中文摘要：

抗原特异T细胞受体（Tcell receptor,TCR）基因修饰T细胞已被应用于肿瘤、病毒感染过继免疫治疗研究,取得了鼓舞人心的结果,但在结核上未见报道.本研究利用结核分枝杆菌38kD抗原刺激人CD4^＋,CD8^＋T细胞,通过分析刺激前后T细胞TCR互补决定区3（complementarity determining region 3,CDR3）谱型,从CD4^＋,CD8^＋T细胞中分别筛选出38kD抗原特异的TCR Vα11,Vβ8和Vα3,Vβ8基因家族T细胞,聚合酶链式反应（polymerase chain reaction,PCR）扩增获得其α,β链全长基因并插入逆转录病毒载体,构建重组载体pMX-β8-IRES-α11-GFP（pβ8α11）和pMX-β8-IRES-α3-GFP（pβ8α3）,分别转染初始CD4^＋,CD8^＋T细胞,检测其体外抗结核抗原活性.结果显示,TCR基因修饰的CD4＋,CD8＋T细胞均表达外源性TCR,不仅能特异性识别抗原,且介导免疫效应功能,表明结核抗原特异TCR基因修饰的T细胞具有应用于多药耐药结核患者过继细胞免疫治疗的可能.

英文摘要：

Recent advances in T-cell receptor （TCR） gene modification methods have enabled researchers to design T-cells specific for target antigens, e.g., tumor-associated or viral antigens. These genetically redirected T-cells display similar anti-tumor or anti-viral activity as the cell clones from which the TCR genes were isolated. However, there is no data describing the activity of T-cells with genetically modified TCR genes specific for intracellular bacterial antigens （e.g., tuberculosis antigens）. This study enriched T-cells specific for the mycobacterial 38 kD glycolipoprotein by repeated in vitro stimulation with antigen-pulsed dendritic cells. Magnetic beads were used to separate CD4^＋ and CD8^＋ T-cells. Spectratype analysis was then used to identify and characterize the third complimentarily-determining regions （CDR3） of the antigen-specific human leukocyte antigen （HLA） class II- and I-restricted TCRs derived from specific CD4^＋ or CD8^＋ T-cells, respectively. Full-length TCR genes were inserted into retroviral vectors and transferred into autologous, primary CD4^＋ and CD8^＋ T-cells. Both redirected CD4^＋ and CD8^＋ T-cells expressed exogenous TCRs with the ability to recognize specific antigens and mediate effector functions. Thus, the method presented in this study generated TCR gene-modified T-cells that can treat multidrug-resistant Mycobacterium tuberculosis infections.