CELLULAR ASPECTS OF OPIATE BINDING TO LEUKOCYTES

外源性吗啡类药物和内源性阿片类药物（如B-）的作用 内啡肽，直接调节人体细胞免疫系统 对传染病有潜在的重要影响。都是非法的 药物成瘾和压力诱导的B-内啡肽释放已经被引用 作为增加对传染病易感性的因素， 从普通感冒到肺结核和艾滋病。虽然免疫效果的 免疫细胞暴露于阿片类药物是众所周知的，很少有信息 关于吗啡对免疫功能的作用机制， 细胞在中枢神经系统中，G蛋白偶联阿片类物质家族 存在受体，其解释了多种阿片剂的活性。 这些受体的基因最近已经从两种小鼠中克隆出来。 和人类的来源和它们的结构-一个7-跨膜螺旋 片段蛋白质推导的。然而，当免疫细胞被筛选时， 逆转录酶/聚合酶链反应方法，只有三角洲 Mu受体（莫尔）是一种原发性高表达的受体， 通过配体判断，CNS中的亲和力吗啡结合位点缺失 结合测定和RT-PCR测定表达的莫尔mRNA。事实上唯一 在植物血凝素上发现了高亲和力吗啡结合位点（POR 活化的T淋巴细胞（-）-吗啡可以从这个部位被 （-）-和（+）-吗啡和B-内啡肽的N-末端，但不是通过 DAGO，DADLE和U-50，488。因为这是唯一报道过的， T淋巴细胞上吗啡的位点，重要的是克隆该位点， 确定它是否仅存在于激活的细胞类型上，或者它是否 也存在于未活化的细胞上， 结合测定的阈值。除了可诱导性， 该位点也可能在诱导前组成型表达 并可作为吗啡作用于T细胞的起始步骤 淋巴细胞为了验证这一假设，将通过与 通过RT-PCR或通过表达克隆来检测已知的阿片受体。所述por 然后将使用克隆研究通过以下方法分离的淋巴细胞群： 磁珠/抗体复合基于它们的激活状态。 淋巴细胞的活化状态也将被特异性的 促分裂原刺激这些实验的结果将回答2 免疫药理学重要问题-1。是诱导吗啡 结合位点（POR）足以解释吗啡对 细胞免疫系统？和2.表达与表达之间的相关性是什么 T细胞的诱导结合位点和活化状态 亚型？最终，为了控制鸦片对免疫系统的影响， 系统，有必要了解阿片类药物 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机制的了解， 实验将关闭。

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.