共刺激分子超家族成员sPD-1和OX40对关节炎小鼠的脾CD4+T细胞和血清细胞因子水平的影响  

Immunomodulatory effects of costimulatory molecules sPD-1 and OX40 on splenic CD4 T cells and serum cytokines in arthritic mice

作　　者：黄艳艳[1] 林书典[1] 詹锋[1] 詹宇威 肖璐 韩叶光[2] 王茹[2] 刘莉[1] HUANG Yanyan;LIN Shudian;ZHAN Feng;ZHAN Yuwei;XIAO Lu;HAN Guang;WANG Ru;LIU Li(Department of Immunology and Rheumatology,Hainan General Hospital(Hainan Affiliated Hospital of Hainan Medical University),Haikou 570311,China;Department of Clinical Laboratory,Hainan General Hospital(Hainan Affiliated Hospital of Hainan Medical University),Haikou 570311,China)

机构地区：[1]海南省人民医院(海南医学院附属海南医院)风湿免疫科,海口570311 [2]海南省人民医院(海南医学院附属海南医院)检验科,海口570311

出　　处：《免疫学杂志》2021年第5期397-403,共7页Immunological Journal

基　　金：海南省自然科学基金(819QN347)。

摘　　要：目的探讨共刺激分子超家族成员sPD-1和OX40对类风湿性关节炎(CIA)模型小鼠CD4+T细胞和血清细胞因子水平的影响,并探讨其机制。方法用DBA/1小鼠建立CIA模型,分别免疫诱导35 d和49 d。小鼠分为9组,每组10只,包括正常对照组(NC组)、急性CIA组(A-CIA组,免疫诱导35 d)、慢性CIA组(C-CIA组,免疫诱导49 d)、sPD-1组(0.1 mg/kg,i.p.)、OX40组(15 mg/kg,i.p.)、sPD-1+OX40组(0.1 mg/kg sPD-1联合15 mg/kg OX40蛋白,i.p.)、PD-1抑制剂AUNP-12组(0.2 mg/kg,i.p.)、siOX40组(60μg OX40的siRNA注射,i.v.)、阴性对照siOX40-Ctrl(60μg,i.v.)。对CIA模型小鼠的踝关节进行H&E染色并进行病理和关节肿胀评分,ELISA检测sPD-1、OX40、IL-2、IL-5、IL-4、IL-17、INF-γ水平;流式细胞术分析PD-1和OX40阳性CD4+T细胞的水平。蛋白免疫印迹分析NF-κB信号蛋白表达。结果sPD-1和OX40在CIA小鼠血清和CD4+T细胞上的表达均增加(均P<0.01);s PD-1联合OX40注射明显促进CIA模型小鼠的细胞因子IL-2、IL-5、IL-4、IL-17、INF-γ的分泌,并上调pP65的(P<0.05),抑制sPD-1和沉默OX40对CIA小鼠的细胞因子IL-2、IL-5、IL-4、IL-17、INF-γ的分泌有显著抑制作用,并抑制小鼠滑膜组织IKKβ、p-P65的表达,上调IκBα的表达(P<0.05)。结论sPD-1和OX40协同调控关节炎小鼠的炎症水平,sPD-1和OX40信号的失衡是关节炎动物机体免疫炎症的关键调节因素。Programmed death-1(PD-1),a co-stimulatory molecule,inhibits the proliferation and activation of T cells,especially reduce the secretion of various cytokines via interacting with PD-L1.OX40,another costimulatory molecule,is related to the patient’s disease activity of rheumatoid arthritis(RA).However,the effects of PD-1 and/or OX40 in RA patients is still unclear.The purpose of this study is to investigate the effect of solubility PD-1(sPD-1)and OX40 on the levels of CD4+T cells and serum cytokines in RA model mice.CIA mice model were established.Mice were divided into 9 groups,including normal control group(NC group),acute CIA group(ACIA),chronic CIA group(C-CIA),sPD-1 group,OX40 group,sPD-1+OX40 group,PD-1 inhibitor AUNP-12 group,siOX40 group,and negative control(siOX40-Ctrl).The ankle joints of CIA model mice were H&E stained and scored for pathology and joint swelling.ELISA was used to detect the levels of sPD-1,OX40,IL-2,IL-5,IL-4,IL-17,INF-γ;flow cytometry was used to analyze the levels of PD-1 and OX40 positive CD4+T cells.Furthermore,the expression of NF-κB signaling proteins were analyzed by Western blotting.Data showed that the expressions of sPD-1 and OX40 in CD4+T cells and serum CIA mouse were increased.sPD-1 combined with OX40 injection significantly promoted the secretion of inflammatory factors IL-2,IL-5,IL-4,IL-17,INF-γin CIA model mice.Inhibiting sPD-1 or silencing OX40 could inhibit the secretion of inflammatory factors IL-2,IL-5,IL-4,IL-17 and INF-γin CIA mice,downregulate the expression of IKKβand p-P65 in synovium tissue,and up-regulate the expression of IκBα.In conclusion,sPD-1 and OX40 coordinately regulate the inflammation level of arthritis mice.The imbalance of sPD-1 and OX40 signaling is a key regulator of immune inflammation in arthritic animals.

关 键 词：协同刺激分子 程序性死亡分子1 OX40 类风湿性关节炎 CIA模型 细胞因子 

分 类 号：R392.11[医药卫生—免疫学]