ANTI-HIV-1 TAT HUMAN SFV INTRABODY GENE THERAPY AGAINST SHIV IN RHESUS MACAQUES

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 为了研究HIV TAT特异性细胞内抗体对病毒感染的保护作用，以成年恒河猴(Macaca Mulatta)为模型，进行了抗HIV-1感染和艾滋病发生发展的基因治疗实验。在重新输注体内表达无关抗体(A3H5-Flag)和治疗性抗TAT SFV抗体(hutal2-HA)的体外MLV转导的CD4(CD8缺失)细胞后，动物被致病的SHV毒株静脉攻击。分别用流式细胞仪和实时荧光定量聚合酶链式反应分析转导的CD4细胞的存活率和A3H5或SFV hutal2转基因基因的基因标记。还监测了SIV病毒血浆RNA水平和治疗后猕猴的CD4计数。此外，还将监测针对A3H5或HUTA2体内的CTL和/或抗体。腋窝淋巴结将被连续采样，并评估LN结构的组织病理学。最后，还将通过IF鉴定转导细胞是否有细胞内A3H5或sFvhutal2的表达。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. To study protection effects of an HIV Tat specific intracellular antibody against viral infection, adult rhesus monkeys (Macaca mulatta) served as a model for gene therapy experiments against HIV-1-infecton and the development of AIDS. Following re-infusion of ex-vivo MLV transduced CD4+-enriched (CD8-depleted) cells, expressing an irrelevant intrabody (A3H5-Flag) and a therapeutic anti-tat sFv intrabody (hutat2-HA), animals were challenged intravenously with a pathogenic SHIV strain. Survival of transduced CD4+ cells and gene marking of the A3H5 or sFv hutat2 transgenes were analyzed, respectively, by FACS and real-time PCR. Levels of SHIV plasma RNA and CD4 counts in the treated macaques were also monitored. In addition, CTLs and/or antibodies against the A3H5 or hutat2 intrabodies will also be monitored. Axillary lymph nodes will be sampled serially and evaluated histopathologic for LN architecture. Finally, transduced cells will also be identified by IF for intracellular A3H5 or sFvhutat2 expression.