CD8~+细胞毒性T淋巴细胞对HBV感染的巨噬细胞的杀伤作用  

Killing effect of CD8+cytotoxic T-lymphocytes on HBV-infected macrophages

作　　者：程辉 臧鲁燕 孙兆霞 魏西军 李冠冲 李海波[2] CHENG Hui;ZANG Lu-yan;SUN Zhao-xi;WEI Xi-jun;LI Guan-chong;LI Hai-bo(Department of Human Anatomy,Qilu University of Medicine,Zibo 255213,China;Central Laboratory,Weifang Peo-ple's Hospital,Weifang 261041,China)

机构地区：[1]齐鲁医药学院人体解剖学教研室,山东淄博255213 [2]潍坊市人民医院中心实验室,山东潍坊261041

出　　处：《中国病理生理杂志》2021年第12期2244-2250,共7页Chinese Journal of Pathophysiology

基　　金：山东省教育厅资助项目(No.J15LK544)。

摘　　要：目的:探讨乙型肝炎病毒(HBV)感染的巨噬细胞对CD8^(+)细胞毒性T淋巴细胞杀伤功能的抵抗作用。方法:从健康人群(n=6)和HBV感染患者(n=6)中获得的肝脏活检组织,采用免疫组化方法检测CD68和HBV核心蛋白(HBc)表达。在体外,从外周血单个核细胞(PBMCs)中分离单核细胞衍生的巨噬细胞、CD4^(+)T细胞和CD8^(+)T细胞,将巨噬细胞和CD4^(+)T细胞暴露于HBV中,并与CD8^(+)T细胞以不同的效应与靶标比率共培养。采用流式细胞术分析HBc^(+)靶细胞的消除、HBV感染巨噬细胞的初始反应和HBV包膜的内化动力学。免疫荧光分析HBV包膜与和驻留蛋白不同的细胞内区室[早期内体抗原1(EEA1)、溶酶体相关膜蛋白1(LAMP1)和唾液酸结合免疫球蛋白样凝集素1(Siglec-1)]共定位情况。结果:在来自HBV感染患者的肝脏样本中观察到HBc和CD68的共定位信号。与感染HBV的CD4^(+)T细胞相比,感染HBV的巨噬细胞显著抵抗CD8^(+)T细胞的杀伤(P<0.05)。与CD4^(+)T细胞相比,CD8^(+)T细胞对感染巨噬细胞的反应倾向于产生TNF-α(P<0.01)。相对于HBV包膜与HBc共定位,HBV包膜与Siglec-1共定位显著降低(P<0.01),但仍显著高于与EEA1或LAMP1的共定位(P<0.01)。结论:HBV感染的巨噬细胞能逃避CD8^(+)T细胞的杀伤作用,这可能与HBV包膜被传递到细胞内含病毒区室有关。AIM:To investigate the resistance of hepatitis B virus(HBV)-infected macrophages to CD8^(+)cytotoxic T-lymphocytes.METHODS:The expression of CD68 and HBV core protein(HBc)in liver biopsies from healthy people(n=6)and HBV-infected patients(n=6)was detected by immunohistochemistry.In vitro,monocyte-derived macrophages,CD4^(+)T cells and CD8^(+)T cells were isolated from peripheral blood mononuclear cells(PBMCs).Macrophages and CD4^(+)T cells were exposed to HBV and co-cultured with CD8^(+)T cells at different ratios of effector to target cells.The elimination of HBc^(+)target cells,the initial reaction of macrophages infected by HBV and the internalization kinetics of HBV envelope were analyzed by flow cytometry.Immunofluorescence was used to analyze the co-localization of HBV envelope with different intracellular compartments[early endosome antigen 1(EEA1),lysosomal-associated membrane protein 1(LAMP1)and sialic acid-binding immunoglobulin-like lectin-1(Siglec-1)].RESULTS:Co-localized signals were observed for HBc and CD68 in liver samples from HBV-infected patients.Compared with HBV-infected CD4^(+)T cells,HBVinfected macrophages were intrinsically resistant to killing by CD8^(+)T cells(P<0.05).Compared with CD4^(+)T cells,CD8^(+)T cells in response to infected macrophages were skewed toward production of TNF-α(P<0.01).Although HBV envelope co-localization with Siglec-1 was significantly lower than that with HBc(P<0.01),it was significantly greater than that with either EEA1 or LAMP1(P<0.01).CONCLUSION:The HBV-infected macrophages escape the killing effect of CD8^(+)T cells,which may be related to the delivery of HBV envelope to intracellular virus-containing compartment.

关 键 词：乙型肝炎病毒 CD8~+细胞毒性T淋巴细胞 巨噬细胞 含病毒区室 

分 类 号：R512.62[医药卫生—内科学] R363.2[医药卫生—临床医学]