Core E: Viral Pathogenesis & Persistence

项目总结/摘要-病毒致病和持久性核心E 核心E支持CFAR研究人员的需求及其创新 项目，同时保持强大的能力，以协助研究人员更传统的基础和翻译 病毒学研究核心利用来自凯斯和皮特的病毒学家的综合实力， RUSTBELT CFAR研究人员对HIV-1发病机制和持续性的研究。核心E提供 研究人员提供创新服务，通过鉴定和量化潜伏的， 表达和诱导的HIV-1，与开创性的离体方法配对，以检查新的潜伏期的功效。 修复剂（LRA）和完全自体系统中的其他治疗策略，进行更深入的表征 HIV-1病毒库动力学和克隆性的研究。凯斯和皮特也有互补的系统， 量化HIV-1持久性，最初开创了超灵敏单拷贝定量PCR（qPCR） 靶向HIV-1 gag/pol的检测，并建立了通过以下方法测量HIV-1 env RNA转录物的新技术： 基于诱导转录的测序。这些方法不断完善和扩展到现在 定量完整与有缺陷的前病毒DNA，并自动灵敏地检测血浆或培养物中的HIV-1 RNA 上清液。核心E还提供先进的病毒生长检测，以评估诱导型，感染性HIV-1， 新的方法，如使用完全自体系统，其性能特性达到或超过黄金 在外地的标准。自动化的进步也提高了生产的产量、一致性和质量， 近全长HIV-1测序，而新技术已经使整合位点分析从相同的 在单个细胞水平上取样。核心E还提供先进的成像平台，以研究HIV-1感染， 病毒体分辨率，包括实时活细胞成像。核心E的重点是尖端病毒学的设计 补充我们的第二个实验室核心，核心F（系统生物学和生物统计学），重点是 通过提供单宿主细胞基因组学和免疫学。转录组学和 Core F的蛋白质组学能力，以及Core E提供的经验证的HIV潜伏期和逆转模型 打开了将这些详细的分析技术应用于基础病毒学前沿研究的大门， 预防、治疗和治愈研究。核心E还将与下列机构协调，支持艾滋病毒预防举措： 临床科学核心D和EHE SWG 2。核心小组将继续与乌干达核心小组密切合作， C，现在是乌干达两个国家艾滋病毒耐药性测试中心之一， 技术，并提高乌干达调查人员的能力 进行信息丰富的病毒学研究，包括治愈研究。最后，也是最重要的是，Core E将 致力于协助CFAR调查人员为其项目获取关键数据。

PROJECT SUMMARY/ABSTRACT –VIRAL PATHOGENESIS & PERSISTENCE CORE E Viral Pathogenesis & Persistence Core E supports the needs of CFAR investigators and their innovative projects, while maintaining strong capacities to assist researchers with more traditional basic and translational virology research. The Core leverages the combined strength of virologists from both Case and Pitt to facilitate research on HIV-1 pathogenesis and persistence by RUSTBELT CFAR investigators. Core E provides investigators with innovative services to measure HIV-1 persistence with assays that identify and quantify latent, expressed, and inducible HIV-1, paired with pioneering ex vivo methods to examine the efficacy of novel Latency Reversal Agents (LRA) and other curative strategies in fully autologous systems, making deeper characterization of HIV-1 reservoir dynamics and clonality widely available. Case and Pitt also have complementary systems to quantify HIV-1 persistence, having originally pioneered ultrasensitive single copy quantitative PCR (qPCR) assays targeting HIV-1 gag/pol and established novel techniques to measure HIV-1 env RNA transcripts by induced transcription–based sequencing. These methods have been continually refined and expanded to now quantify intact vs. defective proviral DNA and automated for sensitive detection of HIV-1 RNA in plasma or culture supernatants. Core E also provides advanced viral outgrowth assays to assess inducible, infectious HIV-1 in new ways, such as using a fully autologous system whose performance characteristics meet or exceed the gold standard in the field. Advances in automation have also increased the throughput, consistency, and quality of near full-length HIV-1 sequencing, while new technologies have enabled integration site analysis from the same sample at a single cell level. Core E also provides advanced imaging platforms to study HIV-1 infection at single virion resolution, including real-time live cell imaging. The focus of Core E on cutting edge virology is designed to complement our second laboratory core, Core F (Systems Biology & Biostatistics), which focuses on the host through offerings in single host cell genomics and immunology. The combination of the transcriptomic and proteomic capabilities of Core F with the validated HIV latency and reversal models made available by Core E opens the door to applying these detailed analytical techniques to cutting edge studies of basic virology, prevention, treatment, and cure research. Core E will also support HIV prevention initiatives in coordination with the Clinical Sciences Core D, and the EHE SWG2. The Core will continue to work closely with Uganda Core C, which is now one of two national sites for HIV drug resistance testing in Uganda, to transfer advanced technology to our partner organizations in Kampala, Uganda, and enhance the ability of Ugandan investigators to conduct informative virological studies including Cure research. Finally, and most importantly, Core E will dedicate its effort toward assisting CFAR investigators to obtain critical data for their projects.