Establishment of HIV-specific cytotoxic T cells via TCR gene cloning and gene transfer using in vivo information

HIV特异性细胞毒性T细胞被认为是控制HIV感染的重要因素。然而，在感染HIV的患者中，已经描述了T细胞抗原受体zeta链下调和体外T细胞功能受损。这一现象可能会使艾滋病毒特异性疫苗的开发变得困难。因此，我们试图建立替代性的艾滋病毒特异性免疫疗法。目前，我们还没有足够的策略来操纵抗原特异性T细胞。我们建议将T细胞受体基因转移用于抗原特异性免疫治疗。在提出的技术中，首先要鉴定患者体内重要的抗原特异性T细胞，然后从这些单个T细胞中分离出一对编码α和β T细胞受体的cdna。这些基因随后会被转移到自身淋巴细胞中。这些工程抗原特异性细胞也被操纵表达适当的功能基因，然后可以应用于特异性免疫治疗。为了证明该系统可以在HIV感染中起作用，我们使用了一个实验系统，在BALB/c小鼠中引发了针对HIV env gp160肽P18IIIB的免疫反应。利用TCR克隆分析和基因转移重构TCR功能的信息，证明了通过TCR基因克隆可以获得HIV特异性CTL。

HIV-specific cytotoxic T cells have been regarded to be an important factor to control HIV infection. However, in the patients infected with HIV, T cell antigen receptor zeta chain down-regulation and impaired in vitro T cell function have been described. This phenomenon could potentially make the development of HIV-specific vaccination difficult. We, therefore, tried to establish alternative HIV-specific immunotherapy.At present, we do not have a sufficient range of strategies for manipulating antigen-specific T cells. We propose that T cell receptor gene transfer could be used for antigen-specific immunotherapy. In the proposed technique, important antigen-specific T cells in patients would first be identified, and then a pair of cDNAs encoding alpha and beta T cell receptors would be isolated from these single T cells. These genes would then be transferred into self lymphocytes. These engineered antigen-specific cells also manipulated to express appropriate functional genes could then be applied to specific immunotherapy.In order to prove this system can work in HIV infection, we used an experimental system, in which immune responses against HIV env gp160 pepetide P18IIIB was elicited in BALB/c mice. We demonstrated that HIV specific CTL could be obtained using TCR gene cloning using the information of TCR clonal analysis and reconstitution of the TCR function by gene transfer.