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

描述（来自申请人摘要）：拟进行研究以进一步定义 阿片激动剂调节宿主-病原体相互作用的能力， 感染某些阿片类药物可以改变感染者体内HIV-1的复制。 细胞阿片类药物也对关键的神经元表达产生显著影响。 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 调节作为关键HIV-1共受体的趋化因子受体。 工作假设是阿片类药物诱导HIV-1复制增加 可以通过天然细胞抗病毒防御机制逆转，即，2 '， 5 '-寡腺苷酸合成酶（2- 5 OAS）/RNase L和p68激酶（PKR）途径。我们 研究将评估阿片类药物对靶点抗病毒途径的影响。 细胞和定义阿片类药物的能力，以调节表达的关键 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α 2-5A衍生物对PBMC的作用， 从HIV-1阳性静脉吸毒者血液中分离的单核细胞 （IVDU），以及正常供体和HIV-1感染的非IVDU。此外，本发明还提供了一种方法， 将进行研究，以确定2-5A衍生物的能力， 抑制HIV-1阳性IVDU中HIV-1原代分离株的复制 病毒分离物来自治疗初治和HAART耐药患者。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)为了确定表达的后果 抗病毒基因2- 5 OAS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- 5 OAS/PKR构建体转导的细胞和单核细胞。 这些研究将采用已建立的细胞系以及原代T细胞， 来源于CD 34+脐带血造血祖干细胞的巨噬细胞。 细胞内免疫将用于确定阿片类药物的作用 对HIV-1复制和细胞因子/细胞因子受体表达的影响 μ、κ和δ-阿片受体的组成型表达。(3)到 确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细胞和单核细胞系中HIV-1储库的再活化 已经用HIV-1 LTR驱动的2- 5 OAS/PKR构建体转导。总的来说， 这些研究应该提供控制HIV-1复制的策略， 病毒的细胞动员/摄取。

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.