulin G; Interleukin-12; Intestine, Large; Lymphoid Tissue; Nasopharynx; Ovalbumin; Plasmids; Protein Subunits; Spleen; Th2 Cells; Vaccines, DNA

Atractylodes macrocephala Koidz (AMK) is well-known as a digestive and tonic material and is widely used in traditional Korean herbal medicines. Previously, we found that protein samples obtained from the medicines could induce a preferential stimulation of type 1, rather than type 2, helper T lymphocytes (Th) immune responses in vitro. Since immune response induction is controlled by the balanced activation between Th1- and Th2-type immune responses, we tested to see whether or not the AMK protein sample could inhibit the ovalbumin (OVA)-mediated allergic diarrhea, whose induction has been known to be mediated by the Th2-type immune responses. The sample treatment markedly stimulated lymphocyte proliferation, antibody production, and cytokine secretion in vitro, showing a preferential stimulation of Th1-type immune responses. In particular, oral administration of the AMK sample suppressed the OVA-mediated allergic diarrhea in mice. The sample treatment also suppressed the OVA-mediated enhanced levels of total immunoglobulin (Ig) E, as well as OVA-specific IgE, which are closely associated with Th2 cell stimulation in mice. Furthermore, the oral treatment of the sample significantly increased gamma interferon (IFN-gamma) production by lymphocytes, isolated from spleen and large intestine of the mice, that had been systematically challenged with OVA. Consequently, the oral administration of AMK protein sample suppressed the OVA-mediated allergic diarrhea by preferential stimulation of the Th1-type immune responses.

Topics: Administration, Oral; Animals; Antibody Formation; Atractylodes; Diarrhea; Immunoglobulin E; Immunoglobulin G; Interferon-gamma; Interleukin-2; Intestine, Large; Korea; Mice; Mice, Inbred BALB C; Ovalbumin; Plant Extracts; Plant Proteins; Spleen; Th1 Cells; Th2 Cells

IL-12 consists of two disulfide-linked subunits, p40 and p35, that form functionally active heterodimers for the induction of Th1 cells. In contrast to IL-12 heterodimers, p40 monomers and homodimers possess inhibitory effects on Th1 cells leading to the creation of a Th2 environment. Although it has been shown that IL-12p40 acts as antagonist of IL-12p70 in vitro, no evidence is currently available whether IL-12p40 is functional in vivo. We now report that IL-12p40 plays an important pathological role in anintestinal allergic disease. A high expression of IL-12p40 protein was demonstrated in epithelial cells, dendritic cells, and macrophages in large but not small intestine of allergic diarrhea-induced mice. Interestingly, neutralization with anti-IL-12p40 mAbs reduced the incidence and delayed the onset of disease development. Lower levels of ovalbumin (OVA)-specific IgE Abs in serum were detected in anti-IL-12p40 mAb-treated mice than in control Ab-treated mice. The secretion of Th2 cytokines and eotaxin by the mononuclear cells isolated from the large intestine of anti-IL-12p40 mAb-treated mice was significantly decreased. Finally, the removal of the IL-12p40 gene resulted in complete inhibition of disease development. These results show that over-expression of IL-12p40 is an important contributing factor for the generation of the dominant Th2-type environment in the large intestine of mice with allergic diarrhea.

Topics: Animals; Antigens, Neoplasm; Blotting, Western; Chemokine CCL11; Chemokines, CC; Cytokines; Dendritic Cells; Diarrhea; Disease Models, Animal; Enzyme-Linked Immunosorbent Assay; Epithelial Cells; Hypersensitivity; Immunoglobulin E; Immunohistochemistry; Interleukin-12; Interleukin-12 Subunit p40; Intestine, Large; Macrophages; Mice; Mice, Knockout; Mucins; Ovalbumin; Protein Subunits; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Th2 Cells

Gastrointestinal allergic disorders represent a diverse spectrum of inflammatory diseases that are occurring with increasing incidence and severity. An essential question concerning these disorders is to determine the specific cells and mediators responsible for specific clinical manifestations. With this in mind, we developed a murine model of oral allergen-induced intestinal inflammation accompanied by strong Th2-associated humoral and cellular responses and focused on the immunopathogenesis of allergic diarrhea. Exposure of OVA/alum-sensitized mice to repeated doses of intragastric OVA induced genetically restricted, dose-dependent, acute diarrhea associated with increased intestinal permeability, eosinophilia, and mastocytosis. Mice developed limited systemic manifestations of anaphylaxis, even though they developed marked intestinal mucosal mast cell degranulation. Notably, experiments involving mast cell depletion (with anti-c-kit mAb), anti-IgE treatment, and Fc epsilon RI-deficient mice indicated a critical effector role for mast cells in mediating allergic diarrhea. Furthermore, allergic diarrhea was dependent upon synergistic signaling induced by serotonin and platelet-activating factor (PAF), but not histamine. These results demonstrate that oral allergen-induced diarrhea associated with experimental Th2 intestinal inflammation is largely mast cell, IgE, serotonin, and PAF dependent.

