CD226分子影响Treg细胞功能参与小鼠自身免疫性脑脊髓膜炎的发病  

CD226 participates in the pathogenesis of experimental autoimmune encephalomyelitis by affecting Treg function

作　　者：王宁 徐曦 姜朋涛 吕明华 胡志芳 WANG Ning;XU Xi;JIANG Pengtao;Lü Minghua;HU Zhifang(Institute of Basic Medicine,Faculty of Basic Medicine,Xi’an Medical University,Xi’an 710021,China)

机构地区：[1]西安医学院基础医学部基础医学研究所,西安710021

出　　处：《山西医科大学学报》2022年第4期442-447,共6页Journal of Shanxi Medical University

基　　金：陕西省自然科学基础研究项目(2021JQ-778);西安市未央区科技局项目(201932);陕西省科技厅自然科学基础研究计划重点项目(2019JZ-38);西安医学院博士科研启动项目(2021DOC20)。

摘　　要：目的探讨CD226分子影响Treg细胞功能参与小鼠自身免疫性脑脊髓膜炎(experimental autoimmune encephalomyelitis,EAE)发病的相关机制。方法建立野生型(wild-type,WT)小鼠EAE模型(WT EAE组)和CD226基因敲除(knockout,KO)小鼠EAE模型(KO EAE组),每日监测病情和体质量变化并进行EAE评分;采用流式细胞术和qRT-PCR检测两组小鼠脾脏Treg细胞比例变化和Foxp3 mRNA相对表达水平;磁珠分选两组小鼠脾脏Treg细胞,ELISA检测细胞培养上清转化生长因子-β(transforming growth factor-β,TGF-β)和白细胞介素-10(interleukin-10,IL-10)水平;流式细胞术检测两组小鼠Treg细胞抑制性免疫分子细胞毒性T淋巴细胞相关抗原4(cytotoxic T lymphocyte antigen 4,CTLA-4)和T细胞免疫受体(T cell immune receptor with immunoglobulin and ITIM domain,TIGIT)表达水平。结果与WT EAE组相比,KO EAE组免疫后EAE发病时间迟(P<0.05),病情评分低(P<0.05),脾脏CD4+T细胞中Treg细胞比例增高(P<0.05),Foxp3 mRNA表达升高(P<0.05),Treg细胞分泌抑制性细胞因子TGF-β和IL-10水平升高(P<0.05),CTLA-4和TIGIT水平升高(P<0.05)。结论CD226分子通过影响Treg细胞数量和功能参与小鼠EAE的发生。Objective To explore the molecular mechanism of CD226 participating in the pathogenesis of experimental autoimmune encephalomyelitis(EAE)in mice by affecting the function of Treg cells.Methods EAE models were established in wild-type mice(WT EAE group)and CD226 knockout mice(KO EAE group),and the disease scores and the body weight were monitored every day.The percentage of spleen Treg cells was detected by flow cytometry,and the expression level of Foxp3 mRNA was assessed by qRT-PCR.Treg cells were seperated from the spleens of WT EAE mice or KO EAE mice by magnetic cell sorting,and the levels of TGF-βand IL-10 in the cell culture supernatant were detected by ELISA.The expression levels of inhibitory immune molecules CTLA-4 and TIGIT in Treg cell membranes in the two groups were detected by flow cytometry.Results Compared with WT EAE group,the onset was delayed in KO EAE group(P<0.05),and the disease score was decreased(P<0.05).Compared with WT EAE group,the percentage of Treg cells in CD4+T cells was increased in KO EAE group(P<0.05),and Foxp3 mRNA expression was increased(P<0.05).Compared with WT EAE group,the levels of TGF-βand IL-10 were increased in Treg cells in WT EAE group(P<0.05),and the expression levels of CTLA-4 and TIGIT in Treg cells were increased(P<0.05).Conclusion Knockout of CD226 alleviates EAE in mice by increasing the number of Treg cells and enhancing Treg function.

关 键 词：CD226 TREG细胞 FOXP3 抑制性细胞因子 免疫抑制分子 

分 类 号：R512.3[医药卫生—内科学]