白细胞介素-35对脓毒症患者CD4+和CD8+T细胞的调控作用  

The regulatory activity of interleukin-35 to CD4+and CD8+T cells in sepsis patients

作　　者：薛彦[1] 弓清梅[1] Xue Yan;Gong Qingmei(Shanxi Provincial People’s Hospital,Taiyuan 030012,China)

机构地区：[1]山西省人民医院,太原030012

出　　处：《中华急诊医学杂志》2019年第12期1524-1528,共5页Chinese Journal of Emergency Medicine

基　　金：山西省卫生与健康委员会科研计划(2017019)。

摘　　要：目的观察脓毒症(sepsis)患者血浆白细胞介素-35(IL-35)的变化,探讨IL-35对脓毒症患者CD4^+和CD8^+T细胞的调控作用。方法采用前瞻性研究方法,选择41例脓毒症患者和19例健康对照者。于就诊本院后1 h内采血,酶联免疫吸附试验(ELISA)法检测血浆IL-35水平,流式细胞术检测CD3^+、CD4^+和CD8^+T细胞计数,分析IL-35、T细胞计数与序贯器官衰竭评分(sequential organ failure assessment,SOFA)的相关性。分选脓毒症患者外周血CD4^+与CD8^+T细胞,使用重组人IL-35刺激培养48 h,实时定量PCR法检测CD4^+T细胞中T-bet、GATA-3、FoxP3、RORγt mRNA以及CD8^+T细胞中穿溶素、颗粒酶B、FasL mRNA相对表达量;ELISA法检测CD4^+T细胞培养上清中的干扰素-γ(IFN-γ)、IL-4、IL-10、IL-17水平和CD8^+T细胞培养上清中IFN-γ和肿瘤坏死因子-α(TNF-α)水平。比较IL-35刺激前后转录因子和细胞因子的变化。结果脓毒症患者血浆IL-35水平高于对照者[(76.76±10.33)pg/mL vs(27.53±8.31)pg/mL,P<0.01],但与SOFA评分无显著相关性(r=0.172,P=0.281)。脓毒症死亡组(n=11)血浆IL-35水平高于存活组(n=30)[(83.18±6.48)pg/mL vs(74.40±10.55)pg/mL,P=0.014],对两组血浆IL-35水平进行ROC曲线分析,曲线下面积为0.770,P=0.009,预测价值较好。CD3^+、CD4^+和CD8^+T细胞计数在脓毒症和对照者之间、在死亡组和存活组之间的差异均无统计学意义(均P>0.05),但与SOFA评分呈显著负相关(P<0.01)。使用重组人IL-35刺激CD4^+T细胞后,Th1转录因子T-bet和Th17转录因子RORγt mRNA降低(P<0.01),而调节性T细胞转录因子FoxP3 mRNA升高(P<0.01),但Th2转录因子GATA-3 mRNA无显著变化(P=0.745);IL-35刺激后CD4^+T细胞分泌IFN-γ和IL-17的水平降低(P<0.05),而IL-10水平提升(P<0.01),但IL-4水平无显著变化(P=0.536)。使用重组人IL-35刺激CD8^+T细胞后,穿孔素和颗粒酶B mRNA降低(P<0.01),FasL mRNA无显著变化(P=0.795),分泌IFN-γ和TNF-α的水平亦降低(P<0.05)。结论血浆IL-35�Objective To investigate plasma interleukin-35(IL-35)expression in sepsis patients,and to assess the regulatory activity of IL-35 on CD4^+and CD8^+T cells in sepsis patients.Methods A prospective study was conducted.Forty-one sepsis patients and nineteen healthy controls were enrolled in this study.According to the survival outcome on 28 day after admission,sepsis patients were further divided into the survival group(n=30)and death group(n=11).Peripheral blood samples were collected within one hour of visit.Plasma IL-35 level was measured by enzyme linked immunosorbent assay(ELISA),while CD3^+,CD4^+,and CD8^+T cell counts were measured by flow cytometry.The correlation between IL-35 expression/T cell counts and sequential organ failure assessment(SOFA)score was also assessed.Peripheral CD4^+and CD8^+T cells from sepsis patients were purified and stimulated with recombinant human IL-35 for 48 h.mRNA relative levels of T-bet,GATA-3,FoxP3,and RORγt in CD4^+T cells and perforin,granzyme B,and FasL in CD8^+T cells were measured by real-time PCR.Interferon-γ(IFN-γ),IL-4,IL-10,IL-17,and tumor necrosis factor-α(TNF-α)levels in the cultured supernatants were measured by ELISA.Transcription factor level and cytokine expression was compared prior to and post IL-35 stimulation.Results Plasma IL-35 was significantly elevated in sepsis patients when compared with controls[(76.76±10.33)pg/mL vs(27.53±8.31)pg/mL,P<0.01],however,it did not correlate with SOFA score(r=0.172,P=0.281).Plasma IL-35 was also notably increased in the death group when compared with the survival group[(83.18±6.48)pg/mL vs(74.40±10.55)pg/mL,P=0.014].Plasma IL-35 level in the death and survival groups were analyzed by receiver operating characteristic curve with an area under curve of 0.770(P=0.009).There were no remarkable differences of CD3^+,CD4^+,and CD8^+T cell counts between sepsis patients and controls,or between the death and survival groups(all P>0.05).Moreover,T cell counts were negatively correlated with SOFA score(P<0.01).IL-35 stimula

关 键 词：脓毒症 白细胞介素-35 细胞因子 T淋巴细胞 免疫 

分 类 号：R73[医药卫生—肿瘤]