HIV-Specific T-cells in cerebrospinal fluid (CSF)

脑脊液 (CSF) 中的 HIV 特异性 T 细胞

• 批准号: 6605853
• 负责人: Barbara L. Shacklett
• 金额: $ 2.81万
• 依托单位: J. DAVID GLADSTONE INSTITUTES
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-07-03 至 2003-10-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6605853
• 关键词: AIDS /HIV neuropathy; AIDS therapy; HIV infections; cell migration; central nervous system; cerebrospinal fluid; chemokine; clinical research; cytokine receptors; cytotoxic T lymphocyte; cytotoxicity; enzyme linked immunosorbent assay; flow cytometry; human subject; interferon gamma; leukocyte activation /transformation; longitudinal human study; neuroimmunomodulation; patient oriented research; plasma; tumor necrosis factor alpha; virus replication

DESCRIPTION (provided by applicant): This proposal responds to PA-01-072, "HIV-1 Infection of the Central Nervous System (CNS)." This study has two major goals. The first goal is to elucidate the mechanisms driving the trafficking of immune effector cells, particularly cytotoxic T-cells (CTL), to the CNS, and the effector functions of these cells that may either palliate or exacerbate neuropathogenesis. The second goal is to determine the relationship between trafficking of antiviral T-cells to cerebrospinal fluid (CSF) and viral load, antiretroviral therapy, and development of neurological disease. In preliminary studies, we determined that multiparameter flow cytometry may be used to detect antigen-specific CD8+ T-cells obtained "ex vivo" from 5-10 ml of CSF. In Specific Aim 1, we will study the frequency and trafficking patterns of HIV-specific CTL in CSF, and determine the relationship of CTL frequency in CSF to systemic and intrathecal immune activation, chemokine expression, HIV viral load and development of CNS disease. In Specific Aim 2, we will assess the effector functions of CSF CTL, including secretion of proinflammatory cytokines (IFN-gamma,, TNF-alpha), cytolytic effector molecules (i.e., perforin, granzymes), and MHC class I restricted lysis of target cells. We will also assess the clonality of HIV-specific CTL in CSF and peripheral blood, and determine the extent of"overlap" between these two populations. Two studies will be performed: first, a cross-sectional study of 25 HIV-infected patients with detectable HIV-specific CTL in perpheral blood; second, a longitudinal study of 40 HIV-infected patients including 10 who are stopping or interrupting highly-active antiretroviral therapy (HAART), 10 who are beginning HAART, 10 clinically stable individuals not on HAART and 10 with symptoms of neurological disease. We hypothesize that systemic lymphocyte activation (including upregulation of LFA-1 and VLA-4) leads to increased extravasation of activated lymphocytes into CSF, and that viral replication within the CNS, followed by a local inflammatory response and secretion of chemokines (i.e., IP-10, MIP-1alpha and MIP-1beta,RANTES) contributes to recruitment of cells expressing receptors for these chemokines in a non antigen-specific manner. We also hypothesize that there may be a correlation between intrathecal HIV viral load and T-cell recruitment to CSF, and that the presence of CTL in CSF may be predictive of neurological disease.

描述（由申请人提供）：本提案响应 PA-01-072“中枢神经系统 (CNS) 的 HIV-1 感染”。这项研究有两个主要目标。第一个目标是阐明驱动免疫效应细胞（特别是细胞毒性 T 细胞 (CTL)）向 CNS 运输的机制，以及这些细胞可能减轻或加剧神经发病的效应功能。第二个目标是确定抗病毒 T 细胞向脑脊液 (CSF) 的运输与病毒载量、抗逆转录病毒治疗和神经系统疾病的发展之间的关系。在初步研究中，我们确定多参数流式细胞术可用于检测从 5-10 ml CSF 中“离体”获得的抗原特异性 CD8+ T 细胞。在具体目标1中，我们将研究CSF中HIV特异性CTL的频率和运输模式，并确定CSF中CTL频率与全身和鞘内免疫激活、趋化因子表达、HIV病毒载量和CNS疾病发展的关系。在具体目标 2 中，我们将评估 CSF CTL 的效应功能，包括促炎细胞因子（IFN-γ、TNF-α）的分泌、溶细胞效应分子（即穿孔素、颗粒酶）和 MHC I 类限制性靶细胞裂解。我们还将评估脑脊液和外周血中HIV特异性CTL的克隆性，并确定这两个群体之间的“重叠”程度。将进行两项研究：首先，对 25 名 HIV 感染者进行横断面研究，这些患者的外周血中可检测到 HIV 特异性 CTL；其次，一项针对 40 名 HIV 感染者的纵向研究，其中包括 10 名正在停止或中断高效抗逆转录病毒治疗 (HAART) 的患者、10 名正在开始 HAART 的患者、10 名未接受 HAART 且临床稳定的患者以及 10 名有神经系统疾病症状的患者。我们假设全身淋巴细胞激活（包括 LFA-1 和 VLA-4 的上调）导致激活的淋巴细胞外渗至 CSF 的增加，并且中枢神经系统内的病毒复制以及随后的局部炎症反应和趋化因子（即 IP-10、MIP-1α 和 MIP-1beta、RANTES）的分泌有助于招募 以非抗原特异性方式表达这些趋化因子受体的细胞。我们还假设鞘内 HIV 病毒载量与 CSF 中 T 细胞募集之间可能存在相关性，并且 CSF 中 CTL 的存在可能预示着神经系统疾病。