CHARACTERIZATION OF CYTOTOXIC RESPONSES TO HIV ANTIGENS

HIV 抗原细胞毒性反应的特征

• 批准号: 3810228
• 负责人: ROBERT T SCHOOLEY
• 金额: --
• 依托单位: WHITEHEAD INSTITUTE FOR BIOMEDICAL RES
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/3810228
• 关键词: AIDS; AIDS vaccines; B lymphocyte; Bacillus Calmette Guerin vaccine; Epstein Barr virus; Escherichia coli; T lymphocyte; affinity chromatography; antibody neutralization test; antiviral antibody; beta galactosidase; cell mediated cytotoxicity; cell transformation; delayed hypersensitivity; genetic manipulation; human immunodeficiency virus; human tissue; leukocyte activation /transformation; mitogens; molecular cloning; monocyte; viral vaccines; virus antigen; virus protein

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.

艾滋病（AIDS）是由 人类免疫缺陷病毒（HIV），一种致病性人类 逆转录病毒已经在世界范围内达到流行病的程度， 需要免疫预防来阻止这种疾病的传播， 至高无上。 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 目前还不清楚什么是保护性免疫，或者 哪些参数决定疾病的临床进展， 已经鉴定了多种HIV特异性免疫应答。 这些包括中和抗体，抗体依赖性细胞免疫， 细胞毒性（ADCC），细胞增殖反应，和 细胞毒性T细胞（CTL）应答。 我们建议系统地 研究免疫原性表位参与产生 这些HIV特异性免疫反应， 重要的免疫原性表位可以被掺入到 亚单位疫苗 重组DNA表达技术将是 怀特黑德研究所的合作者使用它来创建一个 在E.杆菌 大肠杆菌 然后将裂解物或免疫亲和柱纯化的融合蛋白 用作呈递给HIV的克隆抗原的来源- 特异性CTL 三种不同的自体靶细胞方法 将用于抗原呈递。 其中包括a）EBV 永生化的B细胞B）PHA母细胞或c）单核细胞/巨噬细胞。 待使用的效应CTL将从HIV获得。 血清阳性受试者，由a）大量HIV特异性CTL B） HIV预刺激的外周血单核细胞 c）HIV特异性克隆T细胞。 免疫原 然后，表位将用于从血清中除去反应性抗体， 艾滋病毒血清阳性受试者，这些抗体测试他们的 介导ADCC和中和的能力。 的信息 这些研究提供的信息将用于设计一种BCG-HIV 重组疫苗 候选疫苗发现有希望在 然后将在人类临床试验中对动物研究进行测试。