[Show abstract][Hide abstract] ABSTRACT: Background and Objective
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.
Methods and Results
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.
PLoS ONE 10/2014; 9(10):e107492. DOI:10.1371/journal.pone.0107492 · 3.23 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Aging results in attenuation of abilities to mount appropriate immune responses. The influence of aging on CD4(+) T cell migration ability toward chemokines was investigated with young and aged mice. We found functional decline in migration ability toward CCL19 and also decreased CCR7 expression level in antigen-stimulated CD4(+) T cells from aged mice compared with those from young mice. Upon addition of retinoic acid (RA), CD4(+) T cells from aged mice showed decreased CCR9 expression level compared to young mice and the migration ability of CD4(+) T cells from aged mice toward CCL25 was attenuated compared to young mice. We also observed that the expression of RALDH2 mRNA was decreased in mesenteric lymph node dendritic cells from aged mice compared to those from young mice. These results demonstrate that attenuated migration abilities of CD4(+) T cells were observed in aged mice, which correlated with decreased chemokine receptor expression. Furthermore, the reduced production and response to RA by aging may be one of the causes of such attenuated migration abilities in the intestinal immune system.
[Show abstract][Hide abstract] ABSTRACT: Inflamm-aging indicates the chronic inflammatory state resulting from increased secretion of proinflammatory cytokines and mediators such as IL-6 in the elderly. Our principle objective was to identify cell types that were affected with aging concerning IL-6 secretion in the murine model. We compared IL-6 production in spleen cells from both young and aged mice and isolated several types of cells from spleen and investigated IL-6 mRNA expression and protein production. IL-6 protein productions in cultured stromal cells from aged mice spleen were significantly high compared to young mice upon LPS stimulation. IL-6 mRNA expression level of freshly isolated stromal cells from aged mice was high compared to young mice. Furthermore, stromal cells of aged mice highly expressed IL-6 mRNA after LPS injection in vivo. These results suggest that stromal cells play a role in producing IL-6 in aged mice and imply that they contribute to the chronic inflammatory condition in the elderly.
Mediators of Inflammation 03/2014; 2014(6):826987. DOI:10.1155/2014/826987 · 3.24 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: 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.
[Show abstract][Hide abstract] ABSTRACT: Immunoglobulin (Ig) E is a mediator of food allergic reaction; however, the mechanisms of
its production in response to an ingested antigen are not fully understood. For analysis
of IgE production, here we propose an IgE response mouse model created by injection of a
Th2 cell culture and feeding of an egg white diet. According to this manipulation, total
and ovalbumin specific IgE production were elevated in this model. We think our model
enables us to analyze IgE induction by Th2 cells in food allergy and can contribute to the
development of a treatment for food allergy.
[Show abstract][Hide abstract] ABSTRACT: Oral immunotherapy with T cell epitope peptides is a promising treatment for food
allergy. We examined the effect of oral administration of an ovalbumin T cell epitope
peptide (OVA323-339) in a TCR transgenic mouse model (OVA23-3 mice). OVA23-3 mice were fed
egg-white diet containing ovalbumin and subsequently orally administrated the OVA323-339
peptide. Cytokine measurements revealed that the IL-4 production of splenic
CD4+ T cells was significantly decreased by feeding the OVA323-339 peptide.
Our study suggested that oral administration of the OVA323-339 T cell epitope peptide was
capable of inhibiting systemic IL-4 response after elicitation of predominant Th2
[Show abstract][Hide abstract] ABSTRACT: 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.
[Show abstract][Hide abstract] ABSTRACT: 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.
[Show abstract][Hide abstract] ABSTRACT: 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.
[Show abstract][Hide abstract] ABSTRACT: We examined how dietary melibiose affected the T-helper (Th) cell responses induced by an orally fed antigen in ovalbumin (OVA)-specific T cell receptor transgenic mice (OVA 23-3). Dietary melibiose markedly decreased the Th2 type responses as shown by a significant decrease in the interleukin (IL)-4 production and T cell proliferative response induced by sensitization from the 7-d oral administration of OVA. It was additionally observed that the Th1 type responses tended to decrease. We therefore examined the effect of melibiose feeding on the induction of immunological tolerance induced by the oral administration of an antigen (oral tolerance). The Th cell responses induced in BALB/c mice by subcutaneous immunization with OVA were suppressed by the prior oral administration of OVA. Such responses in the OVA-fed and immunized mice were further diminished by dietary melibiose. These results suggest that dietary melibiose strongly affected the Th cell responses to an ingested antigen, and further demonstrate the potential of melibiose to enhance the induction of oral tolerance.
[Show abstract][Hide abstract] ABSTRACT: Clarification of the mechanisms underlying the development of food-sensitive intestinal inflammation will provide an important clue to combating food allergies.
To establish a model of intestinal inflammation caused by oral administration of antigen without additional treatments, we focused on the ovalbumin (OVA) 23-3 T-cell receptor transgenic mouse, which had been reported to have high serum antigen-specific IgE responses to the feeding of an egg white diet.
