Correlates and Consequences of Increased Immune Activation in HIV + and - IDUs

描述(由申请人提供)：针对RFA-DA-12-009，我们的多学科团队建议研究与注射吸毒(IDU)相关的免疫激活增加的相关性、后果和机制。这项提议的关键是初步数据， 我们是在2009年6月至2011年9月期间进行的一项试点研究中产生的。我们发现，在未感染HIV-1的注射吸毒者中，免疫系统和肠道相关淋巴组织(GalT)的免疫激活水平增加。我们在组织中的发现是新的，尽管已经报道了注射吸毒者的免疫激活，但对相关因素和机制的理解还没有解决。我们的长期目标是使用包括系统生物学在内的多方面方法来探索以下问题：1)与活性IDU相关的血液和组织中免疫活性增强的机制是什么？2)我们能否区分三种可能的因素--无菌注射、阿片类药物的直接影响和慢性丙型肝炎感染--对免疫激活水平的提高的影响？3)当注射停止时，与活性IDU相关的免疫激活增强是可逆的吗？如果是，可逆性的动力学是什么？4)活性注射如何影响联合抗逆转录病毒治疗(CART)中感染HIV-1的静脉吸毒者的免疫重建，我们能否定性和定量地描述差异？为了回答这些复杂的问题，我们制定了一项战略，招募一群HIV-1未感染和感染的活跃注射吸毒者，以及非常仔细匹配的适当对照，我们将从他们那里收集横向和纵向的血液、组织和行为数据。感染HIV-1的活跃注射器将接受CART和 密切监测病毒学和免疫学反应，并与适当的非注射对照组进行比较。我们将分别使用CD38、HLA-DR和Ki67等标记物来测量血液和组织中细胞亚群的激活和增殖水平。还将测量患者血浆中免疫激活的可溶性标记物的水平。包括吸毒行为在内的行为数据将被视为免疫激活和重建的相关因素。为了响应RFA并反映该项目最具创新性的方面，我们将使用系统生物学方法来解决上述问题。我们将利用微阵列对外周血和组织中的特定细胞群进行分类，以进行mRNA转录图谱的分析。此外，我们还将从体外实验中收集细胞进行转录谱分析，以研究选择性阿片类药物的添加和停用对特定T细胞亚群的影响。在洛克菲勒大学临床和翻译科学中心建立的数据分析核心的帮助下，我们将分析转录图谱数据，以识别与免疫激活、免疫重建和注射行为的标志物相关的基因签名和途径。通过研究注射吸毒者与HIV-1和丙型肝炎病毒感染的交集，我们的目标是改善受影响人群和个人的治疗结果。

DESCRIPTION (provided by applicant): In response to RFA-DA-12-009, our multidisciplinary team proposes to study the correlates, consequences, and mechanisms of increased immune activation associated with injection drug use (IDU). Key to this proposal are preliminary data that we generated during a pilot study conducted between June 2009 and September 2011. We discovered increased levels of immune activation systemically and in gut- associated lymphoid tissue (GALT) in HIV-1 uninfected IDUs. Our findings in tissue are novel and though immune activation in IDUs has been reported, an understanding of correlates and mechanism has not been addressed. Our broad term goals are to use a multifaceted approach including systems biology to explore the following questions; 1) What are the mechanisms of increased immune activation in blood and tissue associated with active IDU? 2) Can we separate the effects of 3 possible contributors to increased levels of immune activation- non-sterile injection, direct effects of opiates and chronic infection with Hepatitis C? 3) Is the heightened immune activation associated with active IDU reversible when injection ceases, and if so, what are the dynamics of the reversibility? 4) How does active injection impact immune reconstitution in HIV-1 infected IDUs on combination antiretroviral therapy (cART) and can we characterize differences qualitatively and quantitatively? To answer these complex questions we have devised a strategy to recruit a cohort of HIV- 1-uninfected and -infected active IDUs along with very carefully matched appropriate controls from whom we will collect blood, tissue, and behavioral data both cross-sectionally and longitudinally. HIV-1-infected active injectors will be treated with cART and monitored closely for virologic and immunologic responses and compared to appropriate non-injecting controls. We will measure levels of cellular activation and proliferation using markers such as CD38 and HLA-DR and Ki67 respectively on subsets of cells from blood and tissue. Levels of soluble markers of immune activation will also be measured in patient plasma. Behavioral data including drug use behaviors will be examined as correlates of immune activation and reconstitution. In response to the RFA and reflecting the most innovative aspect of this project, we will use a systems biology approach to address the issues above. We will sort specific cell populations from peripheral blood and tissue for transcriptional profiling of mRNA with the use of microarrays. In addition we will also collect cells from in vitro experiments for transcriptional profiling to study the effects of the addition and withdrawal of select opiates on specific T cell subsets. With the assistance of the Data Analysis Core established by The Rockefeller University Center for Clinical and Translational Science, we will analyze the transcriptional profiling data to identify gene signatures and pathways for correlation with markers of immune activation, immune reconstitution, and injecting behaviors. By studying the intersection of IDU and infection with HIV-1 and HCV, we aim to improve treatment outcomes in affected populations and individuals.