Model System Studies of Naltrexone and Alcohol Interaction

描述（由申请人提供）：纳洛酮用于治疗药物成瘾，包括酒精中毒。最近已经确定了纳洛酮在控制酒精的免疫抑制作用中的潜在用途。特别地，纳洛酮增强阿片样物质配体对自然杀伤细胞功能的作用。这些观察结果的潜在分子基础尚不清楚。我们提出，μ和δ阿片受体之间的物理关联的受体功能的调制是一个潜在的机制。我们还假设纳洛酮对μ阿片受体的占用足以增加配体结合和诱导自然杀伤细胞中δ阿片受体信号传导的能力。为了检验这些假设，我们建议检查物理协会的μ阿片受体和δ阿片受体在自然杀伤细胞中，通过识别的免疫复合物的阿片受体的自然杀伤细胞的膜表达阿片受体或表达突变的μ受体，使用免疫沉淀技术。我们建议通过评估μ阿片受体对δ阿片受体结合水平的抑制作用，δ阿片受体的信号传导，以及使用各种分子和生物化学技术在自然杀伤细胞中产生细胞毒性因子和细胞因子，来确定阿片受体寡聚化的生理后果。我们计划通过确定乙醇诱导的阿片受体免疫复合物、配体结合、信号传导和自然杀伤细胞中细胞毒性因子和细胞因子产生水平的变化来评估乙醇抑制NK细胞对内源性阿片配体的反应是否涉及阿片受体寡聚化的改变。纳洛酮预防乙醇诱导的阿片受体功能偶联改变的能力也将通过测量阿片受体的物理缔合和药理学变化以及NK细胞的阿片配体反应来评价。从这项研究中产生的信息应该帮助我们开发一种创新的策略，通过异二聚体结合阿片激动剂和拮抗剂来治疗酒精和其他免疫缺陷患者的免疫功能不全。

DESCRIPTION (provided by applicant): Naltrexone is used for the treatment of drug addiction including alcoholism. A potential use of naltrexone in controlling the immune-suppressive effect of alcohol has recently been identified. In particular, naltrexone enhances the effects of opioid ligands on natural killer cell functions. The underlying molecular basis for these observations is not known. We propose that the modulation of receptor function by physical association between mu and delta opioid receptors is a potential mechanism. We also hypothesize that the occupancy of mu opioid receptors by naltrexone is sufficient to increase the ability of a ligand to bind and induce signaling of delta opioid receptors in natural killer cells. To test these hypotheses, we propose to examine the physical association of mu opioid receptors and delta opioid receptors in natural killer cells by identifying the immune complex of opioid receptors in the membrane of the natural killer cells expressing both opioid receptors or expressing mutated mu receptors using immunoprecipitation techniques. We propose to determine the physiological consequences of opioid receptor oligomerization by evaluating the effects of repression of mu opioid receptors on the levels of delta opioid receptor binding, the signaling by delta-opioid receptors, and the production of cytotoxic factors and cytokines in natural killer cells using various molecular and biochemical techniques. We plan to evaluate whether ethanol inhibition of the NK cell response to endogenous opioid ligands involves alteration of opioid receptor oligomerization by determining the ethanol-induced changes in the levels of opioid receptors immunocomplex, ligand binding, signaling and production of cytotoxic factors and cytokines in natural killer cells. The ability of naltrexone to prevent ethanol-induced alteration of opioid receptor functional coupling will also be evaluated by measuring the changes in the physical association and pharmacology of opioid receptors and the opioid ligands responses of NK cells. The information generated from this study should help us to develop an innovative strategy for combining opioid agonists and antagonists by heterodimeric association to treat immune incompetence in alcoholic and other immune-deficient patients.