CHARACTERIZATION OF CYTOTOXIC RESPONSES TO HIV ANTIGENS

由艾滋病引起的获得性免疫缺陷综合征 人类免疫缺陷病毒(HIV)，一种致病的人类 逆转录病毒在世界范围内已经达到流行的程度， 需要免疫预防来阻止这种疾病的传播 派拉蒙。疫苗的合理设计需要彻底的 了解HIV感染的免疫生物学。虽然它是 目前还不清楚什么是保护性免疫，或者 什么参数决定了疾病的临床进展？ 已经确定了多种HIV特异性免疫反应。 这些包括中和抗体，抗体依赖的细胞 细胞毒性(ADCC)、细胞增殖反应和 细胞毒性T细胞(CTL)反应。我们建议有系统地 研究参与产生的免疫原性表位 这些艾滋病毒特异的免疫反应，以便最 重要的免疫原性表位可以被结合到一个 亚单位疫苗。重组DNA表达技术将成为 由怀特黑德研究所的合作者用来创建一个 大肠杆菌中HIV蛋白重叠克隆文库。大肠埃希菌 裂解产物或免疫亲和柱纯化的融合蛋白 被用作克隆抗原的来源，以呈现给HIV- 特定的CTL。三种不同的自体靶细胞方法 将被用来呈递抗原。其中包括a)EBV 永生化B细胞b)PHA母细胞或c)单核/巨噬细胞。 将使用的效应器CTL将从HIV获得 血清阳性受试者，包括a)大量HIV特异性CTL b) HIV预刺激的外周血单核细胞 体外抗原：c)HIV特异性克隆T细胞。免疫原性 然后将使用表位从血清中洗脱反应性抗体。 艾滋病毒血清阳性的受试者，并对这些抗体进行检测 调解ADCC和中和的能力。这些信息 这些研究提供的信息将被用于设计卡介苗-艾滋病毒 重组疫苗。在中国发现前景看好的候选疫苗 然后，动物研究将在人类的临床试验中进行测试。

The acquired immunodeficiency syndrome (AIDS) induced by the human immunodeficiency virus (HIV), a pathogenic human retrovirus has reached epidemic proportions worldwide, and the need for immunoprophylaxis to stem the spread of this disease is paramount. Rational design of a vaccine requires a thorough understanding of the immunobiology of HIV infection. While it is not yet known exactly what constitutes protective immunity, or what parameters determine clinical progression of disease, a variety of HIV-specific immune responses have been identified. These include neutralizing antibodies, antibody-dependent cellular cytotoxicity (ADCC), cellular proliferative responses, and cytotoxic T cell (CTL) responses. We propose to systemically investigate the immunogenic epitopes involved in generation of these HIV-specific immune responses, in order that the most important immunogenic epitiopes can be incorporated into a subunit vaccine. Recombinant DNA expression technology will be used by collaborators at the Whitehead Institute to create a library of overlapping clones of HIV proteins in E. coli. E coli lysates or immunoaffinity-column purified fusion protein will then be used as a source of cloned antigen for presentation to HIV- specific CTL. Three different autologous target cell approaches will be used for presentation of antigen. These include a) EBV immortalized B cells b) PHA blasts or c) monocyte/macrophages. The effector CTL to be used will be obtained from HIV seropositive subjects and consist of a) bulk HIV-specific CTL b) peripheral blood mononuclear cells prestimulated with HIV antigens in vitro c) HIV-specific cloned T cells. Immunogenic epitopes will then be used to elute reactive antibodies from serum of HIV seropositive subjects, and these antibodies tested for their ability to mediate ADCC and neutralization. The information provided by these studies will then be used in design of a BCG-HIV recombinant vaccine. Candidate vaccines found promising in animal studies will then be tested in clinical trails in humans.