NOVEL SYSTEM TO STUDY MONOCYTE HIV-1 INTERACTION

我们之前已经描述了一种研究HIV-1的新系统- 感染一系列人单核细胞的单核细胞/巨噬细胞相互作用 和各种HIV-1毒株的巨噬细胞杂交瘤。其中一个 这些细胞系最早的缺陷是它们不能支持 抗原特异性MHC与T细胞增殖反应相匹配。这是 中描述的最早的免疫缺陷之一 艾滋病毒-1感染(对特定抗原缺乏反应)和 巨噬细胞在病程早期被感染。我们的预赛 数据表明，抗原特异性反应的丧失实际上可能与 单核细胞功能障碍。在探索这一现象背后的机制 不能刺激抗原特异性反应，有三个缺陷 已定义1。)在维持I类的情况下II类抗原表达丢失 抗原表达，2)细胞因子产生失调(IL-6增加 并降低IL-1)和3。抗原的异常处理。目标是 这项建议的目的是描述这些发现背后的缺陷 并确定全球范围内的细胞内感染是否改变了分类和 细胞内蛋白质的处理或病毒是否参与 功能下调。我们将首先确定达到的级别 HIV-1感染对巨噬细胞中II类抗原的抑制作用 杂交瘤细胞系(转录或在蛋白质生产中分选)。 然后我们将尝试用伽马干扰素，GM-CSF， 和脂多糖处理或全长c DNA转染人II类病毒 抗原。第II类银与已加工银的共定位研究 外生性将用这个模型来确定。慢性精神分裂症的影响 HIV-1感染对抗CD-3和PHA介导的T细胞增殖的影响 也有待研究。我们将评估HIV-1感染对肿瘤坏死因子的影响。 产生α、GM-CSF、IL-8和IL-1-α。各细胞因子水平 将在慢性感染的细胞系中与未感染的细胞系进行比较 细胞系。类似于对II类抗原的研究，我们将 确定缺陷是转录缺陷、转录后缺陷还是 在分类方面。这些研究应该有助于我们理解早期的 HIV-1感染中的免疫功能紊乱。

We have previously described a novel system to study HIV-1- monocyte/macrophage interactions by infecting a series of human monocyte and macrophage hybridomas with various strains of HIV-1. One of the earliest defects noted in these cell lines was their inability to support an antigen specific MHC matched T cell proliferative response. This is consistent with one of the earliest immunologic deficits described in HIV-1 infection (lack of response to specific antigens) and the fact that macrophages are infected early in the course of disease. Our preliminary data suggested that loss of antigen specific responses may in fact relate to monocyte dysfunction. In exploring the mechanisms underlying this inability to stimulate an antigen specific response, three defects were defined 1.) loss of Class II antigen expression with maintained Class I antigen expression, 2.) dysregulated cytokine production (increased IL-6 and decreased IL-1), and 3.) abnormal processing of antigen. The goal of this proposal is to characterize the defects underlying these findings and determine whether intracellular infection globally alters sorting and processing of intracellular proteins or if virus contributes to downregulation of function. We will first determine the level at which HIV-1 infection is inhibitory for Class II antigens in the macrophages hybridoma cell lines (transcription or in protein production of sorting). We will then attempt to reverse the defect with gamma-interferon, GM-CSF, and LPS treatment or transfection with full length cDNA for the Class II antigens. Co-localization studies of Class II Ag and Ag processed exogenously will be determined using this model. The effect of chronic HIV-1 infection on anti-CD-3 and PHA mediated T cell proliferation will also be studied. We will assess the effect of HIV-1 infection on TNF- alpha, GM-CSF, IL-8 and IL-1-alpha production. Levels of each cytokine will be compared in the chronically infected cell lines to the uninfected cell lines. Similar to the studies for the Class II antigens, we will determine whether the defect is transcriptional, post-transcriptional or in sorting. These studies should aid in our understanding of early immune dysfunctional in HIV-1 infection.