胞外高迁移率组蛋白1对HTLV-1感染的T细胞中病毒复制的影响  

Influence of extracellular HMGB1 on the virus replication of HTLV-1 infected T cells

作　　者：王霞[1] 牛志国[1] 高彩[1] 孙启蒙[1] 王金恒[1] 宋向凤[1] 高志涛[1] 韩静贤[1] 王辉[1] 

机构地区：[1]新乡医学院免疫学研究中心,453003

出　　处：《中华微生物学和免疫学杂志》2012年第12期1039-1042,共4页Chinese Journal of Microbiology and Immunology

基　　金：国家自然科学基金（30972755,81273241）;新乡医学院重点领域招标课题（ZD2011-15）;新乡医学院研究生创新课题资助计划（YJSCX201105Z）

摘　　要：目的探讨胞外的高迁移率组蛋白1（high mobility group box 1，HMGB1）对人T淋巴细胞白血病病毒1型（human T-cell leukemi int1，HTLV-1）感染的T细胞中病毒复制的影响。方法ELISA检测HTLV．1病毒阴性T细胞系Jurkat、MOLT4细胞及HTLV-1病毒阳性的T细胞系MT2、MT4细胞培养上清中的HMGBl水平；将HTLV-1的长末端重复序列荧光素酶报告基因pHTLV-1-LTR-luc转染病毒阳性细胞MT2，随后加入终浓度为0．25、0．50、0．75μs／ml的HMGBl多克隆抗体（PcAb）及其同型对照抗体，30、100、300ns／ml的重组人HMGB1蛋白（rhHMGB1）及其对照PBS，48h后检测荧光素酶活性；在病毒阳性细胞MT2中加入终浓度为0．25斗晷／ml的HMGBlPcAb及同型对照抗体，300ng／ml的rhHMGB1及对照PBS，real．timePCR检测病毒编码基因tax、poll、pol2、p19、gag、env。结果HTLv．1病毒阴性T细胞系和HTLV．1病毒阳性细胞系培养上清中的HMGBI水平无明显差异；0．25μg／ml的HMGBlPcAb可明显抑制HTLV-1-LTR的转录活性，而300ng／ml的rhHMGB1可明显促进HTLV．1．LTR的转录活性；real．timePCR检测显示0．25恤g／ml的HMGBlPcAb可明显抑制病毒编码基因poll,po／2、gag、e删的表达，300ns／ml的rhHMGB1可明显促进pol1、pol2、gag、env的表达。结论胞外的HMGB1可促进HTLV-1病毒感染T细胞的病毒复制。Objective To investigate the influence of extracellular high mobility group box 1 （HMGB1） on viral replication in HTLV-1 infected T cells. Methods HMGB1 in culture supematants of adult T-cell leukemia virus 1 （ HTLV-1 ） virus-negative cell ： Jurkat, MOLT4 cells and HTLV-1 vires-positive cells ： MT2, MT4, was detected by ELISA; The HTLV-1 long terminal repeat reporter gene （ pHTLV-1-LTR- luc） was transfected into MT2 cells by Tfx-50-mediated transfection, and 0. 25,0. 50,0. 75 p,g/ml of HMGB1 polyclonal antibody（ HMGB1 PcAb） and its isotype control rabbit IgG antibodies, 0. 03,0. 1,0. 3 V,g/ml rhHMGB1 and its control PBS,were added into culture supernatant respectively, then luciferase activity was detected after 48 h;Similarly,0. 25 p,g/ml HMGB1 PcAb and the isotype control antibody,0. 3 p,g/ml rhH- MGB1 and the control PBS were added to the culture supernatant of MT2 cell,the viral gene,poll ,pol2 ,gag, env,etc,were performed by real-time PCR. Results Culture supernatant HMGB1 levels has no significant difference between HTLV-1 positive cells MT2 and MT4 and the other two virus-negative T cell lines;Compared with isotype control antibody group,the culture supernatant,to which is added 0. 25 μg/ml HMGB1 PcAb,can significantly inhibit the HTLV-I-LTR transcriptional activity and suppress the expressions of the viral gene poll ,pol2 ,gag,env. Compared with the control PBS,0. 3 μg/ml rhHMGB1 significantly promotes the transcriptional activity of the HTLV-1-LTR and the expressions of the viral gene poll ,pol2, gag, env. Conclusion The extracellular HMGB1 can promote viral replication of HTLV-1 infected T cells.

关 键 词：人T淋巴细胞白血病病毒1型 高迁移率组蛋白1 病毒复制 

分 类 号：R733[医药卫生—肿瘤]