OPIOID MODULATION OF IMMUNOCOMPETENCE

描述：(申请人摘要) 这项竞争性的更新建议旨在确定哪些小区 来自小鼠和人类的免疫系统表达Mu、Delta和kappa阿片类物质 感受器。进一步的研究旨在确定药物滥用、艾滋病毒 感染和艾滋病改变阿片受体在脑内的分布 人类淋巴细胞。我们已经开发了一种间接荧光方法来检测 检测阿片类药物结合部位。这种方法比其他方法更灵敏 放射受体结合方法学。双标记实验使用 针对特定CD细胞表面的荧光标记抗体 多种阿片受体的标记物及其标记 荧光素偶联阿片类药物，随后进行放大程序将 结果确定了哪种类型的白细胞表达哪种类型的(S) 阿片受体。初步数据显示，kappa受体是 在R1EGO胸腺瘤细胞系、小鼠胸腺细胞和 脾细胞和CD4+胸腺细胞。类似的实验将会进行。 用荧光素标记Mu和Delta类阿片。建议数 实验将导致本地化的彻底表征 小鼠Mu、Delta和kappa受体的相对数量 淋巴细胞。药理学证据表明，T细胞的激活 增加阿片受体的表达。小鼠胸腺细胞、巨噬细胞 而纯化的CD4+和CD8+细胞将被激活和受体数量 而分布将被比作静止的细胞。这些实验将 探讨淋巴细胞的激活是否会改变 阿片受体及其分布，有助于确定其功能 免疫细胞上的多个阿片受体。研究将集中于 关于静脉注射药物滥用、艾滋病毒感染和艾滋病是否会改变 阿片受体的表达或分布。 拟议的实验将最终确定哪些细胞来自 免疫系统表达经典的阿片受体，如果滥用药物，艾滋病毒 感染和艾滋病改变了这些受体的表达。

DESCRIPTION: (Applicant's Abstract) This competitive renewal proposal is directed at determining which cells from the mouse and human immune systems express mu, delta, and kappa opioid receptors. Studies are further directed at determining if drug abuse, HIV infection and AIDS alters the number of distribution of opioid receptors on human lymphocytes. We have developed an indirect fluorescent method for detecting opioid binding sites. This method is more sensitive than radioreceptor binding methodology. Double labeling experiments using fluorescently tagged antibodies directed against specific CD cell surface markers and labeling the multiple opioid receptors with fluorescein-conjugated opioids, followed by an amplification procedure will result in determining which types of leukocytes express which type(s) of opioid receptors. Preliminary data demonstrate that the kappa receptor is expressed on the R1EGO thymoma cell line, on mouse thymocytes and splenocytes, and on CD4+ thymocytes. Similar experiments will be performed with mu and delta opioids labeled with fluorescein. The proposed experiments will result in a thorough characterization of the localization and relative number of the mu, delta and kappa receptors on mouse lymphocytes. Pharmacological evidence suggests that activation of T cells increases the expression of opioid receptors. Mouse thymocytes, macrophages and purified CD4+ and CD8+ cells will be activated and the receptor number and distribution will be compared to resting cells. These experiments will address whether activation of lymphocytes alters the expression of the opioid receptor and its distribution, and will help determine the function of the multiple opioid receptors on immune cells. Studies will be focused on determining if intravenous drug abuse, HIV infection and AIDS alters the expression or distribution of either mu, delta, or kappa opioid receptors. The proposed experiments will definitively determine which cells from the immune system express the classical opioid receptors and if drug abuse, HIV infection and AIDS alters the expression of these receptors.