CHARACTERIZATION OF CYTOTOXIC RESPONSES TO HIV ANTIGENS

HIV 抗原细胞毒性反应的特征

• 批准号: 3803672
• 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/3803672
• 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 line; cell mediated cytotoxicity; cell transformation; delayed hypersensitivity; genetic manipulation; human immunodeficiency virus; human tissue; leukocyte activation /transformation; mitogens; molecular cloning; monocyte; neutralizing antibody; recombinant DNA; 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.

由艾滋病引起的获得性免疫缺陷综合症（AIDS） 人类免疫缺陷病毒（HIV），一种人类致病性病毒 逆转录病毒已在全球范围内达到流行程度， 需要免疫预防来阻止这种疾病的传播 最重要的。 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 尚不确切知道什么构成保护性免疫力，或者 哪些参数决定疾病的临床进展， 已经确定了多种艾滋病毒特异性免疫反应。 这些包括中和抗体、抗体依赖性细胞 细胞毒性（ADCC）、细胞增殖反应和 细胞毒性 T 细胞 (CTL) 反应。 我们建议系统地 研究参与产生的免疫原性表位 这些艾滋病毒特异性免疫反应，以便最 重要的免疫原性表位可以整合到 亚单位疫苗。 重组DNA表达技术将 怀特海研究所的合作者使用它来创建 大肠杆菌中 HIV 蛋白重叠克隆文库。 大肠杆菌 然后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 特定的CTL。 三种不同的自体靶细胞方法 将用于抗原的呈递。 其中包括 a) EBV 永生化 B 细胞 b) PHA 母细胞或 c) 单核细胞/巨噬细胞。 所使用的效应CTL将从HIV获得 血清反应呈阳性的受试者由 a) 大量 HIV 特异性 CTL b) 组成 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 然后使用表位从血清中洗脱反应性抗体 HIV 血清阳性受试者，并对这些抗体进行了测试 介导 ADCC 和中和的能力。 信息 这些研究提供的结果将用于设计 BCG-HIV 重组疫苗。 候选疫苗被发现有希望 然后动物研究将在人类临床试验中进行测试。