FUNCTIONAL CONSEQUENCES OF NATURAL HIV-1 ENV GENE DIVERS

最大的困难之一被认为是 有效的艾滋病毒疫苗是逃避保护的可能性 通过产生病毒多样性获得免疫力，特别是在 包膜蛋白。加剧这种担忧的是狭隘的、 收集患者血清时病毒类型特定的中和活性 在感染的早期阶段，针对各种不同的 病毒分离物。尽管主要的中和决定因素是 已鉴定出包膜表面蛋白基因(“V3环”，大多数 已经被映射到的逃避中和突变 日期不在V3范围内，这表明此 行列式的放置对其识别是至关重要的。 该医师科学家奖旨在为申请者提供 必要的概念背景和实验室培训 艾滋病毒生物学，特别强调两者之间的相关性 分子遗传多样性与生物相关性和功能性 特点。它的设计目的是允许开发 申请人为独立生物医学调查员，以便 以内科科学家的身份追求研究事业。 第一阶段将强调分子生物学方面的学习和研究培训， 病毒学和免疫学。它将专注于发展和 抗原性研究技术的应用 HIV-1的多样性，包括从中国转移全膜基因 直接从一个人的血液中分离出几个HIV-1基因组 将感染的个体转化为痘苗病毒表达载体。作为以下内容的一部分 病毒基因功能后果的综合研究 多样性，第二阶段将涉及对 这些结构根据它们的中和敏感性和 细胞毒性T淋巴细胞的识别。在这些首字母之后 分类研究，将产生嵌合包膜基因 在抗原发散的包膜之间映射中和 决定因素和T细胞表位。 这些研究也将从相同的组织扩展到其他组织 个体以及来自不同时间点的临床 疾病的表现形式已经改变，以确定决定因素 组织嗜性和致命性。这一方法也将应用于 感染HIV-1的方式不同于 被研究的个人。 这些研究的最终目的是确定抗原性 病毒的多样性和个体内部对病毒的反应，以及 识别环境内控制关键表位显示的位置， 所有这些都与制定有效的艾滋病毒疫苗有关。这个 这项提案的教育和培训方面也将允许 申请人须取得修读基础课程所需的专业知识 人类逆转录病毒的生物学，包括艾滋病毒-1，作为一个独立的 调查员。

One of the greatest difficulties perceived for the development of an effective HIV vaccine is the potential for escape from protective immunity through generation of viral diversity, especially within the envelope protein. Fueling this concern is the demonstration of narrow, virus type-specific neutralizing activity when patient sera, collected during the early stages of infection, are tested against a variety of virus isolates. Although the major neutralizing determinant of the envelope surface protein gene has been identified (the "V3 loop", most of the escape-from-neutralization mutations that have been mapped to date fall outside V3, indicating that the context within which this determinant is placed is critical to its recognition. This Physician Scientist Award is intended to provide the applicant with the necessary conceptual background and laboratory training in the biology of HIV, with specific emphasis on the correlation between molecular genetic diversity and biologically-relevant, functional characteristics. It is designed to allow for the development of the applicant as an independent biomedical investigator in order to enable the pursuit of a research career as a physician-scientist. Phase I will emphasize study and research training in molecular biology, virology, and immunology. It will focus on the development and application of techniques designed to investigate the antigenic diversity of HIV-1 including the transfer of whole env genes from several HIV-1 genomes isolated directly from the blood of a single infected individual into vaccinia virus expression vectors. As part of a comprehensive study of the functional consequences of viral gene diversity, Phase II will involve the classification of the env genes in these constructs according to their neutralization susceptibility and recognition by cytotoxic T lymphocytes. Following these initial classification studies, chimeric envelope genes will be generated between antigenically divergent envelopes to map neutralization determinants and T-cell epitopes. These studies will also be expanded to other tissues from the same individual as well as from different time points when the clinical manifestation of disease has changed in order to identify determinants of tissue tropism and virulence. This approach will also be applied to a different individual whose mode of acquisition of HIV-1 differed from the individual under study. The eventual aim of these studies is to determine the antigenic diversity of virus and response to it within an individual, and to identify sites within env which control display of critical epitopes, all relevant to formulation of an effective HIV vaccine. The educational and training aspects of this proposal will also allow the applicant to gain the expertise necessary to pursue basic studies in the biology of human retroviruses, including HIV-1, as an independent investigator.