ovalbumin and Inflammatory-Bowel-Diseases

Topics: Animals; Colitis; Diphtheria Toxin; Forkhead Transcription Factors; Immunotherapy, Adoptive; Inflammation; Inflammatory Bowel Diseases; Interleukin-10; Intestines; Mice; Mice, Inbred C57BL; Mice, Knockout; Ovalbumin; T-Lymphocytes, Regulatory; Transforming Growth Factor beta1

Recent studies have reported an increased incidence of inflammatory bowel disease (IBD) in patients with pulmonary diseases. Despite clinical and epidemiological studies of the interplay between colitis and asthma, the diseases' related underlying mechanisms remain unclear. In this study, we evaluated the development of colitis in a model of allergic airway inflammation. We revealed that intratracheal chronic ovalbumin (OVA) exposure induces colitis and allergic airway inflammation. Interestingly, induction of colitis was largely regulated by Th1, rather than Th2 responses, whereas allergic airway inflammation was primarily mediated by Th2 responses. Experiments in

Topics: Animals; Colitis; Inflammatory Bowel Diseases; Interferon-gamma; Mice; Mice, Knockout; Ovalbumin; Signal Transduction; T-Box Domain Proteins; Th1 Cells; Th2 Cells

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 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

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

For inflammatory bowel disease (IBD) treatment, local delivery of molecules loaded in nanoparticles to the inflamed colon could be a promising strategy. The aim of this study was to investigate how drug-loaded polymeric nanoparticles target the site of inflammation and to analyse the influence of different colon-specific delivery strategies. Three different polymeric nanoparticles were formulated using ovalbumin (OVA) as a model drug. pH-sensitive nanoparticles were made with Eudragit(®) S100. Mucoadhesive nanoparticles were created with trimethylchitosan (TMC). A mix of polymers, PLGA, PEG-PLGA and PEG-PCL, were used to obtain a sustained drug delivery. Furthermore, ligands targeting immune cells (i.e. mannose) or the inflamed colon (i.e. a specific peptide) were grafted on the PEG chain of PCL. Interaction of nanoparticles with the intestinal epithelium was explored using Caco-2 monolayers designed to mimic an inflamed epithelium and then visualized using confocal laser microscopy. TMC nanoparticles had the highest apparent permeability for OVA in the untreated model. However, in the inflamed model, there were no difference between TMC, PLGA-based and Eudragit(®) nanoparticles. The uptake of nanoparticles in the inflamed mouse colon was assessed in a horizontal diffusion chamber. Mannose-grafted PLGA nanoparticles showed the highest accumulation of OVA in inflamed colon. Based on these results, active targeting of macrophages and dendritic cells may be a promising approach for targeting the colon in IBD.

Topics: Animals; Caco-2 Cells; Chitosan; Colitis; Cytokines; Dextran Sulfate; Drug Carriers; Ethylene Oxide; Female; Humans; Inflammatory Bowel Diseases; Intestinal Absorption; Lactic Acid; Lactones; Mannose; Mice; Mice, Inbred C57BL; Nanoparticles; Ovalbumin; Polyethylene Glycols; Polyglactin 910; Polyglycolic Acid; Polylactic Acid-Polyglycolic Acid Copolymer; Polymethacrylic Acids

Less developed countries have a low incidence of immunological diseases like inflammatory bowel disease (IBD), perhaps prevented by the high prevalence of helminth infections in their populations. In the Rag IL-10(-/-) T cell transfer model of colitis, Heligmosomoides polygyrus, an intestinal helminth, prevents and reverses intestinal inflammation. This model of colitis was used to explore the importance of innate immunity in H. polygyrus protection from IBD. Rag mice briefly exposed to H. polygyrus before reconstitution with IL-10(-/-) colitogenic T cells are protected from colitis. Exposure to H. polygyrus before introduction of IL-10(-/-) and OT2 T cells reduced the capacity of the intestinal mucosa to make IFN-gamma and IL-17 after either anti-CD3 mAb or OVA stimulation. This depressed cytokine response was evident even in the absence of colitis, suggesting that the downmodulation in proinflammatory cytokine secretion was not just secondary to improvement in intestinal inflammation. Following H. polygyrus infection, dendritic cells (DCs) from the lamina propria of Rag mice displayed decreased expression of CD80 and CD86, and heightened expression of plasmacytoid dendritic cell Ag-1 and CD40. They were also less responsive to lamina proprias, producing less IL-12p40 and IL-10. Also diminished was their capacity to present OVA to OT2 T cells. These experiments infer that H. polygyrus does not require direct interactions with T or B cells to render animals resistant to colitis. DCs have an important role in driving both murine and human IBD. Data suggest that phenotypic alternations in mucosal DC function are part of the regulatory process.

