Model System Studies of Naltrexone and Alcohol Interaction

说明（由申请人提供）：纳曲酮用于治疗药物成瘾，包括酒精中毒。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。这些观察的潜在分子基础尚不清楚。我们认为通过阿片受体和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。我们还假设纳曲酮占据阿片受体足以增加配体结合和诱导自然杀伤细胞中阿片受体信号传导的能力。为了验证这些假设，我们建议通过使用免疫沉淀技术鉴定表达阿片受体或表达突变的mu受体的自然杀伤细胞膜中的阿片受体的免疫复合物，来检查mu阿片受体和δ阿片受体在自然杀伤细胞中的物理关联。我们建议使用各种分子和生化技术，通过评估抑制mu阿片受体对δ阿片受体结合水平、δ阿片受体信号传导以及自然杀伤细胞中细胞毒性因子和细胞因子产生的影响，来确定阿片受体寡聚化的生理后果。我们计划通过测定乙醇诱导的自然杀伤细胞中阿片受体免疫复合物、配体结合、信号传导和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细胞对内源性阿片受体配体的反应是否涉及阿片受体寡聚化的改变。纳曲酮防止乙醇诱导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关联和药理学变化以及NK细胞的阿片配体反应来评估。从这项研究中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.