OPIATE RECEPTOR-MEDIATED EFFECTS OF STRESS ON IMMUNITY

阿片剂和阿片肽可以改变免疫功能。因此，接触到 鸦片类药物可能是决定艾滋病毒感染易感性的辅助因素。这 应用重点是细胞和分子研究，旨在阐明 阿片肽影响免疫的基本机制 功能。正在调查的机制的两个要素是：(1) 纳洛酮耐药的理化和功能表征 正常免疫细胞上可能介导许多作用的结合部位 (2)在正常免疫细胞内诱导编码的mRNAs 对于前脑啡肽-A和前阿片黑素皮质素， 脑啡肽和B-内啡肽。纳洛酮耐药特性的研究 B-内啡肽结合部位将涉及放射性配基结合研究 确定Kd、Bmax、竞赛特征、条件 表达正常人结合位点的诱导和细胞亚群 脾细胞和胸腺细胞制剂。物理化学表征 将涉及[125I]-B-内啡肽与结合的共价交联 网站，然后进行分析的十二烷基硫酸钠-PAGE。蔗糖密度梯度分析 在没有交联剂的情况下，也将执行。建立职能部门 依赖刀豆蛋白A的胸腺细胞这个结合部位的意义 将采用增殖试验和胸腺细胞辅助细胞试验 特征：(1)结合部位的诱导；(2)剂量依赖性 结合部位的激活；(3)通过以下方式选择性地激活结合部位 B-内啡肽和其他阿片类药物的片段；和(4)细胞亚群 它们是结合位点激活的最接近的靶标。阿片类药物的研究进展 胸腺细胞和脾细胞中肽mRNA的表达将集中在 小鼠肝炎病毒(MHV)A59、Aa感染的诱导作用 天然的鼠类病原体。剂量-反应、动力学关系、细胞 受影响的亚群，活与灭活MHV的需求将是 在活体内测定。体外实验将评估 IL-1b介导MHV对mRNA表达的影响。最后， 免疫组织化学研究将评估阿片类药物翻译产品 在MHV感染后存在于免疫细胞中。这些研究都是关于 纳洛酮耐药受体与阿片类药物编码基因的表达 多肽将有助于定义旁分泌系统，免疫细胞由此产生 对阿片肽有反应。

Opiates and opioid peptides modify immune function. Thus, exposure to opiates may be a cofactor determining susceptibility to HIV infection. This application focuses on cellular and molecular studies designed to elucidate basic mechanisms underlying the influence of opioid peptides on immune function. Two elements of the mechanism under investigation are: (1) the physicochemical and functional characterization of naloxone-resistant binding sites on normal immune cells which may mediate many of the effects of B-endorphin; (2) induction within normal immune cells of mRNAs coding for prepro-enkephalin-A and proopiomelanocortin, precursors to the enkephalins and B-endorphin. Characterization of naloxone-resistant B-endorphin binding sites will involve radioligand binding studies to determine the Kd, Bmax, competition characteristics, conditions of induction and cell subpopulations expressing the binding site from normal splenocyte and thymocyte preparations. Physicochemical characterization will involve covalent crosslinking of [125 I]-B-endorphin to the binding site followed by analysis on SDS-PAGE. Sucrose density gradient analysis without crosslinking will also be performed. To establish the functional significance of this binding site, a concanavalin-A dependent thymocyte proliferation assay and thymocyte helper cell assay will be used to characterize: (1) induction of the binding site; (2) dose-dependence of binding site activation; (3) selective activation of the binding site by fragments of B-endorphin and other opioids; and (4) cell subpopulations which are proximate targets of binding site activation. Studies of opioid peptide mRNA expression in thymocytes and splenocytes will focus on the inductive effects of infection with murine hepatitis virus (MHV) A59, a natural murine pathogen. The dose-response, kinetic relations, cell subpopulations affected, and requirement for live versus killed MHV will be determined in vivo. In vitro experiments will evaluate whether interleukin-lB mediates the effect of MHV on mRNA expression. Finally, immunohistochemical studies will assess whether opioid translation products are present in immune cells after MHV infection. These studies of the naloxone-resistant receptor and the expression of mRNAs coding for opioid peptides will help define a paracrine system whereby immune cells produce and respond to opioid peptides.