LIVER AND THE IMMUNODEFICIENCY OF ALCOHOLICS

这项建议的总体目标是通过以下方式阐明这些机制 酒精调节生物活性的几个方面，以及 中性粒细胞和单核巨噬细胞的表面特征，从而 导致了酗酒者的免疫缺陷。具体地说，这 提案将调查造成伤害的机制 通过研究细胞间的相互作用来治疗或消耗乙醇 实质细胞和非实质细胞(库普弗、内皮细胞和 中性粒细胞)。在酒精中毒和内毒素血症期间，这些细胞 类型可能会相互作用，导致免疫反应受损，并 会引起肝脏的组织损伤。这项提案还将调查 中性粒细胞、枯否细胞和内皮细胞功能的调节 与细胞损伤和炎症的发生、发展有关 酒精中毒伴或不伴内毒素血症。它的作用机制 白细胞-内皮细胞-巨噬细胞相互作用与血管内皮细胞释放 相关的细胞因子将在体内和体外进行检测。为了 为了达到这些目标，我将检查趋化因子的释放， 如C5a、白三烯B4、白介素1和8以及肿瘤坏死因子 有没有内毒素血症。巨噬细胞吞噬能力的体外研究 为了与上述趋化因子相互作用，将进行研究。在连接中 有了这些研究，选定配体的受体的调节将是 在酒精中毒和内毒素血症期间进行调查。的后果 体外共培养法检测中性粒细胞或枯否细胞渗出 系统将用来补充前面的研究。此外， 产生超氧阴离子、蛋白质水解酶，如组织蛋白酶G和 弹性蛋白酶将被测定。这些毒剂的破坏性潜力 内皮细胞和肝细胞将在共培养系统中进行研究。 硫代巴比妥酸反应测定脂质 过氧化作用将被用来评估组织或细胞的损害。用于体外实验 将采用细胞毒性试验、~(51)Cr释放试验和ASAT/ALAT试验。 上述实验模型将通过使用 4-甲基吡唑(乙醇脱氢酶抑制剂)乙醛 蛋白水解酶抑制剂、布洛芬(环氧合酶抑制剂)、超氧化物 体内和体外的超氧化物歧化酶和别嘌醇。这些活动的目的是 研究是为了描绘乙醇代谢物，氧气- 衍生自由基、前列腺素和蛋白水解酶与组织损伤和 免疫缺陷。吞噬细胞调节的机制 将使用蛋白激酶C和蛋白激酶C的调节剂来研究其功能 吞噬细胞中的NADPH氧化酶。

The overall objective of this proposal is to elucidate the mechanisms by which alcohol modulates several aspects of the biological activities and surface characteristics of neutrophils and mononuclear phagocytes, thereby contributing to the immunodeficiency of alcoholics. Specifically, this proposal will investigate the mechanisms responsible for injury after ethanol treatment or consumption by studying the cellular interactions between parenchymal and non-parenchymal cells (Kupffer, endothelial cells & neutrophils). During ethanol intoxication and endotoxemia, these cell types may interact to cause impairment of the immune response and to provoke tissue injury in the liver. This proposal will also investigate the modulation of neutrophil, Kupffer and endothelial cell function as they relate to the onset, development of cellular injury and inflammation during ethanol intoxication with or without endotoxemia. The mechanisms of leukocyte-endothelial cell-macrophage interaction and the release of relevant cytokines in vivo and in vitro will be examined. In order to achieve these goals, I will examine the release of chemotactic factors, such as C5a, leukotriene B4, interleukins 1 & 8, and tumor necrosis factor with or without endotoxemia. In vitro studies on the ability of phagocytes to interact with the above chemotaxins will be performed. In connection with these studies the modulation of receptors for selected ligands will be investigated during ethanol intoxication and endotoxemia. Consequences of neutrophil or Kupffer cell extravasation using an in vitro co-culture system will be used to complement the preceding studies. Furthermore the generation of superoxide anion, proteolytic enzymes such as cathepsin G and elastase will be determined. The destructive potential of these agents to endothelial cells and hepatocytes will be studied in a co-culture system. The thiobarbituric acid reaction for the determination of lipid peroxidation will be used to assess tissue or cell damage. For in vitro cytotoxicity assay, 51Cr-release and ASAT/ALAT assays will be employed. The above experimental models will be further manipulated with the use of 4-methypyrazole (inhibitor of alcohol dehydrogenase) acetaldehyde, alpha-1- protease inhibitor, ibuprofen (cyclooxygenase inhibitor), superoxide dismutase and allopurinol in vivo and in vitro. The purpose of these studies is to delineate the contribution of ethanol metabolites, oxygen- derived radicals, prostanoids and proteases on tissue injury and immunodeficiency. The mechanisms underlying the modulation of phagocyte function will be studied with the use of modulators of protein kinase C and NADPH oxidase in phagocytes.