HIV-1 Clones with Attenuated Cytopathic Effects

描述（申请人提供）：HIV在感染个体中的发病机制和组织培养中的细胞病变效应与病毒载量和复制密切相关。我们建议研究几个独特的HIV-1克隆，这些克隆具有高病毒载量的长期非进展性细胞病变效应（CPE），这将使我们能够从HIV-1发病机制的决定因素中分离出病毒载量和复制的决定因素。大多数来自长期非进展者（LTNP）的HIV-1分离株是CCR5-tropic (R5)，并且在体内和组织培养中复制较差，相反，一些LTNP具有中等到高的病毒载量（LTNP- hvl），与进展为艾滋病的个体相当。我们假设并有初步数据表明，一些LTNP-HVL衍生的HIV-1克隆与进展为艾滋病的个体衍生的HIV-1克隆一样具有复制能力，但细胞病变程度较低。本应用程序中提出的研究将使用这些独特的克隆来定义所有HIV-1克隆的遗传决定因素和发病机制。这些独特的HIV-1克隆可能使我们能够将病毒发病机制的决定因素与病毒复制的决定因素分离开来，这可能会影响艾滋病治疗方法和疫苗的开发。我们有以下具体目标：表征LTNP-HVL R5 HIV-1分离株在FTOC、扁桃体组织培养和SCID-hu小鼠中的复制和细胞病变效应。我们将进一步表征两个我们已经开始研究的具有减毒CPE的HIV-1克隆，并将在FTOC和扁桃体组织培养中表征至少两个这些独特且信息丰富的HIV-1生物克隆。最弱的HIV-1克隆也将在SCID-hu小鼠中进行研究。2. 鉴定LTNP-HVL中R5 HIV-1分离株的Env和Nef相关表型。这些表型将被研究，因为它们与HIV-1的发病机制有关，因为它们在HIV-1克隆从进展和非进展以及从感染的早期到晚期之间存在差异。

DESCRIPTION (provided by applicant): HIV pathogenesis in infected individuals and cytopathic effects in tissue culture are tightly correlated with viral load and replication. We propose to study several unique HIV-1 clones with attenuated cytopathic effects (CPE) derived from long term non progressors with high viral load, which will allow us to separate the determinants of viral load and replication from the determinants of HIV-1 pathogenesis. Most HIV-1 isolates from long-term non-progressors (LTNP) are CCR5-tropic (R5) and replicate poorly in vivo and in tissue culture, in contrast, some LTNP paradoxically have moderate to high viral load (LTNP-HVL), equivalent to that seen in individuals who progress to AIDS. We hypothesize and have preliminary data which indicate, that some LTNP-HVL derived HIV-1 clones are equally replication competent but less cytopathic than HIV-1 clones from individuals who progress to AIDS. The studies proposed in this application will use these unique clones to define the genetic determinants and mechanisms of pathogenesis of all HIV-1 clones. These unique HIV-1 clones may allow us to separate determinants of viral pathogenesis from determinants of viral replication, which may impact the development of AIDS therapeutics and vaccines. We have the following specific aims: 1. Characterize the replication and cytopathic effects, of R5 HIV-1 isolates from LTNP-HVL in FTOC, tonsil histoculture and SCID-hu mice. We will further characterize two HIV-1 clones with attenuated CPE that we have started to study and will characterize at least two more of these unique and informative HIV-1 biological clones in FTOC and tonsil histoculture. The most attenuated HIV-1 clones will also be studied in SCID-hu mice. 2. Characterize the Env and Nef associated phenotypes of R5 HIV-1 isolates from LTNP-HVL in tissue culture. These phenotypes will be studied since they have been linked to pathogenesis of HIV-1 due to their variation between HIV-1 clones from progressors and non-progressors and from early to late stages of infection.