Single cell trancriptomics to identify LTBI reactivation markers in TB/HIV co-infection

项目摘要 结核病的全球控制由于与人类免疫缺陷病毒(HIV)的混合感染而变得更加复杂。 那些感染艾滋病毒的人具有重新激活潜伏性结核病感染(LTBI)的高风险。病毒诱导的慢性免疫 由于SIV合并感染，CD4+T细胞的激活而不是单纯的耗尽与LTBI的重新激活相关。 此外，联合抗逆转录病毒疗法(CART)不能恢复T-效应器功能，从而预防LTBI TB/SIV混合感染的非人灵长类动物模型中的重新激活。CART对慢性阻塞性肺疾病影响的调查 在单细胞水平的相关联合感染临床前模型中的免疫激活将导致识别 预测LTBI重新激活的关键生物标志物。 我们将把这里提出的研究与K01资助的奖项相结合，旨在收集连贯的数据 来自存档的对照样本(未感染、LTBI、SIV感染、结核分枝杆菌/SIV混合感染但幼稚；存档 样本)和Mtb/SIV共同感染的短期、长期CART治疗(K级)恒河猴。基于 我们从联合感染研究中发现，我们假设在结核分枝杆菌/SIV联合感染的病毒血症高峰期启动CART时- 感染导致免疫活性和巨噬细胞周转率明显下降，未能挽救歪斜 导致LTBI重新激活的CD4+T效应器记忆反应。因此，在目标1中，我们将确定具体的 巨噬细胞与CD4+T细胞相互作用中的谱系标记物，尽管手推车进入，但仍受到损害 MTB/SIV混合感染恒河猴。在目标2中，我们将调查SIV驱动的慢性慢性病的最早事件 免疫激活，并将我们的发现与CART治疗的队列进行比较。为此，i)我们将比较激活 血浆细胞样树突状细胞和NK细胞在结核分枝杆菌/SIV混合感染中的地位，II)我们将研究 CART对血浆细胞样树突状细胞(PDC)驱动的I型干扰素(干扰素)产生的影响 与未经处理的对照组相比，下游效应器的反应。 总体而言，我们将能够确定i)巨噬细胞周转是否干扰CD4+T细胞的恢复和 功能，ii)如果CART通过改善NK细胞有效地控制病毒驱动的免疫激活的最早事件 CART在Mtb/SIV共刺激免疫中对I型干扰素应答的影响 感染。研究CART在结核分枝杆菌/SIV混合感染中对免疫激活的影响是识别 用于治疗和疫苗设计的生物标记物，以防止LTBI重新激活。

Project Summary The global control of tuberculosis (TB) is compounded by co-infection with Human Immunodeficiency Virus (HIV). Those infected with HIV are at high risk of reactivating latent TB infection (LTBI). Viral-induced chronic immune activation rather than mere depletion of CD4+ T cells correlates with LTBI reactivation due to SIV co-infection. Further, combinatorial antiretroviral therapy (cART) fails to restore T-effector functions and thus prevent LTBI reactivation in a nonhuman primate model of TB/SIV co-infection. Investigating the impact of cART on chronic immune activation in a relevant co-infected preclinical model at the single cell level will lead to the identification of key biomarkers predicting LTBI reactivation. We will synergize the research proposed here with a funded K01-award aims to collect cohesive data from archived control samples (uninfected, LTBI, SIV infected, Mtb/SIV co-infected but cART naïve; archived samples) and Mtb/SIV co-infected short-term, long-term cART treated (K-award) rhesus macaques. Based on our findings from the co-infection study, we hypothesize that while initiating cART at peak viremia in Mtb/SIV co- infection causes a significant decline in immune activation and macrophage turnover, it fails to rescue the skewed CD4+T effector memory responses leading to LTBI reactivation. Thus, in Aim 1, we will identify the specific lineage markers in the interplay between macrophages and CD4+ T cells that remain impaired despite cART in Mtb/SIV co-infected rhesus macaques. In Aim 2, we will investigate the earliest events of SIV-driven chronic immune activation and compare our findings to cART treated cohorts. For this, i) we will compare the activation status of plasmacytoid dendritic cells (pDCs) and NK cells in Mtb/SIV co-infection, ii) we will study the impact of cART on plasmacytoid dendritic cell (pDC)-driven Type-I Interferon (IFN) production and its impact on downstream effector responses compared to untreated controls. Overall, we will be able to determine if i) macrophage turnover interferes with CD4+ T cell restoration and function, ii) if cART effectively controls the earliest events of virus-driven immune activation by improving NK cell function and iii) the impact of cART on Type-I IFN response in driving the immune activation in Mtb/SIV co- infection. Studying the impact of cART on immune activation in Mtb/SIV co-infection is critical to identifying biomarkers for therapeutics and vaccine design to prevent LTBI reactivation.