The HIV Latent Reservoir, Suboptimal Immune Response on Antiretroviral Therapy, and Exogenous Cytokine Therapies

项目摘要/摘要 开始抗逆转录病毒治疗(ART)的HIV阳性患者中，有相当比例的人CD4+T细胞较低 尽管多年来对ART进行了病毒学抑制，但Count的CD4计数仍处于异常低的水平。 这些次优免疫反应者(SOIR)的死亡风险是个体的2-3倍。 其CD4计数随着抗逆转录病毒治疗而适当上升，这种较高的死亡风险将持续十年或更长时间。这个 SOIR中免疫恢复不佳和死亡率上升的机制仍然很差 已定义。已经出现了一个明显的关联：SOIR的免疫激活水平明显高于 其他接受过艺术治疗的个人。我们知道，通过T细胞受体刺激HIV感染的CD4 结果HIV特异性细胞毒T淋巴细胞产生和识别HIV RNA和蛋白质。这 提示日常对CD4s的抗原刺激可以产生过度的免疫激活 潜伏艾滋病毒负担非常大的个人。与此相一致的是，两种基因频率之间的相关性 抗逆转录病毒治疗中感染的CD4+T细胞和低的CD4计数已经被报道过多次。然而，这些研究 没有控制对CD4的最低点或对ART的时间，所以尚不清楚SOIR是否有较高的感染负担 CD4+T细胞。我们假设，从HIV潜伏库(LR)诱导增加，无论是因为 较大的LR大小或来自LR的诱导性增加，与次优免疫反应相关。LR大小 和诱导性从来没有被同时评估过，但我们将使用有效的新测试来进行评估， 可以区分完整的和有缺陷的艾滋病毒前病毒。我们将确定LR的大小和诱导性 有助于次优免疫反应，以及旨在增加CD4计数的细胞因子疗法 还可以扩展LR。对于目标1，我们将确定血液和淋巴组织中艾滋病毒受体的大小 与使用新的完整前病毒DNA分析(IPDA)的次优免疫反应呈正相关，a 滴状数字聚合酶链式反应分别对SOIR样品中完整和缺陷前病毒进行定量 以及从ACTG纵向链接随机试验中确定的年龄和谷值匹配的对照 研究和巴尔的摩、旧金山和克利夫兰的三个大队列。对于目标2，我们将确定 使用定量病毒是否更容易从潜伏期诱导SOIR中受感染的CD4+T细胞 对SOIR和配对对照的血液样本进行诱导分析。对于目标3，我们将确定是否 增加CD4计数的细胞因子疗法也通过使用IPDA来测量HIV LR的大小来扩大HIV LR 外源性IL-7、IL-15和IL-2治疗HIV的临床试验样本。通过正规的教学培训 以及来自艾滋病毒储存库、艾滋病毒免疫学、临床研究和生物统计学专家的有组织的指导， PI将在艾滋病毒潜伏技术、免疫技术和知识方面发展独特的技能， 统计和翻译研究。这一培训提供了一条通往独立职业生涯的途径 翻译研究人员，研究病毒因素在治疗后艾滋病毒发病机制中的作用。

Project Summary/Abstract A significant percentage of HIV-positive individuals who start antiretroviral therapy (ART) with a low CD4+ T cell count have CD4 counts that plateau at abnormally low levels despite years of virologic suppression on ART. The risk of death for these suboptimal immune responders (SoIRs) is 2-3 times higher than that of individuals whose CD4 count rises appropriately with ART, and this higher risk of death persists for a decade or more. The mechanisms underlying the suboptimal immune recovery and increased mortality rates in SoIRs remain poorly defined. One clear association has emerged: SoIRs have significantly higher levels of immune activation than other ART-treated individuals. We know that stimulation of HIV-infected CD4s through the T cell receptor results in HIV RNA and protein production and recognition by HIV-specific cytotoxic T lymphocytes. This suggests that the usual daily antigenic stimulation of CD4s could produce excess immune activation in individuals with a very large burden of latent HIV. Concordantly, a correlation between the frequency of infected CD4+ T cells and low CD4 counts on ART has been reported several times. However, these studies did not control for CD4 nadir or time on ART, so it is not clear whether SoIRs have a higher burden of infected CD4+ T cells. We hypothesize that increased induction from the HIV latent reservoir (LR), whether because of a larger LR size or increased inducibility from the LR, is correlated with suboptimal immune response. LR size and inducibility have never been simultaneously evaluated, but we will do so using efficient new assays that can discriminate intact from defective HIV proviruses. We will determine whether LR size and inducibility contribute to suboptimal immune response and whether cytokine therapies designed to increase CD4 counts also expand the LR. For Aim 1, we will determine whether the size of the HIV LR in blood and lymphoid tissue is positively correlated with suboptimal immune response using the new intact proviral DNA assay (IPDA), a droplet digital PCR assay that separately quantifies intact and defective proviruses, on samples from SoIRs and age- and nadir-matched controls identified from within the ACTG Longitudinal Linked Randomized Trials study and three large cohorts in Baltimore, San Francisco, and Cleveland. For Aim 2, we will determine whether infected CD4+ T cells of SoIRs are more readily inducible from latency using a quantitative viral induction assay on blood samples from SoIRs and matched controls. For Aim 3, we will determine whether cytokine therapies that increase CD4 count also expand the HIV LR by using the IPDA to measure LR size in samples from clinical trials of exogenous IL-7, IL-15, and IL-2 in treated HIV. Through formal didactic training and structured mentorship from experts in HIV reservoirs, HIV immunology, clinical research, and biostatistics, the PI will develop a unique skillset in HIV latency techniques, immunological techniques and knowledge, statistics, and translational research. This training provides a pathway to an independent career as a translational investigator researching the contribution of viral factors to the pathogenesis of treated HIV.