OPIATE RECEPTOR-MEDIATED EFFECTS OF STRESS ON IMMUNITY

阿片类药物和阿片肽改变免疫功能。因此，暴露于 阿片类药物可能是决定HIV感染易感性的辅助因素。这 应用程序侧重于细胞和分子研究，旨在阐明 阿片肽影响免疫功能的基本机制 功能所调查的机制的两个要素是： 纳洛酮耐药细胞理化和功能特性 正常免疫细胞上的结合位点，其可介导许多效应 （2）在正常免疫细胞内诱导编码β-内啡肽的mRNA 前脑啡肽原-A和阿黑皮素原， 脑啡肽和β-内啡肽。纳洛酮耐药的表征 β-内啡肽结合位点将涉及放射性配体结合研究， 确定Kd、Bmax、竞争特性、 诱导和细胞亚群表达的结合位点从正常 脾细胞和胸腺细胞制备物。物理化学表征 将涉及[125 I]-B-内啡肽与结合的共价交联 位点，然后在SDS-PAGE上分析。蔗糖密度梯度分析 也将进行无交联的反应。建立功能性 这个结合位点的意义，伴刀豆球蛋白-A依赖的胸腺细胞 增殖试验和胸腺细胞辅助细胞试验将用于 表征：（1）结合位点的诱导;（2） 结合位点活化;（3）通过选择性活化结合位点， β-内啡肽和其他阿片类物质的片段;以及（4）细胞亚群 其是结合位点活化的邻近目标。阿片类药物研究 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，a感染的诱导作用 自然鼠病原体。剂量反应，动力学关系，细胞 受影响的亚群，以及对活MHV与灭活MHV的要求， 在体内测定。体外实验将评估是否 白细胞介素-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翻译产物是否 在MHV感染后存在于免疫细胞中。这些研究 纳洛酮耐药受体与阿片样物质基因表达 肽将有助于定义免疫细胞产生的旁分泌系统， 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.