Immune Dysfunction in HIV+ Opioid Users

艾滋病毒感染损害免疫力，即使在持久的病毒学控制之后，免疫扰动仍然存在。这 使用处方类阿片或注射类阿片导致依赖的发生率很高。 已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 对免疫系统来说是灾难性的，但这一领域的研究却严重不足。我们最近的项目 HIV中的免疫力（AI 108472）利用流感疫苗接种作为评估HIV+中免疫力的探针。我们 观察到HIV+比未感染者的疫苗反应更低，并且HIV+药物滥用者亚组 抗体反应比非滥用者差。我们可以将参与者归类为疫苗应答者（VR） 疫苗无应答者（VNR）在研究VNR中免疫缺陷的机制时，我们发现， 外周T滤泡辅助细胞（pTfh）的定量和定性缺陷，pTfh是CD 4 T细胞亚群， 疫苗诱导的抗体反应所必需的细胞。pTfh显示出偏极化 从有利的IL-21分泌表型向有害的IL-2分泌Th 1表型转变， 再加上大量的炎症标志物，导致pTfh不能为B细胞提供所需的免疫原性。 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，并加剧艾滋病毒的免疫功能障碍 即使是在病毒控制的患者中。我们将招募艾滋病毒阴性和病毒感染的阿片类药物使用者和非使用者。 抑制HIV+人群，以解决阿片类药物对抗体反应影响的关键问题 接种季节性流感疫苗后。该项目有三个目标：1。评价炎症的基础状态 以及与pTfh表型和流感疫苗应答的幅度和质量相关的免疫/活化。 2.通过表型、功能和转录组学分析表征pTfh;测试干预措施 用于潜在逆转VNR中阿片类诱导的免疫缺陷。我们将使用单细胞 获得高分辨率数据集的技术，包括多参数流式细胞术和10倍单细胞分析仪。 细胞基因组学平台与使用新的核酸标记的索引表面标志物表达相结合 抗体的为了全面了解阿片类药物对免疫功能的影响， 在存在HIV感染的情况下，我们将使用 无偏基因组学方法来获得与健康对照相比的免疫扰动的快照。 用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价。这些研究是可行的，因为我们 在所描述的技术和访问所需的人口通过我们的ID诊所的专业知识， 针对阿片类药物成瘾人群的专门方案。我们希望提供新的见解， 免疫扰动将有助于制定疫苗方法，包括阿片类疫苗。

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.