NOVEL SYSTEM TO STUDY MONOCYTE HIV-1 INTERACTION

研究单核细胞 HIV-1 相互作用的新系统

• 批准号: 2443138
• 负责人: Kirk E Sperber
• 金额: $ 12.41万
• 依托单位: ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI
• 依托单位国家: 美国
• 财政年份: 1994
• 资助国家: 美国
• 起止时间: 1994-07-01 至 1999-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2443138
• 关键词: CD3 molecule; MHC class II antigen; antigen presentation; colony stimulating factor; gene expression; genetic techniques; host organism interaction; human genetic material tag; human immunodeficiency virus 1; hybridomas; interferon gamma; interleukin 1; interleukin 8; lipopolysaccharides; macrophage; monocyte; phytohemagglutinins; protein biosynthesis; tissue /cell culture; tumor necrosis factor alpha

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.

我们之前描述了一种研究HIV-1的新系统， 通过感染一系列人单核细胞，单核细胞/巨噬细胞相互作用 和巨噬细胞杂交瘤与各种株的HIV-1。 之一 在这些细胞系中发现的最早的缺陷是它们不能支持 抗原特异性MHC匹配的T细胞增殖反应。 这是 这与1998年的一项研究中描述的最早的免疫缺陷一致， HIV-1感染（缺乏对特定抗原的反应）， 巨噬细胞在疾病过程的早期被感染。 我们的初步 数据表明，抗原特异性应答的丧失实际上可能与 单核细胞功能障碍 在探索这背后的机制时， 由于不能刺激抗原特异性反应， 定义1）II类抗原表达丧失，I类抗原维持 抗原表达，2.）细胞因子产生失调（IL-6增加 和降低的IL-1），和3.）抗原的异常处理。 目标 这一建议的目的是描述这些发现背后的缺陷 并确定细胞内感染是否全面改变分选， 细胞内蛋白质的加工，或者如果病毒有助于 功能下调。 我们将首先确定 HIV-1感染对巨噬细胞中的II类抗原具有抑制作用 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γ-干扰素，GM-CSF， 和LPS处理或用II类的全长cDNA转染 抗原 II类Ag和加工Ag的共定位研究 将使用该模型确定外生的。 慢性的影响 HIV-1感染对抗CD-3和PHA介导的T细胞增殖的影响 也被研究。 我们将评估HIV-1感染对TNF-α的影响。 α、GM-CSF、IL-8和IL-1-α产生。 每种细胞因子水平 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 与II类抗原的研究类似，我们将 确定该缺陷是转录的、转录后的还是 在排序。 这些研究应该有助于我们理解早期的 HIV-1感染中免疫功能紊乱。