固有淋巴细胞在早期肠源性脓毒症免疫抑制中的作用机制研究  

Mechanism of Innate Lymphoid Cells in the Immunosuppression of Early Gut-origin Sepsis

作　　者：杨登元 沈晓菲 陆晓峰[1] 赵阳 夏雪峰 管文贤[1] 杜峻峰 YANG Deng-yuan;SHEN Xiao-fei;LU Xiao-feng;ZHAO Yang;XIA Xue-feng;GUAN Wen-xian;DU Jun-feng(Department of Gastrointestinal Surgery,Drum Tower School of Clinical Medicine,Nanjing Medical University,Nanjing 210008,China;Department of Gastrointestinal Oncology,Nanjing Drum Tower Hospital,Affiliated Hospital of Nanjing University Medical School,Nanjing 210008,China;Department of General Surgery,the 7th Medical Center of PLA General Hospital,Beijing 100700,China)

机构地区：[1]南京医科大学鼓楼临床医学院胃肠外科,南京210008 [2]南京大学医学院附属鼓楼医院肿瘤科,南京210008 [3]中国人民解放军总医院第七医学中心普通外科,北京100700

出　　处：《解放军医药杂志》2020年第12期59-63,共5页Medical & Pharmaceutical Journal of Chinese People’s Liberation Army

基　　金：国家自然科学基金(81870393,81571563,81970500,81802846);江苏省自然科学基金(SBK2018040809)。

摘　　要：目的探讨固有淋巴细胞(ILCs)在早期肠源性脓毒症中的变化及其对于脓毒症免疫抑制的调控机制。方法收集2019年1月—9月确诊为肠源性脓毒症患者7例及健康志愿者7例的外周血,利用流式细胞术分析ILCs的改变;进一步运用流式分选技术,获得ILCs及外周血中性粒细胞行体外共培养,分析培养后细胞组成亚群变化及中性粒细胞程序性死亡配体1(PD-L1)表达情况,培养上清液利用酶联免疫吸附试验分析γ干扰素(IFN-γ)表达情况。结果与健康志愿者比较,肠源性脓毒症患者外周血ILC2比例显著下降,而ILC1比例显著上升(P<0.05)。同时,外周血中性粒细胞PD-L1表达显著上调,并且与ILC1的比例呈正相关(R 2=0.5959)。肠源性脓毒症患者外周血ILC2在体外白介素-12刺激下能够转变为ILC1,并且共培养实验证实由ILC2转化的ILC1直接促进中性粒细胞表达PD-L1(P<0.05)。结论肠源性脓毒症患者在早期即存在ILC2/ILC1转化,并且调控中性粒细胞表达PD-L1,促进脓毒症免疫抑制的发生。Objective To explore the changes of innate lymphoid cells(ILCs)in early gut-origin sepsis and its regulatory mechanism for immunosuppression in sepsis.Methods The peripheral blood of 7 patients with gut-origin sepsis and 7 healthy volunteers from January to September 2019 were collected,and the changes of ILCs was analyzed by flow cytometry.Furthermore,flow sorting technology was used to obtain ILCs and peripheral blood neutrophils for co-culture in vitro,and the changes in cell composition and the expression of neutrophil programmed death ligand 1(PD-L1)after culture were analyzed,the culture supernatant was analyzed by enzyme-linked immunosorbent assay to analyze the expression of interferon-γ(IFN-γ).Results Compared with healthy volunteers,the proportion of ILC2 in peripheral blood of patients with gut-origin sepsis was decreased significantly,while the proportion of ILC1 was increased significantly(P<0.05).In the meantime,the expression of PD-L1 in peripheral blood neutrophils was significantly up-regulated,and it was positively correlated with the ratio of ILC1(R 2=0.5959).Peripheral blood ILC2 in patients with gut-origin sepsis could be converted into ILC1 under the stimulation of interleukin-12 in vitro,and the co-culture experiment confirmed that ILC1 transformed by ILC2 directly promoted the expression of PD-L1 in neutrophils(P<0.05).Conclusion ILC2/ILC1 transformation is present in patients with early gut-origin sepsis,which regulates the expression of PD-L1 by neutrophils,and promotes the occurrence of immunosuppression in sepsis.

关 键 词：固有淋巴细胞 肠源性脓毒症 中性粒细胞 免疫抑制 程序性死亡配体1 Γ干扰素 

分 类 号：R631[医药卫生—外科学]