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Th17 cell responses in inflammatory bowel disease
Journal of Crohn’s and Colitis (2014) 8S1, S65 S352 Available online at www.sciencedirect.com
Poster presentations Basic Science P001 miR-301a and miR-301b are highly expressed in inflamed mucosa of inflammatory bowel disease and promotes Th1/Th17 immune responses R. Wu *, W. Wu, Z. Liu. Shanghai Tenth People’s Hospital, Tongji University, Department of Gastroenterology, Shanghai, China Background: The altered expression of microRNAs (miR) has been associated with inflammatory bowel disease (IBD). Understanding the role of miR in IBD may lead to future insights into IBD pathogenesis. In this study, microRNA-301 (miR-301) was investigated and explored the potential role in the immune-regulation of the pathogenesis of IBD. Methods: Expression of miR-301a and miR-301b in the serum, peripheral blood mononuclear cells (PBMC) and inflamed mucosa of IBD patients at different stages of diseases was detected by real-time PCR. Twelve CD patients were treated with anti-TNF mAb (infliximab) at weeks 0, 2, and 6, and miR301a and miR301b were also determined in inflamed intestinal mucosa at week 10 after initial infliximab therapy. CD4+ T cells were isolated from peripheral blood of IBD patients and stimulated with anti-CD3 + anti-CD28 in the presence of inhibitor-miRNA-301a or mimics-miRNA-301a for 48h in vitro. The transcription factors of proinflammatory cytokines in CD4+ T cells were also then detected by real-time PCR. Results: Expression of miR-301a and miR-301b was significantly increased in inflamed mucosa, serum and PBMC of CD and UC patients compared with healthy donors (P < 0.05). No statistical difference was present between UC and CD groups (P > 0.05). Expression of miRNA-301a/b was observed to be markedly decreased in inflamed mucosa 10 weeks after infliximab induction therapy compared with that before infliximab therapy (P < 0.05). Interestingly, miR-301a inhibitor was found to significantly downregulate mRNA expression of IL-17A, RORC, and TNF in IBD CD4+ T cells, while miR-301a mimics could markedly promote their expression in IBD CD4+ T cells in vitro (P < 0.05). Conclusions: miR-301a and miR-301b are found to be highly increased in inflamed mucosa, serum, and PBMC of IBD patients. miRNA-301a promotes proinflammatory cytokine production (e.g., TNF) and Th17 cell differentiation in inflamed mucosa of IBD. Therefore, miRNA-301a may become an effective therapeutic target for treatment of human IBD.
P002 miR-10a suppresses dendrite cell activation and Th1/Th17 cell responses in inflammatory bowel disease W. Wu *, C. Liu, Z. Liu. Shanghai Tenth People’s Hospital, Tongji University, Department of Gastroenterology, Shanghai, China Background: Although both innate and adaptive responses to microbiota have been implicated in maintenance of intestinal homeostasis and pathogenesis of inflammatory bowel disease (IBD), it is still largely unknown how they are regulated during intestinal inflammation. In this report, we investigated the role of microRNA-10a (miR-10a), a small, evolutionarily conserved, non-coding RNA, in regulation of innate and adaptive responses to microbiota in IBD. Methods: miR-10a expression was analyzed in the inflamed mucosa of IBD patients treated with or without anti-TNF mAb (infliximab) by qRT-PCR. Human monocyte-derived dendritic cells (DC) were transfected with miR-10a precursor and inhibitor to define its effect on DC cytokine expression. Further, the role of miR-10a in regulating IBD CD4+ T cell responses was investigated in vitro. Results: The expression of miR-10a was found to be markedly decreased, while NOD2 and IL-12/IL-23p40 were significantly increased, in the inflamed mucosa of IBD patients compared to those in healthy controls. Anti-TNF mAb treatment significantly promoted miR-10a expression, whereas markedly inhibited NOD2 and IL-12/IL-23p40 in the inflamed mucosa of Crohn’s disease (CD) patients. Commensal bacteria, TNF-a, and IFN-g could inhibit human DC miR-10a expression in vitro. miR-10a was observed to downregulate DC expression of IL-12/IL-23p40 and NOD2, in that overexpression of miR-10a inhibited human DC IL-12/IL-23p40 and NOD2 expression and production of IL-12 and IL-23. Moreover, miR-10a was found to markedly suppress IBD Th1 and Th17, but not Th2, cell responses. Conclusions: Our data indicate that miR-10a is decreased in the inflamed mucosa of IBD. Further, it downregulates mucosal inflammatory response through inhibition of IL-12/IL-23p40 and NOD2 expression, and blockade of Th1/Th17 cell responses. Thus, target therapy directed against miR-10a may represent a promising approach in the treatment of IBD. P003 Vitamin D deficiency enhances adherent invasive Escherichia coli induced barrier dysfunction and experimental colonic injury A. Assa1 *, L. Pinnell2 , J. Rautava2 , L. Vong2 , K. JohnsonHenry2 , P. Sherman2 . 1 Tel Aviv University, Gastroenterology, Petach Tikvah, Israel, 2 University of Toronto, Cell Biology, Toronto, Canada Background: Adherent-invasive Escherichia coli (AIEC) colonization, has been strongly implicated to be involved in crohn’s disease and was found to be associated with ileal mucosa of Crohn’s disease patients. The aim of this study
1873-9946/$
see front matter © 2014 European Crohn’s and Colitis Organisation. Published by Elsevier B.V. All rights reserved.
