FUNCTIONAL CONSEQUENCES OF NATURAL HIV-1 ENV GENE DIVERS

发展一个国家的最大困难之一， 有效的艾滋病毒疫苗是逃避保护的潜力 通过产生病毒多样性免疫，特别是在 包膜蛋白 加剧这种担忧的是， 当采集患者血清时， 在感染的早期阶段，对各种 病毒分离物。 虽然主要的中和决定因素， 已经鉴定了包膜表面蛋白基因（“V3环”，大多数 已经被定位到的逃避中和突变中， 日期落在V3之外，表明此 决定因素的位置对它的识别至关重要。 该医师科学家奖旨在为申请人提供 必要的概念背景和实验室培训， 艾滋病毒的生物学，特别强调 分子遗传多样性和生物相关的，功能性的 特色 它的设计是为了允许开发 申请人作为独立的生物医学研究者， 作为一名物理学家兼科学家的研究生涯。 第一阶段将强调分子生物学的学习和研究培训， 病毒学和免疫学。 它将侧重于发展和 设计用于研究抗原性的技术的应用 HIV-1的多样性，包括整个env基因从 几个HIV-1基因组直接从一个人的血液中分离出来， 感染的个体转化到牛痘病毒表达载体中。 的一部分 对病毒基因功能影响的全面研究 第二阶段将涉及env基因的分类， 这些构建体根据它们的中和敏感性， 通过细胞毒性T淋巴细胞识别。 在这些最初的 分类研究，嵌合包膜基因将产生 在抗原性不同的包膜之间绘制中和图 决定簇和T细胞表位。 这些研究也将扩展到其他组织， 个体以及从不同的时间点，当临床 疾病的表现已经改变，以确定决定因素 组织嗜性和毒性。 这种方法也将适用于 一个不同的人，其获得HIV-1的方式不同于 被研究的个体。 这些研究的最终目的是确定 病毒的多样性和个体对病毒的反应，以及 鉴定env内控制关键表位展示的位点， 所有这些都与有效的艾滋病毒疫苗的研制有关。 的 这项建议的教育和培训方面也将使 申请人获得必要的专业知识，以追求在基础研究 包括HIV-1在内的人类逆转录病毒生物学， 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.