Characterizing intact proviral HIV-1 reservoir size and determinants of reservoir dynamics in African populations

项目摘要 抗逆转录病毒治疗无法清除HIV-1感染，因为一个高度稳定的潜伏病毒库 在宿主中持续存在。与HIV-1根除战略有关的关键领域包括潜伏性 水库的建立、规模和组成。目前，对这些关键领域的了解已相当多， B亚型感染的美国男性，但在受影响最严重的人群中， 感染C亚型的南非妇女，或在一般非洲人群中。人口差异可能 存在关于水库的特点，根除战略将需要采取这样的 考虑到差异。因此，在非洲情况下，对HIV-1储存库的描述代表了 一个非常需要关注的领域。 本项目建议首先解决实施高通量、精确储库的需求 通过优化新开发的完整前病毒DNA检测（IPDA）， C亚型HIV-1这一方法将适用于来自夸祖鲁纳塔尔的200多名妇女。水库 这些妇女的体型将与乌干达和美国队列的个体进行比较， 相同的测定来评估群体间的储库差异。本项目还将研究 病毒因子Nef和长末端重复序列，它们是免疫逃避和基因转移的驱动因素。 转录分别在水库的大小和化妆在这些妇女的一个子集。最后，我们将探讨 来自血液和子宫颈的病毒变异体对这些妇女长寿的储存库的贡献， 贝叶斯进化分析。 我们假设Nef介导的MHC-I下调和LTR活性对细胞凋亡有独立的影响。 水库的大小和分布，这些影响根据感染亚型和研究人群而不同。 该项目将允许使用标准化测定方法比较不同人群的储库大小， 将评估建立的大小和动力学的决定因素。

Project Summary Antiretroviral treatment is unable to clear HIV-1 infection because a highly stable latent viral reservoir persists in the host. Key areas of interest with respect to HIV-1 eradication strategies include latent reservoir establishment, size and make-up. A considerable amount is now known about these key areas in subtype B-infected American men, yet there remains limited knowledge in the most affected population, subtype C-infected South African women, or in African populations in general. Population differences may exist with respect to reservoir characteristics, and eradication strategies would need to take such differences into account. Characterization of the HIV-1 reservoir in the African context therefore represents a much needed area of attention. This project proposes to firstly address the need for implementation of a high-throughput, accurate reservoir sizing method in South Africa through optimization of the newly developed intact proviral DNA assay (IPDA) for subtype C HIV-1. This method will be applied to more than 200 women from KwaZulu Natal. Reservoir size in these women will be compared to that of individuals from Ugandan and American cohorts using the same assay to evaulate reservoir differences across populations. This project will also investigate a role for the viral factors Nef and the long terminal repeat, which are drivers of immune evasion and gene transcription respectively, in reservoir size and make-up in a subset of these women. Finally, we will explore the contribution of viral variants from the blood and cervix to the long-lived reservoir in these women using Bayesian evolutionary analyses. We hypothesize that Nef-mediated MHC-I downregulation and LTR activity have independent effects on reservoir size and distribution, and these effects differ according to infecting subtype and study population. This project will allow for comparison of reservoir size across populations using a standardized assay and will evaluate determinants of size and kinetics of establishment.