Topics: Animals; Cells, Cultured; Colitis; Dendritic Cells; Disease Models, Animal; Immunity, Innate; Inflammatory Bowel Diseases; Interleukin-10; Mice; Mice, Inbred C57BL; Mice, Knockout; Mice, Transgenic; Mucous Membrane; Nematospiroides dubius; Ovalbumin; Strongylida Infections; T-Lymphocyte Subsets

To analyze the therapeutic effect of oral tolerance and nasal tolerance singly and in combination with mucosal adjuvant on experimental colitis in rats.. Rat models were established using trinitrobenzenesulphonic acid enemas. Ovalbumin was used as inducing antigen and lipopolysaccharide as adjuvant. Colonic scores, splenic mononuclear cell proliferation, and expressions of Toll-like receptors (TLR) and regulatory T cells were determined.. Colonic scores decreased most significantly after ovalbumin and lipopolysaccharide nasal administration (P<0.05). Colonic expression of forkhead box P3 in rats after ovalbumin and lipopolysaccharide oral (P<0.05) and nasal administration (P<0.01) were both significantly higher than untreated rats. TLR2 expression on CD4(+)CD25(+) T cells decreased most significantly after ovalbumin and lipopolysaccharide nasal therapies (P<0.01). TLR4 colonic expression decreased significantly after ovalbumin and lipopolysaccharide oral administration (P<0.05) and lipopolysaccharide oral administration (P<0.05).. Although experimental colitis prevented oral tolerance, nasal tolerance was successfully induced. The therapeutic effect of nasal tolerance combined with adjuvant produced the best results. TLR downregulation and CD4(+)CD25(+) T cells upregulation were involved in mucosal tolerance.

Topics: Adjuvants, Immunologic; Administration, Intranasal; Administration, Oral; Animals; Antigens; Colitis; Desensitization, Immunologic; Disease Models, Animal; Immune Tolerance; Immunity, Mucosal; Inflammatory Bowel Diseases; Intestinal Mucosa; Lipopolysaccharides; Male; Ovalbumin; Rats; Rats, Sprague-Dawley; T-Lymphocytes, Regulatory; Toll-Like Receptors; Trinitrobenzenesulfonic Acid

To evaluate the therapeutic effect of mucosal tolerance on inflammatory bowel disease (IBD).. Forty-two SD rats underwent enema of 2, 4, 6-trinitro-benzene-sulfonic acid (TNBS) so as to establish models of experimental colitis and then divided into 7 equal groups: colitis group (without treatment), oral ovalbumin (OVA) group (undergoing gastric perfusion of OVA as inducing antigen), nasal OVA group (undergoing OVA nose dropping), oral OVA plus adjuvant group [undergoing gastric perfusion OVA with lipopolysaccharide (LPS) as adjuvant], nasal OVA plus adjuvant group, adjuvant control group, and blank control group (administrated with PBS orally). Another 6 rats were used as blank control group. All of the rats were executed 21 days after enema with their spleens taken out. Splenic lymphocytes were isolated and co-cultured with OVA for 96 h. MTT method was used to calculate the proliferation index so as to detect the mucosal tolerance. Macroscopical and histolopathological scores of colon were estimated so as to evaluate the therapeutic effects.. Typical manifestations of colitis occurred 3-4 days after enema and persisted to the day 21 after enema. The macroscopical and histolopathological scores of colon of the colitis group (mean rank were 9.5 and 9.5) were both significantly higher than those of the normal control group (mean rank were 3.5 and 3.5 both P < 0.01). The splenic lymphocyte proliferation indexes of the oral OVA plus LPS, nasal OVA, and nasal OVA plus LPS groups were 0.11 +/- 0.05, 0.12 +/- 0.07, and 0.06 +/- 0.04 respectively, all significantly lower than that of the normal control group (0.30 +/- 0.13, all P < 0.05), however, without significant differences among these 3 groups (all P > 0.05). The splenic lymphocyte proliferation indexes of the oral OVA group was 0.25 +/- 0.10, not significantly different from that of the normal control group (P > 0.05). The macroscopical scores of the oral OVA plus LPS group, nasal OVA plus LPS group, and oral LPS alone group were 3.0 +/- 1.3, 2.0 +/- 0.6, and 4.3 +/- 1.0 respectively, all significantly lower than that of the colitis group (6.3 +/- 0.8, all P < 0.05); and the histological score of the oral OVA plus LPS group, nasal OVA plus LPS group, and oral LPS alone group were 3.0 +/- 1.1, 2.7 +/- 1.0, and 4.6 +/- 1.6 respectively, all significantly lower than that of the colitis group (7.5 +/- 1.0, all P < 0.05). The colonic score of the nasal OVA plus LPS group was most significantly decreased.. Colitis interferes with the induction of oral tolerance, but not the induction of nasal tolerance. Both oral tolerance combined with adjuvant and nasal tolerance combined with adjuvant have therapeutic effects on experimental colitis. Oral adjuvant alone also has therapeutic effect on colitis, and the therapeutic effect of nasal tolerance combined with adjuvant is the best.

