CELLULAR ASPECTS OF OPIATE BINDING TO LEUKOCYTES

外源性吗啡类药物和内源性阿片类药物的作用 内啡肽，在人类细胞免疫系统的直接调节中 在传染病方面有潜在的重要后果。都是非法的 药物成瘾和应激导致的β-内啡肽释放已被引用 作为增加对传染病易感性的因素， 普通感冒到肺结核和艾滋病。而疫苗的免疫学效应 免疫细胞暴露在鸦片类药物中是众所周知的，几乎没有信息 关于吗啡对免疫的作用机制已有研究 细胞。在中枢神经系统中，一类G蛋白偶联阿片剂 受体的存在解释了各种鸦片类药物的活性。 最近从这两只小鼠身上克隆了这些受体的基因 和人类来源及其结构-7-跨膜螺旋的结构 片段蛋白推定。然而，当免疫细胞通过 逆转录酶/聚合酶链式反应方法，只有Delta 并发现了kappa受体。Mu受体(MOR)，初级兴奋 根据配体判断，中枢神经系统中的吗啡亲和结合部位缺失 结合实验和RT-PCR检测MOR基因的表达。事实上，唯一的 植物血凝素上存在高亲和力的吗啡结合位点 活化的T淋巴细胞。(-)-吗啡可以通过以下方式从该地点取代 (-)-和(+)-吗啡和B-内啡肽的N-端，但不是通过 Dago，DADLE和U-50488。因为这是唯一报道的，高亲和力 T淋巴细胞上吗啡的位置，重要的是克隆这个位置并 确定它是否仅存在于激活的细胞类型中，或者是否 也存在于低于检测水平的未激活细胞上 结合试验的阈值。除了诱导性之外，它还有可能 在诱导之前，这个位点也可能被合乎宪法地表达 可作为吗啡对T细胞作用的第一步 淋巴细胞。为了验证这一假设，POR将通过与 已知的阿片受体使用RT-PCR或通过表达克隆。POR 然后，克隆将用于研究通过以下方式分离的淋巴细胞群 基于其激活状态的磁珠/抗体络合。 淋巴细胞的激活状态也将被特定的 有丝分裂原刺激。这些实验的结果将回答2 免疫药理学的重要问题--1.诱导吗啡 结合部位(POR)足以解释吗啡的作用 细胞免疫系统？；和2.表达之间有什么相关性 T细胞的诱导结合部位和激活状态 亚型？最终，为了控制鸦片类药物对免疫系统的影响 系统，有必要了解阿片类药物的机制 调节细胞免疫系统。目前，在以下方面存在重大差距： 我们对免疫系统中阿片类药物机制的了解 拟议中的实验将会结束。

The role of exogenous morphine-based drugs and endogenous opioids, like B- endorphin, in the direct modulation of the human cellular immune system has potentially important consequences in infectious disease. Both illicit drug addiction and stress-induced release of B-endorphin have been cited as factors in elevated susceptibility to communicable diseases from the common cold to TB and AIDS. While the immunological effects of the exposure of immune cells to opiates are well known, little information has been obtained concerning the mechanism of action of morphine on immune cells. In the central nervous system, a family of G protein-coupled opiate receptors exist which account for the activity of a variety of opiates. The genes for these receptors have recently been cloned from both murine and human sources and their structure - that of a 7-transmembrane helical segment protein - deduced. However, when immune cells are screened by reverse transcriptase/polymerase chain reaction methods, only the delta and kappa receptors were found. The mu receptor (MOR), the primary high affinity morphine binding site in the CNS, is missing as judged by ligand binding assays and RT-PCR assays for expressed MOR mRNA. In fact, the only high affinity morphine binding site (POR) is found on phytohemagglutin activated T lymphocytes. (-)-Morphine can be displaced from this site by both (-)- and (+)-morphine and the N-terminal of B-endorphin, but not by DAGO, DADLE and U-50,488. Because this is the only reported, high affinity site for morphine on T lymphocytes, it is important to clone this site and determine whether it is present only on activated cell types or whether it is also present on nonactivated cells at levels below the detection threshold of the binding assays. Beside being inducible, it is possible that this site might also be expressed constituitively prior to induction and could function as the initial step in morphine's action on T lymphocytes. To test this hypothesis, POR will be cloned by homology with known opiate receptors using RT-PCR or by expression cloning. The POR clone will then be used to investigate lymphocyte populations isolated by magnetic bead/antibody complexation based on their state of activation. The activation state of the lymphocytes will also be modified by specific mitogen stimulation. The results of these experiments will answer 2 important question of immunopharmacology - 1. Is the inducible morphine binding site (POR) sufficient to explain the action of morphine on the cellular immune system?; and 2. What is the correlation between expression of the induced binding site and the activation state of the T cell subtype?. Ultimately, to control the effects of opiates on the immune system, it is necessary to understand the mechanisms by which opiates modulate the cellular immune system. Currently, there is a major gap in our knowledge about opiate mechanisms in the immune system which the proposed experiments would close.