Immune Dysfunction in HIV+ Opioid Users

艾滋病毒感染损害了免疫力，即使在进行了持久的病毒学控制后，免疫紊乱仍然存在。这 人们使用处方类阿片或注射类阿片导致依赖的发生率很高。 众所周知，阿片类药物具有免疫抑制作用，艾滋病毒和阿片类药物的结合可能证明 这对免疫系统来说是灾难性的，但这一领域的研究严重不足。我们最近的项目是关于 艾滋病毒免疫(AI108472)利用流感疫苗接种作为评估艾滋病毒免疫力的探针。我们 观察到艾滋病毒的疫苗反应比未感染的低，艾滋病毒吸毒者的一小部分 抗体反应比非吸毒者差。我们可以将参与者归类为疫苗响应者(VR) 和疫苗无应答者(VNR)。在研究VNR免疫缺陷的机制时，我们发现 外周T滤泡辅助细胞(PTfh)的数量和质量缺陷，这是CD4T的一个亚群 疫苗诱导抗体反应所必需的细胞。PTfh表现出偏振偏振 从有利的IL-21分泌表型转向有害的IL-2分泌Th1表型， 再加上大量的炎症标志物，导致pTfh无法为B细胞提供 分泌抗体所需的辅助信号。我们的中心假设是，长期使用阿片类药物会钝化 免疫反应损害抗体的产生并加剧HIV的免疫功能障碍 即使在病毒控制的患者中也是如此。我们将招募HIV阴性和病毒感染的阿片类药物使用者和非使用者 被抑制的艾滋病毒人群解决阿片类药物对抗体反应影响的关键问题 在接种季节性流感疫苗后。该项目有三个目标：1.评估炎症的基础状态 以及免疫/激活与pTfh表型和流感疫苗反应的大小和质量有关。 2.通过表型、功能和转录分析鉴定pTfh；3.检验干预措施 用于潜在逆转阿片类药物诱导的VNR免疫缺陷。我们将使用单细胞的组合 获得高分辨率数据集的技术，包括多参数流式细胞术和10x Single 结合使用新型核酸标记的指标化表面标记表达的细胞基因组学平台 抗体。为了全面了解阿片类药物对免疫功能的影响这一主题 在存在艾滋病毒感染的情况下，我们将使用以下方法评估感兴趣人群的体外免疫细胞 无偏见的基因组学方法，以获得与健康对照相比的免疫紊乱的快照。 对纯化的细胞亚群和在抗原暴露期间添加吗啡的体外研究将允许 阿片类药物对免疫系统影响的机制评估。这些研究是可行的，因为我们的 在所述技术方面的专业知识，并通过我们的ID诊所和 处理阿片成瘾人群的专门方案。我们希望为我们提供新的见解 免疫紊乱将有助于制定疫苗方法的战略，包括阿片类药物疫苗的方法。

HIV infection impairs immunity and immune perturbations persist even after durable virologic control. This population has a high incidence of using prescription opioids or injection opioids that lead to dependence. Opioids are known to be immunosuppressive, and the combination of HIV and opioids can potentially prove to be disastrous for the immune system, yet this area has been grossly understudied. Our recent project on immunity in HIV (AI108472) utilized influenza vaccination as a probe for assessing immunity in HIV+. We observed that HIV+ had lower vaccine responses than uninfected, and a subgroup of HIV+ drug abusers had worse antibody response than non-abusers. We could classify participants as vaccine responders (VR) and vaccine non-responders (VNR). In studying mechanisms of immune defects in VNR, we identified quantitative and qualitative defects in peripheral T follicular helper cells (pTfh), which are a subset of CD4 T cells that are essential for vaccine-induced antibody responses. The pTfh displayed a skewed polarization away from a favorable IL-21 secreting phenotype towards a detrimental IL-2 secreting Th1 phenotype, coupled with abundance of inflammatory markers, resulting in failure of pTfh to provide B cells with the helper signals required for secreting antibodies. Our central hypothesis is that chronic opioid use blunts the immune response to impair the generation of antibodies and exacerbates the immune dysfunction of HIV even in virally controlled patients. We will recruit opioid users and non- users in HIV negative and virally suppressed HIV+ populations to address key questions on the effect of opioids on the antibody response following seasonal influenza vaccination. The project has 3 aims: 1. To evaluate basal state of inflammation and immune/activation in relation to pTfh phenotype and the magnitude and quality of flu vaccine response. 2. To characterize pTfh by phenotypic, functional and transcriptomic analyses and 3. To test interventions for potential reversal of opioid induced immune deficiency in VNR. We will use a combination of single cell technologies to gain high resolution datasets including multi-parameter flow cytometry and the 10x single cell genomics platform combined with indexed surface marker expression using novel nucleic acid-tagged antibodies. In order to gain a comprehensive understanding on the topic of opioid effect on immune function in presence of HIV infection, we will evaluate immune cells ex vivo from the populations of interest using unbiased genomics approaches to obtain a snapshot of immune perturbation compared to healthy controls. In vitro studies with purified cell subsets and addition of morphine during antigen exposure will allow for mechanistic evaluation of opioid impact on the immune system. These studies are feasible, given our expertise in the technologies described and access to the required population through our ID Clinics and specialized programs dealing with opioid addicted populations. We expect to provide novel insight into immune perturbations that will help in strategizing vaccine approaches, including those for opioid vaccines.