Topics: Adjuvants, Immunologic; Animals; Immune Tolerance; Inflammatory Bowel Diseases; Intestinal Mucosa; Lipopolysaccharides; Male; Ovalbumin; Rats; Rats, Sprague-Dawley; Trinitrobenzenesulfonic Acid

Intestinal antigen uptake is enhanced in inflammatory bowel disease. We analyzed transcellular transport routes of antigens in different compartments of normal enterocytes and atypical intestinal epithelial cells called "rapid antigen uptake into the cytosol enterocytes" (RACE cells). These cells constitute a recently described population of enterocyte-derived cells, which are increased in inflammatory bowel disease. Mucosa of freshly resected specimens were incubated with the antigens ovalbumin or horseradish peroxidase. Ultrastructural labeling patterns of differentiation-dependent proteins, the brush-border enzyme sucrase-isomaltase and the cytoskeleton proteins villin and actin, were determined in enterocytes. Apoptosis was investigated biochemically and ultrastructurally by cleavage of caspase-3. Both antigens were transported to late endosomes and to trans-Golgi vesicles of enterocytes in inflammatory bowel disease and control specimens. Quantitative evaluation revealed a significantly increased transepithelial antigen transport in both compartments of RACE relative to normal enterocytes. Labeling densities for sucrase-isomaltase, villin, and actin were decreased in RACE relative to normal enterocytes. Caspase-3 was not increased in RACE cells relative to controls. RACE cells are characterized by increased antigen transport to late endosomes and the trans-Golgi network, a disassembled cytoskeleton and lower concentrations of proteins that are markers of cell differentiation.

Topics: Actins; Adult; Antigens; Apoptosis; Carrier Proteins; Caspase 3; Caspases; Cell Differentiation; Cytoskeleton; Endosomes; Enterocytes; Female; Horseradish Peroxidase; Humans; Inflammatory Bowel Diseases; Intestinal Mucosa; Male; Microfilament Proteins; Middle Aged; Ovalbumin; Protein Transport; Sucrase-Isomaltase Complex; trans-Golgi Network

Most inflammatory disorders are becoming more prevalent, especially in Western countries. The pro-inflammatory cytokine tumor necrosis factor-alpha (TNF) plays a prominent role in many of these inflammatory disorders. We have previously shown that SPRET/Ei mice exhibit an extreme and dominant resistance to high doses of TNF. In this report, we investigate the response of heterozygous (C57BL/6xSPRET/Ei)F1 mice in different models of inflammatory diseases. Compared with C57BL/6 mice, (B x S)F1 mice are protected against TNF-induced arthritis and are partially protected against allergic asthma in an ovalbumin-induced model. However, these mice display complete susceptibility to TNF-induced inflammatory bowel disease. These results indicate that the SPRET/Ei genome harbors potent dominant antiinflammatory genes that might be relevant for the treatment of certain chronic inflammatory diseases. It is very well possible that different genes are implicated in the different models.

Topics: Animals; Arthritis; Asthma; Bronchoalveolar Lavage Fluid; Crosses, Genetic; Disease Models, Animal; Female; Genetic Predisposition to Disease; Genome; Heterozygote; Inflammation; Inflammatory Bowel Diseases; Joints; Male; Mice; Mice, Inbred C57BL; Ovalbumin; Tumor Necrosis Factor-alpha

The active form of vitamin D (1,25D3) suppressed the development of animal models of human autoimmune diseases including experimental inflammatory bowel disease (IBD). The vitamin D receptor (VDR) is required for all known biologic effects of vitamin D. Here we show that VDR deficiency (knockout, KO) resulted in severe inflammation of the gastrointestinal tract in two different experimental models of IBD. In the CD45RB transfer model of IBD, CD4+/CD45RBhigh T cells from VDR KO mice induced more severe colitis than wild-type CD4+/CD45RBhigh T cells. The second model of IBD used was the spontaneous colitis that develops in IL-10 KO mice. VDR/IL-10 double KO mice developed accelerated IBD and 100% mortality by 8 wk of age. At 8 wk of age, all of the VDR and IL-10 single KO mice were healthy. Rectal bleeding was observed in every VDR/IL-10 KO mouse. Splenocytes from the VDR/IL-10 double KO mice cells transferred IBD symptoms. The severe IBD in VDR/IL-10 double KO mice is a result of the immune system and not a result of altered calcium homeostasis, or gastrointestinal tract function. The data establishes an essential role for VDR signaling in the regulation of inflammation in the gastrointestinal tract.

