MOLECULAR MECHANISMS OF OPIOIDS AS COFACTORS IN AIDS

描述：申请者摘要 我们是第一个报道HIV蛋白表达独特的， 免疫调节活动可能是鸡传染性支气管炎的超感染介体 艾滋病的发病机制。接下来我们研究了潜在的分子机制 病毒蛋白在免疫缺乏症发病中的作用 和艾滋病的神经病理并发症。我们也有一个长期的 对静脉注射娱乐性毒品作为辅助因素的作用的兴趣 艾滋病毒感染的易感性和进展。这些利益合并了 在一系列研究中，阿片类药物增强了几个 HIV蛋白介导的免疫致病机制。因此我们的初步研究 结果证明这项提议是成功的。几个实验性的 方法将被用来确定潜在的分子机制 艾滋病毒蛋白和阿片类药物之间的协同作用。人类淋巴细胞将会是 用HIV蛋白+吗啡培养以寻找进一步的增强作用 我们显示的从Th-1到Th-2淋巴细胞优势的转变是 仅由HIV蛋白介导。这将通过检查以下两个方面来实现 定义细胞因子的Th-1和Th-2的基因表达和产生。进一步 我们将确定阿片类药物在体外是否与HIV蛋白协同作用 诱导淋巴细胞或纯化的小胶质细胞和星形胶质细胞的凋亡 以确定阿片类药物和HIV蛋白对大鼠大脑的影响 免疫系统和中枢神经系统。控制实验使用 MU特异性阿片类拮抗剂，纳洛酮和β-呋喃曲拉明，将 确保阿片类药物介导作用的特异性。体内确认 我们的体外中枢神经系统研究将使用我们建立的大鼠模型1 其中HIV蛋白+兴奋性氨基酸激动剂+吗啡 立体定向注射到背侧海马区。脑片将会是 检查大体和微观病理。标记的识别 凋亡以及特定细胞因子基因的表达将是 用直接杂交和一种新的原位聚合酶链式反应检测脑切片 方法。最后，我们将跟进令人振奋的新发现 β-趋化因子及其受体影响易感性和进展 艾滋病毒感染的风险。最近，我们报道了吗啡调节 β-趋化因子及其受体的表达。现在我们要 确定这些效应背后的分子机制。后续 实验将比较和对比来自HIV-和HIV+的临床材料 成人和新生儿以及随后的神经病理学研究将使用 人中枢神经系统组织。阿片类药物作为HIV感染辅助因子的研究 可能会产生治疗阿片成瘾、艾滋病毒感染的新疗法 病人。

DESCRIPTION: Applicant's Abstract We were the first to report that HIV proteins express unique, immunoregulatory activities that may be extra-infectious mediators of the pathogenesis of AIDS. Next we studied the molecular mecnanisms underlying the effects of viral proteins in the development of the immnunodeficiency and neuropathological complications of AIDS. Also we had a long-standing interest in the role of i.v., recreational drugs as co-factors in susceptibility to and progression of HIV infections. These interests merged in a series of studies demonstrating that opioids potentiate several immunopathogenic mechanisms mediated by HIV proteins. Thus our preliminary results support the success of this proposal. Several experimental approaches will be used to determine the molecular mechanism underlying the synergy between HIV proteins and opioids. Human lymphocytes will be cultured with HIV proteins + morphine to look for further potentiation of the shift from Th-1 to Th-2 lymphocyte predominance that we showed was mediated by HIV proteins alone. This will be accomplished by examining both gene expression and production of Th-1 and Th-2 defining cytokines. Further we shall determine whether opioids synergize with HIV proteins in vitro to induce apoptosis of lymphocytes or purified microglia and astrocytes from rat brains to determine the effects of opioids and HIV proteins on the immune system and the CNS respectively. Control experiments using the mu-specific opioid antagonists, naloxone and beta funaltrexamine, will assure the specificity of opioid-mediated effects. In vivo confirmation of our in vitro CNS findings will be undertaken using our established rat mode1 wherein HIV proteins + excitatory amino acid agonists + morphine will be stereotactically injected into the dorsal hippocampus. Brain slices will be examined for gross and microscopic pathology. Identification of markers for apoptosis as well as expression of specific cytokine genes will be determined in brain sections by direct hybridization and a new, in situ, PCR method. Lastly we shall follow-up on the exciting new findings that the beta-chemokines and their receptors affect susceptibility to and progression of HIV infections. Recently, we reported that morphine modulates the expression of the beta-chemokines and their receptors. Now we shall determine the molecular mechanisms underlying these effects. Subsequent experiments will compare and contrast clinical materials from HIV- and HIV+ adults and neonates and the neuropathological studies will subsequently use human CNS tissues. Our studies on opioids as co-factors in HIV infections may yield novel therapies for the treatment of opioid-addicted, HIV-infected patients.