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的一个 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得主要资金， 因此可以在其他CRISP条目中表示。列出的机构是 中心，不一定是研究者的机构。 为了研究HIV达特特异性胞内抗体对病毒感染的保护作用，成年恒河猴（Macaca mulatta）作为抗HIV-1感染和AIDS发展的基因治疗实验的模型。 在再输注表达不相关胞内抗体（A3 H5-Flag）和治疗性抗tat sFv胞内抗体（hutat 2-HA）的离体MLV转导的富含CD 4+（CD 8耗尽）的细胞后，用致病性SHIV毒株静脉内攻击动物。通过FACS和实时PCR分别分析转导的CD 4+细胞的存活和A3 H5或sFv hutat 2转基因的基因标记。 还监测了处理的猕猴中SHIV血浆RNA水平和CD 4计数。此外，还将监测针对A3 H5或hutat 2胞内抗体的CTL和/或抗体。对腋窝淋巴结进行连续采样，并对LN结构进行组织病理学评价。最后，还将通过IF鉴定转导的细胞的细胞内A3 H5或sFvhutat 2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.