ANTIHIV INTERBODIES FOR GENE THERAPY IN CD4+ CELLS

描述：病毒研究所和达纳·法伯癌症研究所 寻求对利用外周设备进行临床前基因转移研究的支持 单个核细胞，从未感染和HIV-1中提纯的CD4+T细胞 将导致“细胞内免疫”的感染者 对抗HIV-1。研究方案将优化大范围 细胞内单链抗体的逆转录病毒基因转移 HIV-1信封，进入临床环境所需的。 载体LNCs105携带选择标记neR和人类 单链抗体F105。载体和包装细胞系已经被 制作。将基因转移到原代细胞的最佳条件是 在HIV-1感染者和非感染者的细胞中建立。 检测HIV-1在长期培养中的复制以评估其能力 抑制HIV-1患者和大鼠细胞中的HIV-1复制 将进行临床规模的CD4+T淋巴细胞转导。基因 细胞的转移、表达、细胞选择和扩增 协议和细胞表面受体的表达水平，形态， 将评估细胞因子谱和对抗原刺激的反应。 DFCI“基因转移设施”和工作人员将参加这次预展 临床ARM和未来的临床建议。

DESCRIPTION: Virus Research Institute and Dana Farber Cancer Institute seeks support for pre-clinical gene transfer studies utilizing peripheral mononuclear cells, purified CD4+ T-cells from uninfected and HIV-1 infected individuals that will result in "Intracellular immunization" against HIV-1. The research proposal will optimize large scale retroviral gene transfer of a intracellular single-chain antibody against the HIV-1 envelope, required for the proceeding into a clinical setting. The vector LNCs105 carries the selection marker neoR and the human single-chain antibody F105. Vectors and packaging cell lines have been produced. Optimal conditions for gene transfer into primary cells will be established in cells from HIV-1 infected and non-infected individuals. Testing HIV-1 replication in long term cultures to evaluate the ability to suppress HIV-1 replication in cells from HIV-1 patients and large clinical scale transduction of CD4+ T lymphocytes will be done. Gene transfer, expression, cell selection and expansion in "dry runs" of the protocol and the level of cell-surface receptor expression, morphology, cytokine profile, and response to antigen stimulation will be evaluated. DFCI "gene transfer facility" and personnel will participate in this pre- clinical arm and in future clinical proposals.