Topics: Allergens; Anaphylaxis; Animals; Chymases; Diarrhea; Eosinophils; Immunoglobulin E; Intestinal Mucosa; Mast Cells; Mastocytosis; Mice; Mice, Inbred BALB C; Mice, Inbred C57BL; Ovalbumin; Permeability; Platelet Activating Factor; Serine Endopeptidases; Serotonin; Th2 Cells

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

The roles of mast cells and extrinsic and vagal neural pathways in the anaphylaxis-induced alterations in motility observed at sites remote from antigen exposure were explored. Rats were sensitized to egg albumin (EA) and prepared with 1) electrodes to monitor intestinal myoelectric activity, 2) an isolated intestinal loop, and 3) either intact vagal innervation or a subdiaphragmatic vagotomy. Fasting myoelectric activity was recorded before and after challenge of the jejunum in continuity or the isolated loop with EA or BSA. Intestinal segments and the brain stems were processed for mast cell identification (intestine) or Fos immunoreactivity (brain stem). EA but not BSA challenge of the jejunum or the isolated loop induced altered motility at both sites and diarrhea. Granulated mast cells were significantly reduced at the site local to but not remote from challenge. Vagotomy did not inhibit antigen-induced alterations in motility or diarrhea. The number of Fos-immunoreactive nuclei in vagal sensory or motor nuclei was not significantly altered by vagotomy. Thus antigen challenge of sensitized animals causes mast cell degranulation only at the site of direct challenge but alters motility at sites local and remote from challenge. The remote response requires intact extrinsic but not necessarily vagal neural pathways.

Topics: Anaphylaxis; Animals; Brain Stem; Cell Degranulation; Chickens; Diarrhea; Gastrointestinal Motility; Jejunum; Mast Cells; Motor Neurons; Muscle, Smooth; Neural Pathways; Ovalbumin; Proto-Oncogene Proteins c-fos; Rats; Vagotomy; Vagus Nerve

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

Microhemagglutination procedures were used to quantify antibody titers against human erythrocytes and ovalbumin in calf sera. Calves were also monitored for the prevalence of pneumonia and diarrhea. Calves, 72 males and 82 females, were the progeny of 15 AI bulls. Blood was sampled weekly for 2 wk after primary and secondary immunizations. Antibody response peaks to both antigens occurred by 14 and 7 d postimmunization, respectively. There were significant effects of season and birth and location of rearing on antibody titers against both test antigens. Diarrhea prevalence was negatively associated with high primary response antibody titers against human erythrocytes, but no trends were observed for pneumonia prevalence and for antibody titers to ovalbumin. Paternal half-sib heritability estimates ranged from 0 to .40 +/- .32 for primary antibody responses and from 0 to .87 +/- .50 for secondary antibody responses, depending on antibody specificity, and those for average titer were higher for antiovalbumin antibody (h2 = .48 +/- .39) than for antihuman erythrocyte antibody (h2 = .31 +/- .21). Although the environmental component of the humoral immune response is substantial, heritabilities of the magnitude in this study suggest the feasibility for successful genetic manipulation of antibody response profiles of young calves, and these may contribute to enhanced disease resistance.

Topics: Animals; Antibody Formation; Cattle; Cattle Diseases; Diarrhea; Erythrocytes; Female; Humans; Immunity, Innate; Male; Ovalbumin; Pneumonia; Statistics as Topic

Four trials were conducted to characterize the consumption of creep feed by nursing pigs and the effects of creep feeding (from 10 d to weaning at 28 d) on the immune response, scouring index and subsequent performance of weanling pigs. Pigs were fed a ground 20% CP corn-soybean meal-whey diet with 1.0% chromic oxide (control, 9 litters), this diet with 2.7% ovalbumin added as a dietary antigen (ovalbumin, 14 litters), or no creep feed (unexposed, 11 litters). At weaning, pigs within a litter were fed a 20% CP corn-soybean meal diet either with or without 2.7% ovalbumin. Creep-fed litters began eating at 11 d of age and disappearance of creep feed increased linearly until weaning (P less than .01). However, based on the chronic oxide coloring of the feces, total creep feed consumption was quite variable from pig to pig (13 to 194 g) and from litter to litter (107 to 1,550 g). Preweaning daily gain was similar between creep-fed and noncreep-fed litters; larger litters generally had lower daily gains (P less than .09) and less feed disappearance per pig (P less than .02). Weekly blood sampling showed that pigs fed the antigen diet had a higher (P less than .001) antibody titer to ovalbumin at 14, 21 and 28 d of age than did pigs fed the control diet or pigs unexposed to creep feed. At 56 and 63 d of age, all pigs given an ovalbumin injection at 49 d (1 ml containing 3 mg of ovalbumin) had responded (P less than .001) to injection, with the lowest titers for pigs fed the control creep diet and the highest titers for pigs fed the ovalbumin creep diet; titers were intermediate for pigs not fed creep. Regardless of preweaning or postweaning treatment, most pigs began scouring 4 to 5 d postweaning; scouring peaked at d 10 and returned to normal after d 15. Although the magnitude of difference was small, creep-fed pigs tended to scour more than pigs not fed creep (P less than .01). Postweaning performance was not influenced by preweaning treatments.

Topics: Animal Feed; Animals; Antibody Formation; Diarrhea; Eating; Female; Hemagglutination Tests; Litter Size; Male; Ovalbumin; Random Allocation; Regression Analysis; Swine; Swine Diseases; Weaning; Weight Gain

Topics: Animal Nutritional Physiological Phenomena; Animals; Animals, Newborn; Biological Transport; Colostrum; Diarrhea; Diet; Eggs; Epithelial Cells; Epithelium; Female; Intestinal Absorption; Intestinal Mucosa; Intestine, Small; Intubation, Gastrointestinal; Lactation; Macromolecular Substances; Ovalbumin; Pinocytosis; Pregnancy; Starvation; Stress, Physiological; Swine

Topics: Adolescent; Adult; Bile; Bile Acids and Salts; Calcium; Carbon Isotopes; Cholestyramine Resin; Chromates; Diarrhea; Diet; Duodenum; Fasting; Feces; Female; Gallbladder; Humans; Intestinal Absorption; Intestine, Small; Male; Methylcellulose; Middle Aged; Nitrogen; Oils; Ovalbumin; Triglycerides