Topics: Animals; Cell Division; Cells, Cultured; Diet; Disease Models, Animal; Female; Genotype; Inflammatory Bowel Diseases; Lymphocytes; Male; Mice; Mice, Inbred C57BL; Mice, Knockout; Ovalbumin; Receptors, Calcitriol; Schistosomiasis mansoni

Development of topically active glucocorticosteroids with minimal systemic effects is paramount in improving therapy in inflammatory bowel disease. Our experimental model in the rat has proved useful for assessing topical versus systemic anti-inflammatory potency of glucocorticosteroids on the inflamed gut.. Experiments were performed on allergen-sensitized perfused rat ileum in vivo. Mucosal exudation of plasma, induced by local allergen perfusion, was measured as the appearance of circulating 125I-labelled albumin in the gut lumen. Experiments compared the anti-exudative effects of oral budesonide (0.1 mg/kg) with oral prednisolone (1, 3.3 or 10 mg/kg) and saline, given by oral gavage 24 h prior to allergen challenge, and of topical budesonide (3 x 10(-5) mol/L) with saline, administered in the perfusate 4 h prior to allergen challenge. Systemic glucocorticosteroid activity was assessed by weighing thymus glands after sacrifice.. Allergen-induced plasma exudation was significantly reduced by oral budesonide, oral prednisolone (dose-dependently) and topically applied budesonide; topical budesonide was effective within 4 h. While prednisolone significantly reduced the relative thymus weight at both 3.3 and 10 mg/kg, budesonide given orally, 0.1 mg/kg, or topically, 3 x 10(-5) mol/L, had no significant effect.. Budesonide, administered orally or topically, shows higher selectivity for the gut mucosa than prednisolone and produces local anti-inflammatory responses comparable to prednisolone, without the accompanying systemic effects.

Topics: Administration, Oral; Administration, Topical; Animals; Anti-Inflammatory Agents; Budesonide; Disease Models, Animal; Drug Evaluation, Preclinical; Inflammatory Bowel Diseases; Intestinal Mucosa; Male; Ovalbumin; Prednisolone; Rats; Rats, Sprague-Dawley

Induction and maintenance of peripheral tolerance are important mechanisms to maintain the balance of the immune system. In addition to the deletion of T cells and their failure to respond in certain circumstances, active suppression mediated by T cells or T-cell factors has been proposed as a mechanism for maintaining peripheral tolerance. However, the inability to isolate and clone regulatory T cells involved in antigen-specific inhibition of immune responses has made it difficult to understand the mechanisms underlying such active suppression. Here we show that chronic activation of both human and murine CD4+ T cells in the presence of interleukin (IL)-10 gives rise to CD4+ T-cell clones with low proliferative capacity, producing high levels of IL-10, low levels of IL-2 and no IL-4. These antigen-specific T-cell clones suppress the proliferation of CD4+ T cells in response to antigen, and prevent colitis induced in SCID mice by pathogenic CD4+CD45RB(high) splenic T cells. Thus IL-10 drives the generation of a CD4+ T-cell subset, designated T regulatory cells 1 (Tr1), which suppresses antigen-specific immune responses and actively downregulates a pathological immune response in vivo.

Topics: Animals; CD4-Positive T-Lymphocytes; Cells, Cultured; Clone Cells; Colitis; Cytokines; Humans; Immune Tolerance; Immunosuppression Therapy; Inflammatory Bowel Diseases; Interleukin-10; Lymphocyte Activation; Mice; Mice, Inbred BALB C; Mice, SCID; Ovalbumin; Spleen; T-Lymphocyte Subsets

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

The effects of chronic antigen feeding on systemically sensitized rats were investigated. Findings include a reduction of water and antigen intake in egg albumin (EA), sensitized rats receiving EA in their drinking water for an 8 day period, compared to that of sensitized rats fed bovine serum albumin and of naive rats. Feeding EA to sensitized animals also induced a decrease in daily weight gain. This decline did not seem to be a consequence of a decreased food intake, but might rather reflect a decreased water consumption and an alteration of nutrient absorption in the gut. Indeed, sensitized rats fed EA exhibited a significant increase in jejunal and ileal histamine content compared to control rats, which may indicate the development of an inflammatory reaction in the small intestinal mucosa. Intestinal troubles experienced because of this inflammatory reaction might explain the reduction of antigen and water intake observed in sensitized rats.

Topics: Animals; Eating; Growth Disorders; Histamine Release; Inflammatory Bowel Diseases; Intestinal Mucosa; Male; Ovalbumin; Passive Cutaneous Anaphylaxis; Rats; Rats, Inbred Strains; Reference Values; Serum Albumin, Bovine; Weight Gain