Th17细胞和调节性T细胞失衡参与实验性自身免疫性脑脊髓炎的发病
|
摘要
目的探讨Th17细胞和调节性T细胞(Treg)失衡在实验性自身免疫性脑脊髓炎(EAE)发病过程中的作用。方法建立EAE小鼠模型,在EAE发病的不同时期运用流式细胞术检测小鼠脾脏Treg比例变化,用实时定量PCR(qRT-PCR)分别检测小鼠脾淋巴细胞Foxp3、视黄酸相关的孤儿受体γt(RoR-γt)以及脑组织IL-17 mRNA表达,ELISA检测小鼠外周血IL-6、转化生长因子β(TGF-β)及IL-17A含量变化。结果与佐剂对照组比较,CD4+CD25+Foxp3+Treg比例和Foxp3 mRNA的表达,在EAE发病初期及高峰期明显下降,而在慢性期明显升高(P<0.05);RoR-γt mRNA表达量在发病初期与佐剂对照组比较显著增加(P<0.05),而高峰期及慢性期与佐剂对照组比较则无显著差异(P>0.05)。EAE组小鼠外周血中TGF-β、IL-6浓度在发病初期和高峰期明显升高(P<0.05);随病情发展,IL-6浓度逐渐降低,在发病慢性期与佐剂对照组比较无明显差异(P>0.05),而TGF-β浓度在发病慢性期仍较高,与佐剂对照组比较明显升高(P<0.05)。EAE组小鼠外周血中IL-17A含量在EAE不同时期均有明显升高,且高峰期升高最明显(P<0.05)。结论 Th17细胞/Treg失衡参与了EAE发病过程。
Objective To investigate the role of Th17/Treg unbalance in the pathogenesis of experimental autoimmune encephalomyelitis( EAE). Methods EAE was modeled in mice and the number of regulatory T cells( Tregs) in spleen of EAE mice was detected by flow cytometry. The expressions of Foxp3 and RoR-γt mRNA in the spleen of EAE mice and IL-17 mRNA in the brain of EAE mice were evaluated by real-time quantitative PCR and the levels of IL-6,TGF-β and IL-17 in the serum of EAE mice were examined by ELISA. Results Compared with control group,the number of CD4+CD25+Foxp3+Tregs and the expression of Foxp3 mRNA in the spleen of EAE mice dramatically decreased in the early and peak stage of EAE( P < 0. 05),but increased in chronic stage of EAE( P < 0. 05); the RoR-γt mRNA expression from mouse spleen at the early stage of EAE was significant raised( P < 0. 05),but was not significantly different at the peak and chronic stage of EAE from that in control group( P > 0. 05). The levels of IL-6 and TGF-β in the serum of EAE group dramatically increased compared with control group( P < 0. 05). With the development of EAE,the level of IL-6 gradually decreased,and there was no statistical difference in the chronic stage of EAE compared with control group( P > 0. 05). However,the level of TGF-βwas higher than that in control group in the chronic stage of EAE( P < 0. 05). Compared with those in control group,the concentration of IL-17 A and the expression of IL-17 mRNA dramatically increased in different stages of EAE group,especially in peak stage( P < 0. 05). Conclusion Th17 /Treg unbalance may be involved in the pathogenesis of EAE.
Objective To investigate the role of Th17/Treg unbalance in the pathogenesis of experimental autoimmune encephalomyelitis( EAE). Methods EAE was modeled in mice and the number of regulatory T cells( Tregs) in spleen of EAE mice was detected by flow cytometry. The expressions of Foxp3 and RoR-γt mRNA in the spleen of EAE mice and IL-17 mRNA in the brain of EAE mice were evaluated by real-time quantitative PCR and the levels of IL-6,TGF-β and IL-17 in the serum of EAE mice were examined by ELISA. Results Compared with control group,the number of CD4+CD25+Foxp3+Tregs and the expression of Foxp3 mRNA in the spleen of EAE mice dramatically decreased in the early and peak stage of EAE( P < 0. 05),but increased in chronic stage of EAE( P < 0. 05); the RoR-γt mRNA expression from mouse spleen at the early stage of EAE was significant raised( P < 0. 05),but was not significantly different at the peak and chronic stage of EAE from that in control group( P > 0. 05). The levels of IL-6 and TGF-β in the serum of EAE group dramatically increased compared with control group( P < 0. 05). With the development of EAE,the level of IL-6 gradually decreased,and there was no statistical difference in the chronic stage of EAE compared with control group( P > 0. 05). However,the level of TGF-βwas higher than that in control group in the chronic stage of EAE( P < 0. 05). Compared with those in control group,the concentration of IL-17 A and the expression of IL-17 mRNA dramatically increased in different stages of EAE group,especially in peak stage( P < 0. 05). Conclusion Th17 /Treg unbalance may be involved in the pathogenesis of EAE.
引文
[1]Fernando V,Omura S,Sato F,et al.Regulation of an autoimmune model for multiple sclerosis in Th2-biased GATA3 transgenic mice[J].Int J Mol Sci,2014,15(2):1700-1718.
[2]Geremia A,Jewell DP.The IL-23/IL-17 pathway in inflammatory bowel diseases[J].Expert Rev Gastroenterol Hepatol,2012,6(2):223-237.
[3]Yin H,Li X,Zhang B,et al.Sirolimus ameliorates inflammatory responses by switching the regulatory T/T helper type 17 profile in murine colitis[J].Immunology,2013,139(4):494-502.
