The combinatorial effects of Opiates and the emerging promoter-variant strains of HIV-1 subtype C on HIV neuropathogensis and latency

 描述：吸毒者在全球艾滋病病例中占相当大的比例。事实上，全世界约10%的新艾滋病毒感染可归因于注射吸毒，通常是海洛因等鸦片类药物。全球近一半的感染病例和印度95%以上的感染病例都是由艾滋病毒C分支病毒造成的。C分支在建立扩大流行方面的先天能力可归因于其独特的生物学特性，再加上药物滥用的共同发病率，这反过来又可能影响HIV LTR基因的多态。我们最近证明在印度的C亚型中出现了4个NF-κB位点启动子变异株。变异株获得了获得额外的NF-κB(野生型3个，变异株4个)或RBEIII(野生型1个，变异株2个)结合位点(TFBS)的分子策略，从而提高了病毒启动子的转录效力。TFBS的复制与印度在过去十年中变异病毒株的迅速扩张有关。事实上，印度的4-κB型毒株的流行率已经从2000年至2003年的1-2%上升到10年后的惊人的30%-35%。复制NF-κB基序的能力似乎是C亚型所特有的。基于这些观察，我们假设C亚型的4-κB菌株更具传染性，导致免疫激活、终末器官发病和阿片类药物背景下的潜伏期增加。没有系统性研究评估病毒变异的产生对疾病进展和/或终末器官受累的影响，特别是在中枢神经系统。这种纵向研究只能在艾滋病毒感染和阿片类药物滥用的动物模型中进行。这项建议将在分子病毒学、猕猴发病机制和物质滥用方面具有独特优势的研究人员聚集在一起，以回答这些关键问题，同时也解决了B和C分支病毒的相关性。具体地说，提出了两个目标。目的：评价B、C两个分支病毒启动子中含有1、2、3或4个NF-κB结合位点的SHIV克隆的体外复制能力。Sa 2a：在没有或存在ART和阿片类药物依赖的情况下，评估感染SIV克隆(分支B与C病毒-在SA1中产生)的印度恒河猴的免疫反应和疾病发病机制。SA 2b：在ART和阿片类药物存在的情况下，评估核因子-kB数量对病毒潜伏期的影响。

 DESCRIPTION: Drug abusers account for a substantial proportion of AIDS cases globally. In fact, approximately 10% of new HIV infections worldwide are attributable to injecting drug use, often of an opiate such as heroin. HIV clade C viruses are responsible for nearly half of the global infections and more than 95% of the infections in India. The innate ability of clade C in establishing an expanding epidemic could be ascribed to its unique biological properties, coupled with co-morbidity of drug abuse, which in turn, could influence the HIV LTR genetic polymorphism. We have recently demonstrated the emergence of 4 NF-κB site promoter variant strains in the subtype C in India. The variant viral strains acquire the molecular strategy of attaining an additional NF-κB (3 sites in the wild type & 4 in the variant strains) or RBEIII (onein the wild type & 2 in the variant strains) binding sites (TFBS) leading to enhanced transcriptional potency of the viral promoter. The TFBS duplication is associated with rapid expansion of the variant virus strains in India over the past decade. In fact, the prevalence of the 4-κB strains i India has increased from 1-2% during 2000-2003 to a staggering 30-35% a decade later. The ability to duplicate the NF-κB motifs appears to be unique to subtype C. Based on these observations, we hypothesize that 4-κB strains of subtype C are more infectious leading to increased immune activation, end-organ pathogenesis and latency in the context of opiates. No systemic studies have assessed the impact of viral variant generation on disease progression, and/or end-organ involvement specifically in the CNS. Such longitudinal studies can only be possible in an animal model of HIV infection and opiate abuse. This proposal brings together investigators with unique strengths in molecular virology, macaque pathogenesis and substance abuse to answer these crucial questions while also addressing the relevance in the context of clade B and C viruses. Specifically, two aims are proposed. SA1: To assess the in vitro replication competence of SHIV clones containing 1, 2, 3 or 4 NF-κB binding sites in the promoter as well as env sequences of both clades B and C viruses. SA 2a: To assess the immune responses and disease pathogenesis in Indian rhesus macaques infected with the SHIV clones (clade B vs. C viruses - generated in SA1) in the absence or presence of ART & opiate dependence. SA 2b: To evaluate the influence of the NF-kB number on viral latency in the presence of ART and opiates.