Molecular Analysis of HIV-1C Transmission Cofactors

描述（由申请人提供）：尽管目前许多努力都集中在预防HIV-1在非洲的传播，但关于决定传播结果的潜在分子机制的信息仍然很少。虽然已经确定，母婴传播的风险随着母体艾滋病毒血浆RNA的升高而增加，但尚未确定一个病毒阈值，超过该阈值，传播是绝对的，也没有确定一个阈值，低于该阈值，传播不会发生。因此，我们假设HIV-1的传播受到一系列复杂的宿主细胞辅因子的影响，这些辅因子通过影响局部病毒的进入和复制来差异性地促进（或保护）病毒传播。在这个新的调查修订，我们建议评估HIV-1传播的分子相关性，我们称之为“合作者”，以更好地定义围产期传播的决定因素。我们将利用一个优秀的标本库，已建立的传输流体从药物初治，HIV-1C阳性的母亲和他们的婴儿在博茨瓦纳的横断面确定传输辅因子通过RNA表达谱与围产期传播的孕产妇标本。然后，我们将使用实时PCR验证辅因子水平和病毒表达水平。最后，我们将使用HIV-1C和HIV-1B的全基因组分子克隆，在体外评估已鉴定的传播辅因子对HIV-1复制的影响。这些目标旨在确定和功能特性的分子辅助因子对HIV-1的进入和复制的影响，以更好地定义传播的生物决定因素。该提案的一个关键特征是旨在促进波士顿大学医学院（马萨诸塞州波士顿），波士顿大学公共卫生学院和博茨瓦纳-哈佛合作伙伴关系之间的合作。

DESCRIPTION (provided by applicant): Although many efforts are now focused on preventing HIV-1 transmission within Africa, there remains very little information regarding the underlying molecular mechanisms that determine transmission outcome. While it is well established that the risk of mother-to-child-transmission increases with higher maternal HIV plasma RNA, a threshold of virus has not been identified above which transmission is absolute, nor has a threshold been identified below which transmission does not occur. We therefore hypothesize that HIV-1 transmission is influenced by a complex array of host cellular cofactors that differentially promote (or protect from) viral transmission by influencing local viral entry and replication. In this new investigator revision, we propose to evaluate molecular correlates of HIV-1 transmission, which we term, "co, actors", to better define determinants for perinatal transmission. We will utilize an excellent specimen bank that has been established for transmission fluids obtained cross-sectionally from drug naive, HIV-1C positive mothers and their infants in Botswana to identify transmission cofactors through RNA expression profiling of maternal specimens in association with perinatal transmission. We will then validate cofactor levels and viral expression levels using real-time PCR. Finally, we will evaluate the influence of identified transmission cofactors on HIV-1 replication, in vitro, using full genome molecular clones for HIV-1C and HIV-1B. These aims are designed to identify and functionally characterize the influence of molecular cofactors on HIV-1 entry and replication, to better define biological determinants for transmission. A key feature of this proposal is the intent to foster collaboration between the Boston University School of Medicine (Boston, MA), the Boston University School of Public Health and the Botswana-Harvard Partnership This collaborative effort is essential to elucidate the molecular mechanisms involved in HIV-1C transmission and should be considered a research priority.