Changes in body weight of mice fed an egg white diet were monitored throughout the 28-day experimental period. After the 28-day feeding, intestinal tissues were harvested for histologic examination. Endogenous production of cytokines and histamine in the jejunum, and production of cytokines secreted by OVA-specific CD4+ T cells purified from mesenteric lymph nodes, were analyzed.
Egg white diet-fed OVA23-3 mice developed weight loss and inflammation with villous atrophy and goblet cell hyperplasia, especially in the jejunum. A further characteristic feature was evidence of weight recovery and tissue repair. Jejunal inflammation was also observed in egg white diet-fed recombination activating gene (RAG)-2-deficient OVA23-3 mice. In addition, tissue sections revealed significant infiltration of specific IgE-positive cells and IgE-positive degranulating mast cells. Higher levels of IL-4 and significant levels of histamine were detected in the tissues. In the supernatant of OVA-stimulated T cells, IL-10 levels were also markedly elevated.
We report that high-dose and continuous intake of primitive OVA alone induces enteropathy containing regions under repair in OVA23-3 mice. Antigen-specific T cells and inflammatory cells primed by T(H)2 responses play important roles in regulation of development and improvement of the disease.
Long-term antigen intake causes T(H)2-dependent and food-sensitive enteropathy followed by tissue repair.
Journal of Allergy and Clinical Immunology 06/2006; 117(5):1125-32. DOI:10.1016/j.jaci.2006.01.016 · 11.48 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: To investigate the mechanisms inducing food-sensitive intestinal inflammation, we focused on the OVA23-3 mouse, a transgenic mouse strain expressing a T cell receptor that recognizes ovalbumin (OVA). Mice administered an egg-white (EW) diet containing OVA showed a trend of loose feces and significant weight loss. Histology of the jejunum showed severe inflammation with villous atrophy. Thus, we studied the role of T cells and intestinal microflora in the development of the inflammation. Severe villous disruption was observed in sections of the jejunum from OVA23-3 mice and RAG-2 gene-deficient OVA23-3 mice fed with EW-diet. Further, a larger number of T cells was found in the lamina propria of the jejunum of EW-diet fed OVA23-3 mice, RAG-2 gene-deficient mice and germfree OVA23-3 mice compared with those of control-diet fed mice. However, severe inflammation was not detected in the jejunum of germfree OVA23-3 mice. CD4+ T cells from the MLN of EW-diet fed OVA23-3 mice showed a Th2 cytokine secretion profile. These observations have thus clarified that antigen-specific Th2 cells play important roles in the development of intestinal inflammation. Although the presence of indigenous bacteria was not essential for the inflammation, T cells could mediate a more severe inflammatory response in their presence.
[Show abstract][Hide abstract] ABSTRACT: Nivalenol (NIV) has been reported to induce hyperproduction of IgA, which is regulated by T-helper 2 cells (Th2); however, whether IgE production, which is under the regulation of Th2 cells, is induced by this compound remains largely unknown. We examined the effect of NIV on antigen-specific IgE production using ovalbumin (OVA)-specific T cell receptor alphabeta-transgenic mice. The mice produced significant amounts of total and antigen-specific IgE, IgG1, and IgA in serum when given OVA orally. Administration of NIV with OVA suppressed total IgE and OVA-specific IgE, IgG1, and IgA production significantly. Cytokine assay using splenocytes obtained from mice given the OVA plus NIV diet revealed that interleukin 4 (IL-4) production was suppressed and interleuin-2 (IL-2) production was enhanced. These results suggest that the inhibition of IL-4 production and enhancement of IL-2 production induced by NIV suppressed total and antigen-specific IgE production.
[Show abstract][Hide abstract] ABSTRACT: In an effort to clarify the etiology of milk allergy from the standpoint of allergen-specific immune reactions, we investigated the determinants of IgE, IgG4, and T cells specific for bovine αs1-casein from the same individual patients by using its synthetic peptides and cyanogen bromide–digested fragments. αs1-Casein is a major allergen in cow's milk, and its unique conformation enabled us to investigate the determinants of antibodies without consideration about missing the reactivities because of conformational changes. Nine patients were selected as subjects from among 129 milk-sensitive infants screened by ELISA to assess the anti-αs1-casein IgE levels in their sera. By using ELISA for epitope mapping, a C-terminal region of αs1-casein was identified as a common binding site for IgE from all of these patients, whereas those for anti-αs1-casein IgG4 were located in multiple regions of αs1-casein. We determined the specificities of seven αs1-casein–specific T-cell lines established from peripheral blood mononuclear cells of two of the patients. These T cells have been shown to secrete IL-4. All of the T-cell lines had different specificities to αs1-casein. However, a common amino acid residue use was found among the determinants of various T-cell lines from each patient. The results suggest that patients allergic to cow's milk have characteristic B cells recognizing a limited region of αs1-casein and secreting αs1-casein–specific IgE. These B cells may interact particularly with T cells recognizing determinants with a common structure. (J Allergy Clin Immunol 1998;101:660-71)
Journal of Allergy and Clinical Immunology 05/1998; 101(5):660-671. DOI:10.1016/S0091-6749(98)70175-7 · 11.48 Impact Factor