Poster presentations Basic Science P001 miR-301a and miR-301b are highly expressed in inflamed mucosa of inflammatory bowel disease and promotes Th1/Th17 immune responses R. Wu *, W. Wu, Z. Liu. Shanghai Tenth People’s Hospital, Tongji University, Department of Gastroenterology, Shanghai, China Background: The altered expression of microRNAs (miR) has been associated with inflammatory bowel disease (IBD). Understanding the role of miR in IBD may lead to future insights into IBD pathogenesis. In this study, microRNA-301 (miR-301) was investigated and explored the potential role in the immune-regulation of the pathogenesis of IBD. Methods: Expression of miR-301a and miR-301b in the serum, peripheral blood mononuclear cells (PBMC) and inflamed mucosa of IBD patients at different stages of diseases was detected by real-time PCR. Twelve CD patients were treated with anti-TNF mAb (infliximab) at weeks 0, 2, and 6, and miR301a and miR301b were also determined in inflamed intestinal mucosa at week 10 after initial infliximab therapy. CD4+ T cells were isolated from peripheral blood of IBD patients and stimulated with anti-CD3 + anti-CD28 in the presence of inhibitor-miRNA-301a or mimics-miRNA-301a for 48h in vitro. The transcription factors of proinflammatory cytokines in CD4+ T cells were also then detected by real-time PCR. Results: Expression of miR-301a and miR-301b was significantly increased in inflamed mucosa, serum and PBMC of CD and UC patients compared with healthy donors (P < 0.05). No statistical difference was present between UC and CD groups (P > 0.05). Expression of miRNA-301a/b was observed to be markedly decreased in inflamed mucosa 10 weeks after infliximab induction therapy compared with that before infliximab therapy (P < 0.05). Interestingly, miR-301a inhibitor was found to significantly downregulate mRNA expression of IL-17A, RORC, and TNF in IBD CD4+ T cells, while miR-301a mimics could markedly promote their expression in IBD CD4+ T cells in vitro (P < 0.05). Conclusions: miR-301a and miR-301b are found to be highly increased in inflamed mucosa, serum, and PBMC of IBD patients. miRNA-301a promotes proinflammatory cytokine production (e.g., TNF) and Th17 cell differentiation in inflamed mucosa of IBD. Therefore, miRNA-301a may become an effective therapeutic target for treatment of human IBD.
P002 miR-10a suppresses dendrite cell activation and Th1/Th17 cell responses in inflammatory bowel disease W. Wu *, C. Liu, Z. Liu. Shanghai Tenth People’s Hospital, Tongji University, Department of Gastroenterology, Shanghai, China Background: Although both innate and adaptive responses to microbiota have been implicated in maintenance of intestinal homeostasis and pathogenesis of inflammatory bowel disease (IBD), it is still largely unknown how they are regulated during intestinal inflammation. In this report, we investigated the role of microRNA-10a (miR-10a), a small, evolutionarily conserved, non-coding RNA, in regulation of innate and adaptive responses to microbiota in IBD. Methods: miR-10a expression was analyzed in the inflamed mucosa of IBD patients treated with or without anti-TNF mAb (infliximab) by qRT-PCR. Human monocyte-derived dendritic cells (DC) were transfected with miR-10a precursor and inhibitor to define its effect on DC cytokine expression. Further, the role of miR-10a in regulating IBD CD4+ T cell responses was investigated in vitro. Results: The expression of miR-10a was found to be markedly decreased, while NOD2 and IL-12/IL-23p40 were significantly increased, in the inflamed mucosa of IBD patients compared to those in healthy controls. Anti-TNF mAb treatment significantly promoted miR-10a expression, whereas markedly inhibited NOD2 and IL-12/IL-23p40 in the inflamed mucosa of Crohn’s disease (CD) patients. Commensal bacteria, TNF-a, and IFN-g could inhibit human DC miR-10a expression in vitro. miR-10a was observed to downregulate DC expression of IL-12/IL-23p40 and NOD2, in that overexpression of miR-10a inhibited human DC IL-12/IL-23p40 and NOD2 expression and production of IL-12 and IL-23. Moreover, miR-10a was found to markedly suppress IBD Th1 and Th17, but not Th2, cell responses. Conclusions: Our data indicate that miR-10a is decreased in the inflamed mucosa of IBD. Further, it downregulates mucosal inflammatory response through inhibition of IL-12/IL-23p40 and NOD2 expression, and blockade of Th1/Th17 cell responses. Thus, target therapy directed against miR-10a may represent a promising approach in the treatment of IBD. P003 Vitamin D deficiency enhances adherent invasive Escherichia coli induced barrier dysfunction and experimental colonic injury A. Assa1 *, L. Pinnell2 , J. Rautava2 , L. Vong2 , K. JohnsonHenry2 , P. Sherman2 . 1 Tel Aviv University, Gastroenterology, Petach Tikvah, Israel, 2 University of Toronto, Cell Biology, Toronto, Canada Background: Adherent-invasive Escherichia coli (AIEC) colonization, has been strongly implicated to be involved in crohn’s disease and was found to be associated with ileal mucosa of Crohn’s disease patients. The aim of this study
1873-9946/$
see front matter © 2014 European Crohn’s and Colitis Organisation. Published by Elsevier B.V. All rights reserved.