Mechanisms Underlying the HIV-HSV-2 Syndemic

HIV-HSV-2 综合征的潜在机制

• 批准号: 10063474
• 负责人: Betsy C. Herold
• 金额: $ 48.81万
• 依托单位: ALBERT EINSTEIN COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-12-05 至 2022-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10063474
• 关键词: AIDS prevention; Affect; African; Binding Sites; Biological; Biopsy; CCR5 gene; CD4 Positive T Lymphocytes; CRISPR/Cas technology; CXCR4 gene; Caring; Cell physiology; Cells; Chronic; Contralateral; Development; Disease Outbreaks; Disease Progression; Enrollment; Epidemic; Epidemiology; Flow Cytometry; Frequencies; Gene Expression; Genetic Transcription; Genital; Genitalia; Goals; HIV; HIV Infections; HIV Long Terminal Repeat; Helper-Inducer T-Lymphocyte; Human Herpesvirus 2; Immune; Immunity; In Situ; In Vitro; Individual; Infection; Inflammation; Inflammatory; Inflammatory Response; Knock-out; Lesion; Libraries; Link; Longitudinal Studies; Maintenance; Microbicide Trials Network; Molecular; Participant; Pathway interactions; Peripheral Blood Mononuclear Cell; Pharmacology; Phenotype; Phytohemagglutinins; Plasma; Play; RUNX1 gene; Recombinants; Risk; Role; Sampling; Shock; Side; Signal Pathway; Simplexvirus; Site; Skin; Small Interfering RNA; Stimulus; T-Lymphocyte; T-Lymphocyte Subsets; Testing; Viral; Viral Load result; Virus; Woman; Women' s Interagency HIV Study; antiretroviral therapy; biobank; co-infection; cofactor; cohort; cytokine; epidemiology study; immune activation; immune function; men; novel; peripheral blood; pre-exposure prophylaxis; prevent; programmed cell death protein 1; repository; response; seropositive; small molecule; transcriptome sequencing; transmission process; trial comparing

The HIV and HSV-2 syndemic is well recognized, but the biological mechanisms that contribute are not understood. The recent recognition that HSV-2 is characterized by a frequent state of subclinical shedding suggests that the virus might contribute to persistent immune activation and prompted us to examine the impact of HSV-2 on peripheral blood T cells and on HIV reservoirs. Taking advantage of our biorepository of peripheral blood mononuclear cells (PBMC) from well-characterized HIV+ women on antiretroviral therapy who were or were not HSV-2 seropositive (HIV+/HSV-2+ vs. HIV+/HSV-2-), we found a significant difference in the phenotype of CD4+ (but not CD8+) T cells in HIV+/HSV-2+ compared to HIV+/HSV-2- women. These changes included an increase in the frequency of activated cells, but a paradoxical decrease in the expression of IL-32, an intracellular cytokine presumed to be associated with inflammation. Moreover, when CD4+ T cells isolated from virally suppressed HIV+/HSV-2+ women were stimulated with latency reversal agents, the addition of recombinant IL-32 to the cultures blocked HIV reactivation. These observations suggest that IL-32 plays a pivotal role in controlling HIV reactivation and suggest a new paradigm underlying the HIV-HSV-2 syndemic. We hypothesize that HSV-2 triggers changes in local (at the site of HSV-2 genital skin outbreaks) and peripheral blood CD4+ T cells including a reduction in intracellular IL-32 levels that promote HIV reactivation. Conversely, high levels of IL-32 contribute to the maintenance of HIV reservoirs suggesting that IL-32 blockade may synergize with strategies to reactivate HIV as part of a “shock and kill” approach to cure. To test these hypotheses, we will analyze serial samples of PBMC from HIV infected women before and after HSV-2 acquisition from two unique cohorts: women enrolled in Microbicides Trial Network (MTN)-015, a longitudinal study of African women who seroconverted to HIV while participating in pre-exposure prophylaxis trials, and U.S. women with established HIV infection enrolled in the Bronx Women's Interagency Study (WIHS). We will also compare PBMC in HIV-infected men who are or are not coninfected with HSV-2. We will phenotype immune cell subpopulations to define the changes that occur in association with HSV-2 acquisition and the impact of these changes on plasma viral loads and HIV reservoirs. We will prepare CD4+ T cell libraries of IL- 32lo cells and determine whether these subpopulations are enriched in HSV-2 and/or HIV reactive cells and whether decreased IL-32 interferes with immune functions. We will also take advantage of our repository of genital skin biopsies (herpes lesion and unaffected contralateral side) and analyze the CD4+ T cells to determine whether they are enriched for cells of specific phenotypes in situ. We will determine how IL-32 blocks the response to latency reactivating stimuli and how IL-32 antagonists promote HIV reactivation. These studies will identify pathways and molecules that could be targeted to block HIV reactivation in response to HSV-2 or conversely, enhance latency reversal as part of “shock and kill” HIV eradication strategies.

