EFFECT OF OPIOIDS ON 2-5OAS/PKR PATHWAY IN HIV INFECTION

描述(来自申请者的摘要)：建议进行研究以进一步定义 阿片类激动剂调节宿主-病原体相互作用的能力 感染。某些阿片类药物作用于改变HIV-1在感染者体内的复制 细胞。阿片类药物也对关键基因的表达产生显著影响 趋化因子和趋化因子受体基因，并具有调节两者的能力 免疫细胞向感染部位的运输，以及 作为关键的HIV-1辅助受体的趋化因子受体的调节。 工作假说是阿片类药物导致HIV-1复制增加 可以被天然的细胞抗病毒防御机制逆转，即2‘， 5‘-寡腺苷合成酶(2-5OAS)/核糖核酸酶L和p68激酶(PKR)通路。我们的 研究将评估阿片类药物对靶标抗病毒途径的影响 并定义阿片类药物调节关键蛋白表达的能力 细胞因子/细胞因子受体。建议：(1)测定抗HIV-1 代谢稳定、无毒的2-5A衍生物对人外周血单核细胞的影响 HIV-1阳性静脉吸毒者血液中单核细胞的分离 (IVDU)，以及正常捐赠者和感染艾滋病毒-1的非IVDU。此外, 将进行研究以确定2-5A衍生物的能力 抑制HIV-1阳性静脉注射吸毒者的HIV-1初级分离株复制 来自治疗初学者和抗HAART患者的病毒分离株。HIV-1 复制水平将与关键细胞因子的表达相关 和趋化因子受体。(2)确定表达的后果 抗病毒基因2-5OAS和PKR对阿片类药物增强T细胞中HIV-1复制的影响 用HIV-1-LTR驱动的2-5OAS/PKR构建载体转导细胞和单核细胞。 这些研究将使用已建立的细胞系以及原代T细胞和 脐血CD34+造血祖干细胞来源的巨噬细胞。 将利用细胞内免疫来确定阿片类药物的影响 HIV-1复制及细胞因子/细胞因子受体表达的研究 Mu、kappa和增量阿片受体的组成性表达。(3)至 测定阿片类药物对细胞因子/细胞因子受体表达的影响 潜伏感染T细胞和单核细胞株中HIV-1储存库的重新激活 通过HIV-1LTR驱动的2-5OAS/PKR构建载体进行转导。总的来说， 这些研究应该提供控制艾滋病毒-1复制和 细胞动员/摄取病毒。

DESCRIPTION (from applicant's abstract): Studies are proposed to further define the capacity of opioid agonists to modulate host-pathogen interactions during infection. Certain opioids act to alter the replication of HIV-1 in infected cells. Opioids also exert a significant impact on expression of critical chemokines and chemokine receptor genes and have the capacity to regulate both the trafficking of immune cells to sites of infection, as well as the regulation of chemokine receptors, which serve as critical HIV-1 co-receptors. The working hypothesis is that the opioid induced increase in HIV-1 replication can be reversed by natural cellular antiviral defense mechanisms, i.e., the 2', 5'-oligoadenylate synthetase (2-5OAS)/RNase L and p68 kinase (PKR) pathway. Our studies will evaluate the impact of opioids on the antiviral pathways of target cells and define the capacity of opioids to modulate the expression of critical cytokines/cytokine receptors. It is proposed: (1) To determine the anti-HIV-1 effects of metabolically stable, non-toxic a 2-5A derivative on PBMC and monocytes isolated from the blood of HIV-1 positive intravenous drug users (IVDU), as well as normal donors and HIV-1-infected non-IVDU. In addition, studies will be conducted to determine the ability of a 2-5A derivative to inhibit replication of primary isolates of HIV-1 from HIV-1-positive IVDU using viral isolates from both therapy-naive and HAART-resistant patients. HIV-1 replication levels will be correlated with the expression of critical cytokines and chemokine receptors. (2) To determine the consequences of expression of the antiviral genes, 2-5OAS and PKR, on opioid-potentiated HIV-1 replication in T cells and monocytes transduced with HIV-1-LTR-driven 2-5OAS/PKR constructs. These studies will employ established cell lines as well as primary T cells and macrophages derived from CD34+ cord blood hematopoietic progenitor stem cells. Intracellular immunization will be utilized to determine the effects of opioids on HIV-1 replication and cytokine/cytokine receptor expression in cells with constitutive expression of mu, kappa, and delta-opioid receptors. (3) To determine the effects of opioids on cytokine/cytokine receptor expression and reactivation of HIV-1 reservoirs in latently infected T cell and monocyte lines that have been transduced with HIV-1 LTR-driven 2-5OAS/PKR constructs. Overall, these studies should provide strategies to control HIV-1 replication and cellular mobilization/uptake of the virus.