MOLECULAR MECHANISMS OF OPIOIDS AS COFACTORS IN AIDS

描述：申请人摘要 我们是第一个报道HIV蛋白表达独特， 免疫调节活性，可能是外感染介质的 艾滋病的发病机制。 接下来我们研究子其背后分子机制 病毒蛋白在免疫缺陷发展中的作用 和艾滋病的神经病理并发症。 我们也有一个长期的 对静脉注射的作用感兴趣，娱乐性毒品作为 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， HIV蛋白介导的免疫病理机制。 因此，我们的初步 结果表明，这一建议是成功的。 几种实验 方法将用于确定潜在的分子机制， HIV蛋白和阿片类药物之间的协同作用。 人类淋巴细胞将 与HIV蛋白+吗啡一起培养，以寻找进一步增强 我们发现Th-1淋巴细胞优势向Th-2淋巴细胞优势的转变是 由HIV蛋白单独介导。 这将通过检查 Th-1和Th-2定义细胞因子的基因表达和产生。 进一步 我们将确定阿片类药物是否在体外与HIV蛋白协同作用， 诱导淋巴细胞或纯化小胶质细胞和星形胶质细胞凋亡 大鼠大脑，以确定阿片类药物和HIV蛋白对 免疫系统和中枢神经系统。 对照实验使用 μ-特异性阿片类拮抗剂纳洛酮和β-funalcidamine，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结果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+吗啡将被 立体定向注射到背侧海马体。 大脑切片将是 进行大体和显微病理学检查。 标记物的鉴定 细胞凋亡以及特异性细胞因子基因的表达将是 通过直接杂交和一种新的原位PCR在脑切片中测定 法 最后，我们将跟进令人兴奋的新发现， β-趋化因子及其受体影响对 艾滋病毒感染。 最近，我们报道了吗啡调节 β-趋化因子及其受体的表达。 现在我们将 确定这些影响的分子机制。 后续 实验将比较和对比HIV-和HIV+的临床材料 成人和新生儿以及神经病理学研究将随后使用 人CNS组织。 我们对阿片类药物作为HIV感染辅助因子的研究 可能产生新的治疗阿片类药物成瘾，艾滋病毒感染， 患者

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.