活动性结核病患者自然杀伤(NK)细胞CD160的表达及其与细胞功能的关系  

Expression of CD160 in natural killer(NK)cells from patients with active tuberculosis and its relationship with cell functions

作　　者：杨秉芬 翟斐 安红娟 曹志红 刘艳华 王若 程小星 YANG Bingfen;ZHAI Fei;AN Hongjuan;CAO Zhihong;LIU Yanhua;WANG Ruo;CHENG Xiaoxing(Beijing Key Laboratory of New Techniques of Tuberculosis Diagnosis and Treatment,Military Tuberculosis Prevention and Control Key Laboratory,Senior Department of Tuberculosis,the Eighth Medical Center of Chinese PLA General Hospital,Beijing 100091,China)

机构地区：[1]中国人民解放军总医院第八医学中心结核病医学部,结核病诊疗新技术北京市重点实验室,全军结核病预防与控制重点实验室,北京100091

出　　处：《细胞与分子免疫学杂志》2022年第10期918-924,共7页Chinese Journal of Cellular and Molecular Immunology

基　　金：重大专项课题(2017ZX10201301-007-002);解放军总医院第八医学中心重点课题(2017ZD-007)。

摘　　要：目的分析外周血单个核细胞(PBMC)中CD160 mRNA和蛋白的表达与结核病免疫的关系。方法使用荧光定量PCR检测PBMC中CD160 mRNA的表达,流式细胞术分析PBMC中主要细胞群(T细胞、B细胞、自然杀伤(NK)细胞和单核细胞)表面CD160蛋白表达情况,使用流式细胞术分析NK细胞中CD160与穿孔素(perforin)、颗粒酶B(granzyme B)、颗粒溶素(granulysin)、CD69、CD107和γ干扰素(IFN-γ)的关系。结果活动性肺结核患者PBMC中CD160 mRNA表达显著性下调,且结核分枝杆菌(MTB)阳性患者显著低于MTB阴性患者;B细胞和单核细胞表面CD160表达比例低;CD3^(+)T细胞表面CD160表达在活动性肺结核患者和正常对照者之间差异不明显;活动性肺结核患者NK细胞表面CD160表达显著低于正常对照者;NK细胞与MTB抗原体外培养可下调NK细胞表面CD160的表达;活动性肺结核患者NK细胞表面活化标志CD69表达显著低于正常对照者;CD160^(+)NK细胞的perforin、granzyme B、granulysin、CD69和CD107表达都显著高于CD160^(-)NK细胞;但是CD160^(+)NK细胞的IFN-γ表达显著低于CD160^(-)NK细胞。结论活动性肺结核患者CD160 mRNA和蛋白表达显著下调,CD160促进NK细胞与结核抗原相关的活化和脱颗粒,抑制NK细胞的IFN-γ的表达,CD160可能成为结核病诊治的新靶标。Objective To investigate the relationship between the CD160 expression and anti-tuberculosis immunity.Methods Fluorescence quantitative real-time PCR was used to detect the expression of CD160 in peripheral blood mononuclear cells(PBMCs).Flow cytometry was used to analyze the expression of CD160 on main subtypes of PBMCs,such as T cells,B cells,NK cells and monocytes.The relationship among CD160 and perforin,granzyme B,granulysin,CD69,CD107 and IFN-γ in NK cells was analyzed by flow cytometry.Results CD160 mRNAs in the PBMCs from patients with active tuberculosis was significantly down-regulated,and the levels of CD160 expression in Mycobacterium tuberculosis(MTB)-positive patients was significantly lower than in MTB-negative patients.The expression of CD160 on B cells and monocytes was lower in patients with active tuberculosis as compared with normal controls,while no significant difference was observed on CD3^(+)T cells.NK cells from patients with active tuberculosis had significantly lower CD160 expression than those from normal controls.In vitro culture with MTB antigens led to down-regulated expression of CD160 on NK cells.The activation marker CD69 on NK in patients with active tuberculosis was significantly lower than that in normal controls.The expression of perforin,granzyme B,granulysin,CD69 and CD107 in CD160^(+)NK cells was significantly higher than that of CD160^(-)NK cells.However,the expression of IFN-γ in CD160^(+)NK cells was significantly lower than that of CD160^(-)NK cells.Conclusion The mRNA and protein expression of CD160 was significantly down-regulated in patients with active tuberculosis.CD160 promotes the activation and degranulation of NK cells associated with tuberculosis antigens,but suppresses the expression of IFN-γ of NK cells.CD160 may become a new target for the diagnosis and treatment of tuberculosis.

关 键 词：肺结核 NK细胞 CD160 γ干扰素(IFN-γ) 穿孔素(perforin) 颗粒酶B(granzyme B) 颗粒溶素(granulysin) CD69 CD107 

分 类 号：R521[医药卫生—内科学] R392.12[医药卫生—临床医学]