CD30L在类风湿关节炎患者中表达及对T细胞功能的影响  

Expression of CD30L in rheumatoid arthritis patients and its effect on T cell functions

作　　者：李伟[1] 肖春媛[1] 王佳怡[1] 陆天琳 黄新芳 韩捷[1] LI Wei;XIAO Chun-yuan;WANG Jia-yi;LU Tian-lin;HUANG Xin-fang;HAN Jie(Department of Rheumatology,East Hospital Affiliated to Tongji University,Shanghai 200120,China)

机构地区：[1]同济大学附属东方医院风湿免疫科,上海200120

出　　处：《现代免疫学》2022年第5期369-374,380,共7页Current Immunology

基　　金：浦东新区科经委(PKJ2016-Y61);同济大学中央高校基本科研(22120180467)。

摘　　要：为评估类风湿关节炎(rheumatoid arthritis, RA)患者中CD30L的表达情况,并探索其对T细胞功能的影响,ELISA测定RA患者外周血及关节液中可溶性CD30配体(soluble CD30 ligand, sCD30L)的水平;FACS检测患者外周血中多种细胞表面CD30L的表达水平,观察CD30L在活化的T细胞表面表达的变化及CD30L^(+)T细胞内细胞因子的分泌水平;羧基荧光素二醋酸盐琥珀酰亚胺酯(5,6-carboxyfluorescein diacetate, succinimidyl ester, CFSE)标记联合FACS检测重组CD30L蛋白刺激对体外培养PBMC中T细胞增殖的影响。结果表明,RA患者血清中sCD30L的水平较健康对照组显著升高(P<0.01)。RA患者外周血中CD14^(+)单核细胞、CD16^(+)中性粒细胞、CD19^(+)B细胞及CD3^(+)T细胞表面CD30L的表达水平均显著高于健康对照组,以中性粒细胞的升高最为显著(均P<0.01);CD4^(+)和CD8^(+)T细胞表面CD30L的表达均显著增加,以CD4^(+)T细胞的升高更为显著(均P<0.05)。体外抗CD3和抗CD28单抗刺激条件下RA患者外周血T细胞表面CD30L分子的表达有一定规律:24~72 h升高最为显著,伴随细胞因子IFN-γ、IL-4及IL-17分泌水平的改变。重组CD30L蛋白可促进RA患者PBMC中T细胞的增殖(P<0.05)。上述结果提示,CD30L分子在RA患者中呈高表达,可能通过多种细胞协同作用影响T细胞的增殖及细胞因子分泌,参与疾病过程。This study investigated the expression of CD30 L in rheumatoid arthritis(RA)patients and explored its impact on T cell functions.For this purpose,ELISA was used to determine the levels of soluble CD30 ligand(sCD30 L)in peripheral blood and synovial fluid of RA patients,and FACS was used to detect CD30 L expression levels on cells in peripheral blood,the expression changes of CD30 L on activated T cells,and the secretion of cytokines in CD30 L^(+)T cells.In addition,5,6-carboxyfluorescein diacetate,succinimidyl ester(CFSE)labeling combined FACS was used to test the proliferation of T cells when stimulated with recombinant CD30 L protein.The results showed that the serum sCD30 L levels in RA patients were significantly higher than those of healthy controls(P<0.01).The expression of CD30 L on CD14^(+)monocytes,CD16^(+)neutrophils,CD19^(+)B cells,and CD3^(+)T cells in peripheral blood of RA patients were all significantly higher than those of healthy controls,with the most significant change in neutrophils(all P<0.01).The expression of CD30 L increased on both CD4^(+)and CD8^(+)T cells,especially in CD4^(+)T cells(all P<0.05).Consistently,the expression of CD30 L on T cells from the RA patients peripheral blood increased after being activated by anti-CD3/CD28 monoclonal antibody.The most significant increase was observed from 24 h to 72 h accompanied with the secretion changes of cytokines IFN-γ,IL-4 and IL-17.Our results also showed that recombinant CD30 L protein promoted the proliferation of T cells in peripheral blood of RA patients(P<0.05).Altogether,the results suggest that CD30 L is highly expressed in RA patients and may influence the progression of the disease by affecting the proliferation of T cells and their interaction with other immune cells by regulating the secretion of cytokines.

关 键 词：类风湿关节炎 CD30配体 共刺激分子 T细胞 

分 类 号：R593.22[医药卫生—内科学]