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

描述（由申请方提供）：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）的运输到CNS的机制，以及这些细胞的效应功能，这些细胞可能会减轻或加重神经发病机制。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“离体”获得的抗原特异性CD 8 + 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的效应子功能，包括促炎细胞因子（IFN-γ、TNF-α）、溶细胞效应子分子（即，穿孔素、颗粒酶）和MHC I类限制性裂解靶细胞。我们还将评估CSF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名外周血中可检测到HIV特异性CTL的HIV感染患者进行横断面研究;其次，对40名HIV感染患者进行纵向研究，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名临床稳定的未接受HAART的患者和10名具有神经系统疾病症状的患者。我们假设全身淋巴细胞活化（包括LFA-1和VLA-4的上调）导致活化淋巴细胞外渗到CSF中的增加，并且CNS内的病毒复制，随后是局部炎症反应和趋化因子分泌（即，IP-10、MIP-1 α和MIP-1 β、RANTES）以非抗原特异性方式有助于募集表达这些趋化因子受体的细胞。我们还假设鞘内HIV病毒载量和CSF中T细胞募集之间可能存在相关性，并且CSF中CTL的存在可能是神经系统疾病的预测。

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.