HUMORAL AND CELLULAR IMMUNE RESPONSE TO HIV FOR VACCINE DEVELOPMENT

该项目旨在描述对人类的免疫反应。 免疫缺陷病毒(HIV)，并鉴定能够 引发保护性免疫。我们正在研究体液和细胞 对HIV-1和HIV-2的免疫反应；猴免疫缺陷研究 病毒是有计划的。我们首先关注体液免疫反应， 调查HIV中和抗体。以前的研究定位于一个主要的 病毒gpl2O的V3环的中和表位。十字 使用匹配的血清和病毒进行扎伊尔队列的中和研究 目前正在对分离株进行V3环序列分析。数量 在有限的地理区域内的“血清型”将被确定。结果 这些研究将有助于设计启发式的共识表位 广谱中和抗体在未来疫苗中的应用。剥削 体外免疫选择的研究使我们能够进一步鉴定 交替中和表位和病毒包膜区域对 传染性。对单一免疫精选变异病毒的广泛分析， 30%的血清中和亲本的血清抵抗中和 病毒，表明跨膜蛋白中的一个点突变 导致构象变化，改变了中和表位 另一个网站。选择和分析其他变体，使用 HIV-1 IIIB和MN原型毒株的感染性分子克隆 ISY HIV-2分离株的分离将有助于描述更多的病毒 包膜区域在病毒-宿主相互作用中很重要。对该计划的评估 感染或免疫猕猴的HIV-2中和抗体反应 正在进行中。这种猕猴模型也被用来识别病毒表位。 这会引发细胞免疫。细胞增殖试验与铬 针对HIV-2基因的细胞毒性T淋巴细胞释放试验 痘苗病毒构建物中的产品以及合成肽 推定的病毒表位的代表。细胞毒性T细胞的克隆 将促进表位的映射，从而引发保护细胞介导的 免疫反应。

This project aims to delineate immunologic responses to the human immunodeficiency viruses (HIVs), and to identify viral subfragments able to elicit protective immunity. We are investigating humoral and cellular immune responses to HIV-1 and HIV-2; studies with simian immunodeficiency virus are planned. We first focused on the humoral immune response, investigating HIV neutralizing antibody. Previous studies localized a major neutralizing epitope to the V3 loop of the viral gpl2O. Cross neutralization studies of a Zairian cohort using matched sera and viral isolates are being analyzed with regard to V3 loop sequences. The number of "serotypes" within a restricted geographic area will be determined. Results of these studies will assist design of a consensus epitope for elicitation of broadly reactive neutralizing antibody in future vaccines. Exploitation of in vitro immune selection has allowed us to pursue identification of alternate neutralizing epitopes and viral envelope regions important for infectivity. Extensive analysis of a single immune selected variant virus, resistant to neutralization by 30% of sera which neutralize the parental virus, has suggested that a point mutation in the transmembrane protein caused a conformational change, altering a neutralization epitope at another site. Selection and analysis of additional variants using the infectious molecular clones of the IIIB and MN prototype HIV-1 isolates and of the ISY HIV-2 isolate will allow delineation of additional viral envelope regions important in viral-host interactions. Assessment of the HIV-2 neutralizing antibody response in infected or immunized macaques is underway. This macaque model is also being used to identify viral epitopes which elicit cell mediated immunity. Cell proliferation assays and chromium release assays for cytotoxic T-lymphocytes are directed against HIV-2 gene products in vaccinia constructs as well as synthetic peptides representative of putative viral epitopes. Cloning of cytotoxic T-cells will facilitate mapping of epitopes which elicit protective cell mediated immune responses.