CD100对脓毒性心肌病患者T淋巴细胞功能的调控作用  

Regulation of CD100 on T lymphocytes function in septic cardiomyopathy patients

作　　者：冮洪生 彭定凤[1] 胡勇钧[1] 唐哨勇 黄晴 李松海 Gang Hongsheng;Peng Dingfeng;Hu Yongjun;Tang Shaoyong;Huang Qing;Li Songhai(Department of Cardiovascular,Wuhan Fourth Hospital/Pu’ai Hospital,Tongji Medical College,Huazhong University of Science and Technology,Wuhan 430033,China)

机构地区：[1]武汉市第四医院华中科技大学同济医学院附属普爱医院心血管内科,430033

出　　处：《中华急诊医学杂志》2020年第11期1432-1438,共7页Chinese Journal of Emergency Medicine

基　　金：武汉市卫计委面上项目一般项目(WX17C11)。

摘　　要：目的观察CD100在脓毒性心肌病患者中的表达,探讨CD100对脓毒性心肌病患者T细胞功能的调控作用。方法采用前瞻性研究方法,选择2019年1月至2019年10月在武汉市第四医院(华中科技大学同济医学院附属普爱医院)就诊的37例脓毒症患者(脓毒症组)、19例脓毒性心肌病患者(脓毒性心肌病组)和24例健康对照者(对照组)。患者就诊本院后24 h内采血,分离血清和外周血单个核细胞,酶联免疫吸附试验检测血清可溶型CD100(soluble CD100,sCD100)表达,流式细胞术检测CD4^+T细胞和CD8^+T细胞表面膜型CD100(membrane-bound CD100,mCD100)表达。分选脓毒性心肌病患者CD4^+T细胞和CD8^+T细胞,使用重组人CD100刺激培养24 h,酶联免疫吸附试验检测培养上清液中干扰素-γ(Interferon-γ,IFN-γ)、白细胞介素(interleukin,IL)-17、IL-10和肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)表达。实时定量PCR法检测CD4^+T细胞T-bet、RORγt和FoxP3 mRNA相对表达量。酶联免疫斑点试验法检测CD8^+T细胞分泌穿孔素和颗粒酶B的水平。组间比较采用单因素方差分析或LSD-t检验,刺激前后比较采用配对t检验或Wilcoxon配对检验。结果脓毒性心肌病组sCD100水平(9.59±3.02)ng/mL低于对照组(13.82±2.89)ng/mL和脓毒症组(13.66±3.74)ng/mL(P<0.01),脓毒性心肌病组mCD100^+CD4^+T细胞和mCD100^+CD8^+T细胞比例均高于对照组和脓毒症组(P<0.01)。脓毒性心肌病死亡组(n=5)血清sCD100水平低于存活组(n=14)[(7.36±1.98)ng/mL vs(10.39±2.97)ng/mL,P=0.049],死亡组mCD100^+CD4^+T细胞和mCD100^+CD8^+T细胞比例均高于对照组和脓毒症组(P<0.05)。进行ROC曲线分析,曲线下面积分别为0.814、0.907和0.935,均P<0.05,预测效能均较好。重组人CD100刺激脓毒性心肌病患者CD4^+T细胞后,IFN-γ和IL-17分泌升高(P<0.05),IL-10分泌降低(P=0.0005),Th1转录因子T-bet和Th17转录因子RORγt mRNA相对表达量升高(P<0.01),调节性T细胞转录因子FoxP3 mRNA相对Objective To investigate CD100 expression in septic cardiomyopathy patients,and to explore the regulatory activity of CD100 on T lymphocytes.Methods A prospective study was conducted.Thirty-seven sepsis patients(sepsis group),nineteen septic cardiomyopathy patients(septic cardiomyopathy group),and twenty-four healthy controls(control group),who were followed up between January and October 2019 in Wuhan Fourth Hospital/Pu’ai Hospital,Tongji Medical College,Huazhong University of Science and Technology,were enrolled in the current study.Blood was collected within 24 h after visiting the hospital,and serum and peripheral blood mononuclear cells(PBMCs)were isolated.The expression of serum soluble CD100(sCD100)was measured by enzyme-linked immunosorbent assay(ELISA).Membrane-bound CD100(mCD100)expression in CD4^+T and CD8^+T cells was measured by flow cytometry.CD4^+T cells and CD8^+T cells were purified from septic cardiomyopathy patients,and were cultured with recombinant CD100 for 24 h.Interferon-γ(IFN-γ),interleukin(IL)-17,IL-10,and tumor necrosis factor-α(TNF-α)expression in the supernatants were measured by ELISA.T-bet,RORγt,and FoxP3 mRNA relative levels in CD4^+T cells were semi-quantified by real-time PCR.Perforin and granzyme B secreted by CD8^+T cells was measured by enzyme-linked immunospot assay.One-way ANOVA was used for comparison among groups,while LSD-t test was used for comparison between the two groups.Paired t test or Wilcoxon paired test was used for comparison before and after stimulation.Results Serum sCD100 in the septic cardiomyopathy group(9.59±3.02)ng/mL was lower than that in the control group(13.82±2.89)ng/mL and sepsis group(13.66±3.74)ng/mL(P<0.01).The percentages of mCD100^+CD4^+T cells and mCD100^+CD8^+T cells in the septic cardiomyopathy group were both higher than those in the control and sepsis groups(P<0.01).Serum sCD100 in the death group(n=5)was lower than that in the survival group(n=14)[(7.36±1.98)ng/mL vs(10.39±2.97)ng/mL,P=0.049].The percentages of mCD100^+CD4^+T

关 键 词：脓毒性心肌病 CD100 T淋巴细胞 免疫调控 

分 类 号：R542.2[医药卫生—心血管疾病]