Opioid Modulation of Immunocompetence

描述：（由申请人提供）本竞争性续期建议书是 旨在确定小鼠免疫系统中哪些细胞表达6，g 和K阿片受体，以及这些细胞的激活是否会改变 阿片受体的表达和多种阿片受体的分布 在不同的淋巴细胞群体中。通过使用荧光阿片类药物， 流式细胞仪，我们已经开发了一种新的间接荧光方法， 检测关于：阿片受体。这种方法比 放射受体结合方法学。双标记实验表明， 小鼠胸腺细胞和巨噬细胞上的最高水平的about受体表达， 和人类小胶质细胞。本申请中提出的研究重点是 对β和β阿片受体的检测。根据功能研究， 第一个要检验的假设是小鼠胸腺细胞和CD 4+和CD 8 + T细胞 细胞表达;5阿片受体。为了检测6种受体，两种方法将 被比较。6-选择性配体纳曲吲哚的FITC标记衍生物， 将以类似于用 FITC结合的K配体。第二种方法将使用特异性抗体 针对6受体。第二个要检验的假设是， 小鼠腹膜和T细胞表达β-阿片样物质受体。g受体 将用FITC标记的芬太尼或吗啡衍生物标记， 和/或针对Ia阿片受体的抗体。流式细胞术将 用于检测标记的受体。对于6和g受体， 是开发一种检测受体的最佳方法，然后使用 这种方法来研究受体的表达。第三个假设是， 淋巴细胞的活化将增加阿片受体的表达 因为已经显示受体mRNA的水平随着细胞活化而增加。 总的来说，这些研究将使用创新的方法和技术， 确定哪些免疫细胞表达大约：，6，和大约阿片样物质 受体以及受体表达是否随着细胞活化而改变。 这些研究将为确定细胞机制奠定基础 阿片类药物通过其改变免疫功能和对病毒的免疫应答 感染，包括艾滋病。

DESCRIPTION: (provided by applicant) This competitive renewal proposal is directed at determining which cells from the mouse immune system express 6, g and K opioid receptors, and whether activation of these cells alters the amount of receptor expressed and the distribution of the multiple opioid receptors among the different lymphocyte populations. By using fluorescent opioids and flow cytometry, we have developed a novel indirect fluorescent method for detecting about: opioid receptors. This method is more sensitive than radioreceptor binding methodology. Double-labeling experiments have shown the highest level of about receptor expression on mouse thymocytes and macrophages, and on human microglial cells. The studies proposed in this application focus on the detection of 6 and/.t opioid receptors. Based on functional studies, the first hypothesis to be tested is that mouse thymocytes and CD4+ and CD8+ T cells express ;5 opioid receptors. To detect 6 receptors, two approaches will be compared. FITC-labeled derivatives of the 6-selective ligand, naltrindole, will be used in a manner similar to the labeling of the receptor with the FITC-conjugated K ligand. The second approach will use specific antibodies directed against the 6 receptor. The second hypothesis to be tested is that mouse peritoneal and T cells express the/,t opioid receptor. The g receptor will be labeled with either a FITC-labeled derivative of fentanyl or morphine, and/or antibodies directed against the la opioid receptor. Flow cytometry will be used to detect the labeled receptors. For both 6 and g receptors, the goal is to develop an optimal method for detecting the receptor, and then to use this method to study receptor expression. The third hypothesis is that activation of lymphocytes will increase the expression of opioid receptors since levels of receptor mRNA have been shown to increase with cell activation. Collectively, these studies will use an innovative approach and technique to determine which cells from the immune express about:, 6, and about opioid receptors and if receptor expression is altered with activation of the cells. These studies will lay the foundation for determining the cellular mechanisms by which opioids alter immune function and immunological responses to viral infection, including AIDS.