HIV 和 HSV-2 综合征已广为人知，但其生物学机制尚不明确。 最近认识到 HSV-2 的特点是频繁的亚临床脱落状态。 表明该病毒可能有助于持续的免疫激活，并提示我们检查 利用我们的生物储存库研究 HSV-2 对外周血 T 细胞和 HIV 病毒库的影响。 外周血单核细胞 (PBMC) 来自接受抗逆转录病毒治疗的特征明确的 HIV + 女性 无论是否为 HSV-2 血清阳性（HIV+/HSV-2+ 与 HIV+/HSV-2-），我们发现了显着差异 与 HIV+/HSV-2- 女性相比，HIV+/HSV-2+ 女性的 CD4+（而非 CD8+）T 细胞表型发生这些变化。 包括激活细胞频率的增加，但 IL-32 表达的反常减少， 此外，当分离出 CD4+ T 细胞时，推测与炎症相关的细胞内细胞因子。 来自病毒抑制的 HIV+/HSV-2+ 女性接受潜伏逆转剂刺激，添加 重组IL-32 加入到培养物中可阻止HIV 再激活。这些观察结果表明IL-32 发挥着重要作用。 在控制 HIV 重新激活方面发挥着关键作用，并提出了 HIV-HSV-2 综合征的新范例。 我们探索了 HSV-2 引发局部（HSV-2 生殖器皮肤爆发部位）和 外周血 CD4+ T 细胞包括促进 HIV 再激活的细胞内 IL-32 水平降低。 离线时，高水平的 IL-32 有助于维持 HIV 储存库，表明 IL-32 阻断 可能与重新激活艾滋病毒的策略协同作用，作为“休克和杀死”治疗方法的一部分来测试这些方法。 假设，我们将分析 HSV-2 前后 HIV 感染女性的 PBMC 连续样本 从两个独特的群体中获得：参加微生物杀灭剂试验网络 (MTN)-015 的女性，这是一项纵向研究 对参加暴露前预防试验时血清转化为艾滋病毒的非洲妇女的研究，以及 已确诊感染艾滋病毒的美国妇女参加了布朗克斯妇女机构间研究 (WIHS)。 我们还比较了感染或未感染 HSV-2 的 HIV 感染男性的 PBMC。 免疫细胞亚群来定义与 HSV-2 获取相关的变化以及 这些变化对血浆病毒载量和 HIV 病毒库的影响 我们将准备 IL- 的 CD4+ T 细胞库。 32lo 细胞并确定这些亚群是否富含 HSV-2 和/或 HIV 反应性细胞以及 IL-32 是否会干扰免疫功能 我们还将利用我们的资源库。 生殖器皮肤活检（疱疹病变和未受影响的对侧）并分析 CD4+ T 细胞以 确定它们是否原位富集特定表型的细胞 我们将确定 IL-32 如何进行。 阻断对潜伏期重新激活刺激的反应以及 IL-32 拮抗剂如何促进 HIV 重新激活。 研究将确定可靶向阻止艾滋病毒重新激活的途径和分子 HSV-2 或相反，增强潜伏期逆转，作为“休克和杀死”HIV 根除策略的一部分。