D bystander suppression [24,25], we monitored the DSS colitis phenotype closely. There

D bystander suppression [24,25], we monitored the DSS colitis phenotype closely. There was no indication that OVA treatment led to lessened or worsened colitis severity, as judged by the investigated clinical 1315463 in the samples as compared to DSS alone. Nor did it alter the ratios of CD4+ Foxp3- (conventional T cells)/CD4+ Foxp3+ (Tregs; Figure 5B). Moreover, the percentages of IL-17A+ and IFN+ CD4+ T cells in the mLNs and spleens after ex vivo stimulation remained unchanged after the addition of OVA to the DSS model.Increased percentages of OVA-reactive splenic CD4+ Foxp3-T cells are detected in DSS and OVA treated mice after resolution of colitisTo determine if the splenic CD4+ cells were reactive to oral antigens, CD4+ cells from the mLNs and spleens were examined one week after colitis resolution in mice that were administered OVA. Cell suspensions from mLNs and spleens were restimulated with OVA and examined for the expression of the very early activation marker CD69. As shown in Figure 6A and B, OVA-reactive T cells were found in the splenic CD4+ Foxp3+ (Treg) populations of both mice treated with DSS and OVA and the OVA-treated healthy controls. However, mice treated with DSS and OVA uniquely had significantly increased percentages of splenic, OVA-reactive CD4+ Foxp3-T cells (conventional T cells). Reactive cells were not observed in the mLNs (Figure 6C and D), and no OVA-directed cells were detected in the control mice not administered OVA in the drinking water (Figure 6A ). Taken together, these results indicate that antigen-specific CD4+ Foxp3-T cells developOral OVA does not change the CD4+ T cell subset compositionTo determine if the general CD4+ T cell reactivity and subset composition had changed as a result of OVA and DSS in theAntigen-Specific T Cell Development during ColitisFigure 4. DSS and OVA treated mice have the same clinical phenotype as mice treated with DSS alone. Mice were treated with both DSS and OVA for 6 days, and several clinical parameters were measured during disease progression and after sacrifice. A) Percent weight gain relative to starting weight was measured in each individual mouse for 14 days. B) DAI was calculated on day 6 and day 1.D bystander suppression [24,25], we monitored the DSS colitis phenotype closely. There was no indication that OVA treatment led to lessened or worsened colitis severity, as judged by the investigated clinical 1317923 parameters (Figure 4A ). The weight loss (Figure 4A) and DAI (Figure 4B) measured on day 6 and 13 were normal for the DSS colitis model despite OVA in the drinking water. The total amount of cells in the mLNs (Figure 4C) and spleens (Figure 4D) was similarly increased by DSS treatment in both OVA-treated and untreated animals during sacrifice on day 14. The acute phase protein, SAA, was found to be similarly increased in the day 14 sera from both DSS-treated groups compared to control groups (Figure 4E). Histological analyses of the colons revealed no obvious differences in gross damage or general cellular infiltration between the groups (Figure 4F and G). Both the OVA-treated DSS group and the untreated DSS group had colons with apparent cellular infiltrations, crypt damage and edema (Figure 4G). No differences were detected either clinically or microscopically as a result of adding OVA to the drinking water during DSS colitis.drinking water at one week after disease resolution, the percentage of CD4+ T cells, the ratios of CD4+ Foxp3-/ CD4+Foxp3+ T cells and percentages of IL-17A+ and IFN+ T cells were determined in spleen and mLN cells after stimulation with anti-CD3. As shown in Figure 5A, administering OVA together with DSS did not change the percentage of CD4+ found 1315463 in the samples as compared to DSS alone. Nor did it alter the ratios of CD4+ Foxp3- (conventional T cells)/CD4+ Foxp3+ (Tregs; Figure 5B). Moreover, the percentages of IL-17A+ and IFN+ CD4+ T cells in the mLNs and spleens after ex vivo stimulation remained unchanged after the addition of OVA to the DSS model.Increased percentages of OVA-reactive splenic CD4+ Foxp3-T cells are detected in DSS and OVA treated mice after resolution of colitisTo determine if the splenic CD4+ cells were reactive to oral antigens, CD4+ cells from the mLNs and spleens were examined one week after colitis resolution in mice that were administered OVA. Cell suspensions from mLNs and spleens were restimulated with OVA and examined for the expression of the very early activation marker CD69. As shown in Figure 6A and B, OVA-reactive T cells were found in the splenic CD4+ Foxp3+ (Treg) populations of both mice treated with DSS and OVA and the OVA-treated healthy controls. However, mice treated with DSS and OVA uniquely had significantly increased percentages of splenic, OVA-reactive CD4+ Foxp3-T cells (conventional T cells). Reactive cells were not observed in the mLNs (Figure 6C and D), and no OVA-directed cells were detected in the control mice not administered OVA in the drinking water (Figure 6A ). Taken together, these results indicate that antigen-specific CD4+ Foxp3-T cells developOral OVA does not change the CD4+ T cell subset compositionTo determine if the general CD4+ T cell reactivity and subset composition had changed as a result of OVA and DSS in theAntigen-Specific T Cell Development during ColitisFigure 4. DSS and OVA treated mice have the same clinical phenotype as mice treated with DSS alone. Mice were treated with both DSS and OVA for 6 days, and several clinical parameters were measured during disease progression and after sacrifice. A) Percent weight gain relative to starting weight was measured in each individual mouse for 14 days. B) DAI was calculated on day 6 and day 1.