ALCOHOL MODULATION OF RECEPTOR SITES FOR HIV AND PRODUCTION OF CHEMOKINES

该提议基于酒精调节的总体假设 肝脏免疫系统在质膜和分子水平上 改变细胞表面受体表达（HIV-1 的结合位点）和 肝非实质细胞中可溶性介质（趋化因子）的产生 细胞[NPC]，即库普弗细胞和窦内皮细胞。 具体来说，急性或慢性酒精调节CD4的表达， 肝NPC上的趋化因子受体和甘露糖特异性受体作为 乙醇诱导肝NPC质膜改变的结果。 该事件还可能导致细胞内 H2O2 的产生增强 进而激活 NF-kappaB。核易位和 转录因子 NF-kappaB 的激活可增强表达 mRNA 的合成以及 α 和 β 趋化因子的合成。这些趋化因子 可能会阻断 HIV-1 的结合或加剧其复制 在CD4淋巴细胞中。 HIV-1 gp120 诱导产生 β-趋化因子 肝性鼻咽癌也可能受到酒精的调节。肝脏是主要的 微生物清除和乙醇代谢的器官。因此，肝细胞 变得容易受到这些可能影响肝脏的物质的影响 本身以及宿主的整体稳态。例如 肝NPC产生的促炎介质增加， 是这些药物的最大贡献者，也可能调节功能 其他器官中的免疫活性细胞。因此，基于这些 综合考虑，提出以下具体目标。具体目标1： 确定急性或慢性酒精中毒对 HIV-1 的影响 肝上 GP120 结合、甘露糖特异性受体和 CD4 的表达 全国人大。肝NPC对HIV-1 gp120的内化和降解 将接受检查。具体目标 2：确定急性或 慢性酒精中毒对细胞内 H2O2 产生的影响， NF-κB 的激活、mRNA 表达和 α（CINC 或 IL-8）和 β（Rantes、MIP-1 α、MIP-1beta、MCP-1）趋化因子 肝NPC。 HIV-1 gp120 对 β-趋化因子的生物活性 将评估产量以及内源性和内源性的作用 外源性内毒素对上述参数的影响（具体目标 1 和 2）。的 后一个目标将检验慢性酒精介导的假设 LPS 从肠道流入循环系统是造成这种情况的原因，至少在 部分，用于调节质膜上的肝 NPC 功能 和分子水平。具体目标 3：确定慢性病的影响 酒精中毒对肝 NPC 功能（即超氧阴离子）的影响 产生、吞噬作用、趋化因子和细胞因子产生、趋化因子 受体（CXCR4 和 CCR5）表达和 SIV gp120 结合。这项研究将 还检查了猿猴模型中趋化因子介导的 SIV-1 复制 SIV-1/艾滋病。酒精对上述过程的总体调节可能 进一步包括HIV-1感染的易感个体和 与这种病毒感染相关的机会病原体。

This proposal is based on the overall hypothesis that alcohol modulates the hepatic immune system at the plasma membrane and molecular levels by altering cell surface receptor expression (binding sites for HIV-1) and production of soluble mediators (chemokines) in hepatic non-parenchymal cells [NPC], i.e., Kupffer cells and sinusoidal endothelial cells. Specifically, acute or chronic alcohol regulates the expression of CD4, chemokine receptors and mannose-specific receptors on hepatic NPC as a result of ethanol-induced alteration in plasma membranes on hepatic NPC. This event could also lead to enhanced intracellular production of H2O2 which in turn activates NF-kappaB. The nuclear translocation and activation of NF-kappaB, a transcription factor, enhances the expression of m-RNA and synthesis of alpha and beta-chemokines. These chemokines could potentially block the binding of HIV-1 or exacerbate its replication in CD4 lymphocytes. The HIV-1 gp120-induced production of beta-chemokines by hepatic NPC may also be modulated by alcohol. The liver is the major organ for microbial clearance and ethanol metabolism. Thus, hepatic cells become susceptible to these agents that could have an impact on the liver itself as well as on the overall homeostasis of the host. For example increased production of pro-inflammatory mediators by hepatic NPC which are the largest contributors of these agents, may also regulate functions of immunocompetent cells in other organs. Thus, based on these considerations, the following specific aims are proposed. Specific aim 1: To determine the effect of acute or chronic alcohol intoxication of hiv-1 GP120 binding, expression of mannose-specific receptors and CD4 on hepatic NPC. The internalization and degradation of HIV-1 gp120 by hepatic NPC will be examined. Specific aim 2: To determine the effect of acute or chronic alcohol intoxication on the intracellular production of H2O2, activation of NF-kappaB, m-RNA expression and secretion of alpha (CINC or IL-8) and beta (Rantes, MIP-1 alpha, MIP-1beta, MCP-1) chemokines in hepatic NPC. The biological activity of HIV-1 gp120 on beta-chemokine production will be assessed, as well as the role of endogenous and exogenous endotoxin on the above parameters (specific aims 1&2). The latter objective will test the hypothesis that chronic alcohol-mediated influx of LPS from the gut to the circulation is responsible, at least in part, for the regulation of hepatic NPC functions at the plasma membrane and molecular levels. Specific aim 3: To determine the effect of chronic alcohol intoxication on hepatic NPC functions, i.e., superoxide anion production, phagocytosis, chemokine and cytokine production, chemokine receptor (CXCR4 & CCR5) expression and SIV gp120 binding. This study will also examine chemokine-mediated SIV-1 replication in a simian model of SIV-1/AIDS. The overall regulation of the above processes by alcohol may further comprise susceptible individuals to HIV-1 infections and opportunistic pathogens associated with this viral infection.