TRAB结构域包含蛋白2B在人类免疫缺陷病毒复制中的作用  

Role of TRAB domain-containing protein 2B in human immunodeficiency virus replication

作　　者：王淑妹 赵丽[1] 熊鹰 张晓维 梁国新[1] 耿文清[1] WANG Shumei;ZHAO Li;XIONG Ying;ZHANG Xiaowei;LIANG Guoxin;GENG Wenqing(Department of Laboratory Medicine,The First Hospital,China Medical University,NHC Key Laboratory of AIDS Immunology,National Clinical Research Center for Laboratory Medicine,Key Laboratory of AIDS Immunology of Liaoning Province,Key Laboratory of AIDS Immunology,Chinese Academy of Medical Sciences,Shenyang 110001,China)

机构地区：[1]中国医科大学附属第一医院检验科,国家卫生健康委员会艾滋病免疫学重点实验室,国家临床医学研究中心,辽宁省艾滋病免疫学重点实验室,中国医学科学院艾滋病免疫学重点实验室,沈阳110001

出　　处：《中国医科大学学报》2020年第7期577-581,共5页Journal of China Medical University

基　　金：国家自然科学基金(81273238);国家科技重大专项(2017ZX10201101)。

摘　　要：目的探讨TRAB结构域包含蛋白2B (TRABD2B)对人类免疫缺陷病毒(HIV)复制的影响。方法实时PCR检测人原代细胞和细胞系中TRABD2B mRNA的表达水平;人TRABD2B表达质粒p TRABD2B-GFP转染293T细胞并通过荧光显微镜观察TRABD2B在293T细胞中的定位。采用小干扰RNA (siRNA)下调293T细胞中TRABD2B的表达,24 h后感染p24为10 ng的HIV-1NL4-3-Luc (VSVG)假病毒并检测胞内荧光素酶活性。在293T细胞中共转染人TRABD2B表达质粒以及HIV-1NL4-3、HIV-2st和SIVmac239病毒质粒,或共转染不同种属TRABD2B表达质粒和HIV-1NL4-3病毒质粒,48 h后收集上清感染TZM-bl细胞,检测TZM-bl细胞中的荧光素酶活性。结果人TRABD2B主要表达于293T细胞,且主要分布在293T细胞的胞膜;siRNA敲减293T细胞中内源性TRABD2B的表达能够促进HIV-1NL4-3的复制(P <0.05);人TRABD2B能够抑制HIV-1NL4-3、HIV-2st和SIVmac239的复制且其他种属的TRABD2B均能够抑制HIV-1NL4-3复制,差异有统计学意义(均P <0.05)。结论 TRABD2B能够抑制HIV-1、HIV-2和SIV的复制,为HIV抗病毒治疗提供了理论基础。Objective To study the role of TRAB domain-containing protein 2B(TRABD2B)in the replication of human immunodeficiency virus(HIV).Methods The expression of TRABD2B mRNA was analyzed in human primary cells and cell lines.The human TRABD2B expression plasmid pTRABD2B-GFP was transfected into 293T cells and the localization of TRABD2B in the cells was observed using fluorescence microscopy.Small interfering RNA(siRNA)was used to knockdown the expression of TRABD2B in 293T cells.The cells were then infected with 10 ng of p24 HIVNL4-3-Luc(VSV-G)virus particles after 24 hours,following which the intracellular luciferase activity was measured.The human TRABD2B expression plasmid and plasmids containing HIV-1NL4-3,HIV-2st,and SIVmac239 were co-transfected into 293T cells while the expression plasmids containing TRABD2B from different species and the HIV-1NL4-3 plasmid were co-transfected into 293T cells.The supernatant was collected after 48 hours and used to infect TZM-bl cells and then the luciferase activity of the TZM-bl reporter cells was measured.Results Human TRABD2B was mainly expressed and distributed in the membrane of the 293T cells.HIV-1NL4-3 replicated more effectively when endogenous TRABD2B was knocked down in 293T cells using siRNA(P<0.05).Human TRABD2B inhibited the replication of HIV-1NL4-3,HIV-2st and SIVmac239.Further,TRABD2B from different species had a strong inhibitory effect on HIV-1NL4-3 replication(all P<0.05).Conclusion TRABD2B significantly inhibits the replication of HIV-1,HIV-2 and SIV,which provides a theoretical basis for the application of TRABD2B in antiviral therapy for HIV infection.

关 键 词：人类免疫缺陷病毒 TRAB结构域包含蛋白2B 种属 抗病毒 

分 类 号：R392.1[医药卫生—免疫学]