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，因为高度稳定的潜伏病毒库 在宿主中持续存在。在根除艾滋病毒-1战略方面感兴趣的主要领域包括潜伏 水库的建立、大小和组成。现在人们对这些关键领域有相当多的了解 感染B亚型的美国男性，但在受影响最严重的人群中，知识仍然有限， 感染C亚型的南非妇女，或整个非洲人口。人口差异可能 在储层特征方面存在，根除战略将需要采取这样的 考虑到差异。因此，在非洲背景下对艾滋病毒-1宿主的描述代表了 这是一个非常需要关注的领域。 该项目建议首先解决实施高吞吐量、准确水库的需求 南非最新建立的完整前病毒DNA测定法(IPDA)的优化 对于C亚型HIV-1。这一方法将应用于夸祖鲁纳塔尔的200多名女性。水库 这些女性的体型将与乌干达和美国队列的个体进行比较，使用 用相同的方法来评估不同种群之间的储集层差异。该项目还将调查 病毒因子Nef和长末端重复序列是免疫逃避和基因的驱动因素 在这些女性中的一小部分中，转录分别是在储存库大小和构成上。最后，我们将探索 来自血液和宫颈的病毒变异对使用这些药物的妇女的长寿储存库的贡献 贝叶斯进化分析。 我们假设Nef介导的MHC-I下调和LTR活性在 这些影响因感染亚型和研究人群的不同而不同。 该项目将允许使用标准化的化验方法比较不同人群的水库大小 将评估建立规模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.