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 leuk 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HMGB1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、0.75 μg/ml的HMGB1多克隆抗体(PcAb)及其同型对照抗体,30、100、300 ng/ml的重组人HMGB1蛋白(rhHMGB1)及其对照PBS,48 h后检测荧光素酶活性；在病毒阳性细胞MT2中加入终浓度为0.25μg/ml的HMGB1 PcAb及同型对照抗体,300ng/ml的rhHMGB1及对照PBS,real-time PCR检测病毒编码基因tax、pol1、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HMGB1水平无明显差异；0.25μg/ml的HMGB1 PcAb可明显抑制HTLV-1-LTR的转录活性,而300 ng/ml的rhHMGB1可明显促进HTLV-1-LTR的转录活性；real-time PCR检测显示0.25 μg/ml的HMGB1 PcAb可明显抑制病毒编码基因pol1、pol2、gag、env的表达,300 ng/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.

abstractsObjective 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 supernatants of adult T-cell leukemia virus 1 (HTLV-1) virus-negative cell:Jurkat,MOLT4 cells and HTLV-1 virus-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 μg/ml of HMGB1 polyclonal antibody(HMGB1 PcAb) and its isotype control rabbit IgG antibodies,0.03,0.1,0.3 μg/ml rhHMGB1 and its control PBS,were added into culture supernatant respectively,then luciferase activity was detected after 48 h;Similarly,0.25 μg/ml HMGB1 PcAb and the isotype control antibody,0.3 μg/ml rhH-MGB1 and the control PBS were added to the culture supernatant of MT2 cell,the viral gene,pol1,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-1-LTR transcriptional activity and suppress the expressions of the viral gene pol1,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 pol1,pol2,gag,env.Conclusion The extracellular HMGB1 can promote viral replication of HTLV-1 infected T cells.