Extracellular vesicles in semen and genital HIV infection and immunity during heroin addiction and methadone or buprenorphine substitution therapy

 描述（由申请人提供）：我们最近已经表明，人类精液含有数万亿个细胞外囊泡（SEV），其携带可以调节免疫系统的小细胞外RNA（exRNA）分子（Nucleic Acids Research，42：7290-7304，2014）。在未发表的研究中，我们发现SEV抑制适应性T细胞反应，最有可能是通过影响抗原呈递。SEV还与体外的HIV病毒粒子相关联，并且在我们手中增强了HIV易感性，尽管另一个小组最近报道了相反的情况。最终，SEV可能会通过直接与HIV及其靶细胞相互作用， 削弱了T细胞对艾滋病病毒的保护性免疫。阿片类药物也会影响免疫系统，尽管确切的机制和功能结果还不清楚。海洛因滥用导致免疫功能低下，增加了艾滋病毒的易感性，但行为和生物学原因很难解开。海洛因成瘾通常用美沙酮或丁丙诺啡长期药物治疗，这两种阿片类药物的免疫学特性和对艾滋病毒感染的影响也尚不清楚。因此，精液和阿片类药物使用中细胞外囊泡的相互作用可能对HIV感染和免疫系统产生重要影响，本提案将研究这些联系。我们将确定SEV对阴道外植体模型中HIV感染的影响，并剖析SEV介导的适应性T细胞应答（包括HIV疫苗接种）抑制的性质和程度。我们将在正常供体的血液和精液样本中进行HIV易感性，适应性T细胞反应和SEV生物学（exRNA组成和功能）的比较研究，以及开始辅助美沙酮或丁丙诺啡治疗前和三个月后的海洛因使用者。这三种阿片类药物也将在培养物中进行测试，因为阴道白细胞上阿片类药物受体样1受体的表达（我们的数据）以及其他非阿片类药物受体如Toll样受体4的报告表明阿片类药物可直接影响白细胞生物学。SEV和阿片样物质的相互作用将在组合实验中解决。这些研究将加深我们对精液中阿片类药物和细胞外囊泡单独或共同影响性传播艾滋病毒的理解。了解这些影响将有助于改善艾滋病毒疫苗接种的结果，关注其他艾滋病毒预防策略，并提供美沙酮与丁丙诺啡替代疗法的临床益处的新信息。

 DESCRIPTION (provided by applicant): We have recently shown that human semen contains trillions of extracellular vesicles (SEV) carrying small extracellular RNA (exRNA) molecules that may regulate the immune system (Nucleic Acids Research, 42:7290-7304, 2014). In unpublished studies, we have found that SEV suppress adaptive T cell responses, most likely by affecting antigen presentation. SEV also associate with HIV virions in vitro and in our hands enhance HIV susceptibility, although another group recently reported the opposite. Ultimately, SEV likely impact HIV infection, both by directly interacting with HIV and its target cells, and by compromising protective T cell immunity against HIV. Opioids too affect the immune system, although the exact mechanisms and functional outcomes are not well- understood. Heroin abuse leads to an immunocompromised state and increases HIV susceptibility, but the behavioral and biological causes for this are difficult to untangle. Heroin addiction is often treated with long- term methadone or buprenorphine medication, two opioids whose immunological properties and effects on HIV infection also remain unclear. Thus, the interplay of extracellular vesicles in semen and opioid use may have important consequences for HIV infection and the immune system, and this proposal will investigate these connections. We will determine the effects of SEV on HIV infection in a vaginal explant model and dissect the nature and extent of SEV- mediated inhibition of adaptive T cell responses, including to HIV vaccination. We will perform comparative studies of HIV susceptibility, adaptive T cell responses and SEV biology (exRNA composition and function) in blood and semen samples from normal donors, as well as heroin users before and three months after starting assisted methadone or buprenorphine treatment. The three opioids will also be tested in culture, because expression of the Opioid Receptor-Like 1 receptor on vaginal leukocytes (our data), and reports of other non- opioid receptors such as Toll-like receptor 4, indicate that opioids could directly affect leukocyt biology. SEV and opioid interactions will be addressed in combinatorial experiments. These studies will deepen our understanding of how opioids and extracellular vesicles in semen, alone or in conjunction, impact sexual HIV transmission. Knowledge of these effects will help to improve HIV vaccination outcomes, focus other HIV prevention strategies, and provide new information on the clinical benefits of methadone versus buprenorphine substitution therapy.