Model System Studies of Naltrexone and Alcohol Interaction

描述(申请人提供)：纳曲酮用于治疗包括酒精中毒在内的药物成瘾。纳曲酮在控制酒精的免疫抑制作用方面的潜在用途最近已被确定。特别是，纳曲酮增强了阿片配体对自然杀伤细胞功能的影响。这些观察到的潜在分子基础尚不清楚。我们认为，通过Mu和Delta阿片受体之间的物理联系来调节受体功能是一种潜在的机制。我们还假设纳曲酮对u阿片受体的占位足以增加配体结合和诱导自然杀伤细胞中的增量阿片受体的信号的能力。为了验证这些假说，我们建议通过免疫沉淀技术鉴定自然杀伤细胞膜上同时表达阿片受体或表达突变的mU受体的阿片受体免疫复合体，以检验自然杀伤细胞中u阿片受体和Delta阿片受体的物理联系。我们建议通过使用不同的分子和生化技术来评估阿片受体抑制对自然杀伤细胞中阿片受体结合水平、阿片受体信号转导以及细胞毒因子和细胞因子产生的影响，从而确定阿片受体寡聚的生理后果。我们计划通过检测乙醇诱导的自然杀伤细胞阿片受体免疫复合体水平、配体结合、细胞毒因子和细胞因子的产生和信号的变化，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的改变。还将通过检测阿片受体的物理联系和药理学变化以及NK细胞的阿片配体反应来评估纳曲酮预防乙醇引起的阿片受体功能偶联改变的能力。这项研究产生的信息应该有助于我们开发一种创新的策略，通过异二聚体结合阿片类激动剂和拮抗剂来治疗酒精中毒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.