[4]李宾,吴福玲,冯学斌,等.呼吸道合胞病毒毛细支气管炎患儿外周血CD4+CD25+Treg调节性T细胞与Th17细胞功能变化及意义[J].细胞与分子免疫学杂志,2012,28(4):426-428.
[5]Geremia A,Biancheri P,Allan P,et al.Innate and adaptive immunity in inflammatory bowel disease[J].Autoimmun Rev,2014,13(1):3-10.
[6]Xiao J,Liu C,Li G,et al.PDCD5 negatively regulates autoimmunity by upregulating FOXP3+regulatory T cells and suppressing Th17 and Th1 responses[J].Autoimmun,2013,47:34-44.
[7]何方园,张红鸭,胡萌萌,等.调节性T细胞与Th17细胞相关因子在实验性自身免疫性肌炎小鼠淋巴结中的表达及意义[J].细胞与分子免疫学杂志,2014,30(4):399-402.
[8]Stromnes IM,Goverman JM.Active induction of experimental allergic encephalomyelitis[J].Nat Protoc,2006,4(1):1810-1819.
[9]侯娟.Th17/Treg失衡在EAE小鼠模型免疫学发病机制中的作用[D].福建:福建医科大学,2010.
[10]朱婵虹.EAE小鼠模型的构建以及NKT细胞和Th1/Th2细胞因子的作用研究[D].福建:福建医科大学,2010.
[11]Lan Q,Fan H,Quesniaux V,et al.Induced Foxp3+regulatory T cells:a potential new weapon to treat autoimmune and inflammatory diseases[J]?Mol Cell Biol,2012,4(1):22-28.
[12]Ohkuran N,Kitagawa Y,Sakaguchi S.Development and maintenance of regulatory T cells[J].Immunity,2013,38(3):414-423.
[13]Chen Z,Lin F,Gao Y,et al.FOXP3 and RORγt:transcriptional regulation of Treg and Th17[J].Int Immunopharmacol,2011,11(5):536-539.
[14]Curotto DE,Lafaille MA,Lafaille JJ.Natural and adaptive Foxp3+regulatory T cell:more of the same or a division of labor[J].Immunity,2009,30(5):626-635.
[15]Acharya M,Mukhopadhyay S,Paidassi H,et al.αv Intergrin expression by DCs is required for Th17 cell differentiation and development of experimental autoimmune encephalomyelitis in mice[J].J Clin Invest,2010,120(12):4445-4452.
[2]Geremia A,Jewell DP.The IL-23/IL-17 pathway in inflammatory bowel diseases[J].Expert Rev Gastroenterol Hepatol,2012,6(2):223-237.
[3]Yin H,Li X,Zhang B,et al.Sirolimus ameliorates inflammatory responses by switching the regulatory T/T helper type 17 profile in murine colitis[J].Immunology,2013,139(4):494-502.
[4]李宾,吴福玲,冯学斌,等.呼吸道合胞病毒毛细支气管炎患儿外周血CD4+CD25+Treg调节性T细胞与Th17细胞功能变化及意义[J].细胞与分子免疫学杂志,2012,28(4):426-428.
[5]Geremia A,Biancheri P,Allan P,et al.Innate and adaptive immunity in inflammatory bowel disease[J].Autoimmun Rev,2014,13(1):3-10.
[6]Xiao J,Liu C,Li G,et al.PDCD5 negatively regulates autoimmunity by upregulating FOXP3+regulatory T cells and suppressing Th17 and Th1 responses[J].Autoimmun,2013,47:34-44.
[7]何方园,张红鸭,胡萌萌,等.调节性T细胞与Th17细胞相关因子在实验性自身免疫性肌炎小鼠淋巴结中的表达及意义[J].细胞与分子免疫学杂志,2014,30(4):399-402.
[8]Stromnes IM,Goverman JM.Active induction of experimental allergic encephalomyelitis[J].Nat Protoc,2006,4(1):1810-1819.
[9]侯娟.Th17/Treg失衡在EAE小鼠模型免疫学发病机制中的作用[D].福建:福建医科大学,2010.
[10]朱婵虹.EAE小鼠模型的构建以及NKT细胞和Th1/Th2细胞因子的作用研究[D].福建:福建医科大学,2010.
[11]Lan Q,Fan H,Quesniaux V,et al.Induced Foxp3+regulatory T cells:a potential new weapon to treat autoimmune and inflammatory diseases[J]?Mol Cell Biol,2012,4(1):22-28.
[12]Ohkuran N,Kitagawa Y,Sakaguchi S.Development and maintenance of regulatory T cells[J].Immunity,2013,38(3):414-423.
[13]Chen Z,Lin F,Gao Y,et al.FOXP3 and RORγt:transcriptional regulation of Treg and Th17[J].Int Immunopharmacol,2011,11(5):536-539.
[14]Curotto DE,Lafaille MA,Lafaille JJ.Natural and adaptive Foxp3+regulatory T cell:more of the same or a division of labor[J].Immunity,2009,30(5):626-635.
[15]Acharya M,Mukhopadhyay S,Paidassi H,et al.αv Intergrin expression by DCs is required for Th17 cell differentiation and development of experimental autoimmune encephalomyelitis in mice[J].J Clin Invest,2010,120(12):4445-4452.