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

 描述：药物滥用者占全球艾滋病病例的很大比例。事实上，全世界大约10%的新艾滋病毒感染可归因于注射毒品，通常是海洛因等阿片类药物。HIV C分支病毒是全球近一半的感染病例和印度95%以上感染病例的原因。进化枝C在建立一个不断扩大的流行病的先天能力，可以归因于其独特的生物学特性，再加上药物滥用的共病，这反过来又会影响HIV LTR的遗传多态性。我们最近证明了印度C亚型中出现了4种NF-κB位点启动子变异株。变异病毒株获得获得额外的NF-κB（野生型中3个位点和变异株中4个位点）或RBEIII（野生型中1个位点和变异株中2个位点）结合位点（TFBS）的分子策略，导致病毒启动子的转录效力增强。TFBS复制与过去十年印度变异病毒株的快速扩张有关。事实上，4-κB菌株在印度的流行率从2000-2003年的1-2%增加到十年后的惊人的30-35%。复制NF-κB基序的能力似乎是C亚型所独有的。基于这些观察结果，我们假设C亚型的4-κB菌株更具感染性，导致阿片类药物背景下免疫激活、终末器官发病和潜伏期增加。尚无系统性研究评估病毒变异体生成对疾病进展和/或终末器官受累（特别是CNS）的影响。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。该提案汇集了在分子病毒学，猕猴发病机制和药物滥用方面具有独特优势的研究人员，以回答这些关键问题，同时还解决了进化枝B和C病毒的相关性。具体而言，提出了两个目标。SA1：目的评价在B和C两个进化枝的启动子和env序列中含有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：评估在不存在或存在ART和阿片类药物依赖的情况下感染SHIV克隆（进化枝B vs. C病毒-在SA 1中产生）的印度恒河猴的免疫应答和疾病发病机制。SA 2b：评价在ART和阿片类药物存在下NF-κ B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.