The role of purinergic receptor signaling in the establishment of HIV latency

嘌呤能受体信号传导在 HIV 潜伏期建立中的作用

• 批准号: 279339224
• 负责人: Dr. Ilona Tóth
• 金额: --
• 依托单位: National Institute of Allergy and Infectious Diseases (NIAID)
• 依托单位国家: 德国
• 项目类别: Research Fellowships
• 财政年份: 2015
• 资助国家: 德国
• 起止时间: 2014-12-31 至 2016-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/279339224?language=en
• 关键词: role; purinergic; receptor; signaling; establishment

This application proposes a project to examine the role of purinergic receptor signaling in HIV infection. In particular, the role of ATP and adenosine signaling in the HIV replicative cycle and in the establishment of viral latency will be analyzed. To investigate the role of purinergic receptors (P1 for adenosine, P2X and P2Y for ATP) in HIV entry, replication and latency, in vitro infection assays of healthy, human peripheral blood mononuclear cells with wildtype HIV (JRFL) and reporter HIV virions (R7GEmC, pMM310) are planned in combination with specific purinergic receptor agonists and antagonists. (Provided by Prof. Jacobson, Molecular Recognition Section, NIH). To investigate the role of purinergic receptor signaling in the clinical course of HIV infection,whole transcriptome analysis using next generation sequencing will be carried out on single sorted, HIV-infected CD4+ T cells from HIV-infected people at different stages of disease, both on and off combination anti-retroviral therapy (cART). A droplet-based microfluidic cell sorting procedure for the identification and isolation of HIV-infected cells is already established in the Douek Laboratory. Samples of peripheral blood, lymph node and gut mucosa will be obtained through Dr. Doueks ongoing collaborations with investigators at the University of California San Francisco (Dr. Steven Deeks), Case Western Reserve University (Dr. Michael Lederman), The University of Minnesota (Dr. Timothy Schacker) and through the NIH clinic. Currently, longitudinal samples are available from over 100 HIV-infected individuals. The experimental work will be conducted at the National Institute of Allergy and Infectious Diseases (NIAID) at the campus of the National Institutes of Health (Bethesda, Maryland) in the Human Immunology Section of the Vaccine Research Center (VRC), headed by Daniel Douek M.D., MRCP, Ph.D. Adequate S3 facilities and technical equipment for handling and single-sorting of HIV infected materials and whole transcriptome analysis are accessible in the Genome Analysis Core of the VRC, headed by Daniel Douek.

该申请提出了一个研究嘌呤能受体信号传导在 HIV 感染中的作用的项目。特别是，将分析 ATP 和腺苷信号传导在 HIV 复制周期和病毒潜伏期建立中的作用。为了研究嘌呤能受体（腺苷的 P1，ATP 的 P2X 和 P2Y）在 HIV 进入、复制和潜伏期中的作用，计划与特定的嘌呤能受体激动剂和拮抗剂联合对健康人外周血单核细胞（JRFL）和报告 HIV 病毒体（R7GEmC、pMM310）进行体外感染测定。 （由 NIH 分子识别科 Jacobson 教授提供）。为了研究嘌呤能受体信号传导在 HIV 感染临床过程中的作用，将使用下一代测序对来自不同疾病阶段的 HIV 感染者的单分选的 HIV 感染 CD4+ T 细胞进行全转录组分析，包括联合抗逆转录病毒治疗 (cART) 和非联合抗逆转录病毒治疗 (cART)。 Douek 实验室已经建立了一种基于液滴的微流体细胞分选程序，用于识别和分离 HIV 感染细胞。外周血、淋巴结和肠粘膜样本将通过 Doueks 博士与加州大学旧金山分校（Steven Deeks 博士）、凯斯西储大学（Michael Lederman 博士）、明尼苏达大学（Timothy Schacker 博士）的研究人员持续合作以及通过 NIH 诊所获得。目前，已有超过 100 名 HIV 感染者的纵向样本可供使用。实验工作将在美国国立卫生研究院（马里兰州贝塞斯达）校园内的国家过敏和传染病研究所（NIAID）疫苗研究中心（VRC）人类免疫学部门进行，由 Daniel Douek M.D.、MRCP、Ph.D. 领导。由 Daniel Douek 领导的 VRC 基因组分析核心拥有足够的 S3 设施和技术设备，用于处理和单一分类 HIV 感染材料以及全转录组分析。