Core E: Viral Pathogenesis & Persistence

项目摘要/摘要-病毒致病机制和持久核心E 病毒致病机制和持久性核心E支持CFAR研究人员及其创新的需求 项目，同时保持强大的能力，帮助研究人员进行更传统的基本和翻译 病毒学研究。核心利用凯斯和皮特的病毒学家的综合力量来促进 锈带CFAR研究人员对HIV-1致病机理和持久性的研究。酷睿E提供 调查人员提供创新服务，通过识别和量化潜伏期的化验来衡量艾滋病毒-1的持久性， 表达的、可诱导的HIV-1，与开创性的体外方法配对，以检验新潜伏期的有效性 完全自体系统中的逆转药物(LRA)和其他治疗策略，使其具有更深层次的特征 关于HIV-1储存库动力学和克隆性的研究广泛可用。凯斯和皮特也有互补的系统来 量化HIV-1的持久性，最初开创了超灵敏的单拷贝定量聚合酶链式反应(QPCR) 针对HIV-1 Gag/Poll的检测和建立的测量HIV-1包膜RNA转录本的新技术 诱导转录测序。这些方法一直在不断地改进和扩展到现在。 定量完整和缺陷前病毒DNA，并自动灵敏地检测血浆或培养物中的HIV-1RNA 上清液。Core E还提供先进的病毒生长分析，以评估可诱导的、传染性的HIV-1在 新方法，例如使用性能特性达到或超过金牌的完全自体系统 这是这一领域的标准。自动化方面的进步也提高了吞吐量、一致性和质量 接近全长的HIV-1测序，而新技术已经使整合位点分析成为可能 在单个细胞水平上进行采样。Core E还提供先进的成像平台，用于研究单个 病毒粒子分辨率，包括实时活细胞成像。核心E在尖端病毒学方面的焦点被设计 为了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)，它专注于 通过提供单一宿主细胞基因组学和免疫学来进行宿主。转录和转录的结合 Core F的蛋白质组能力与Core E提供的验证的HIV潜伏期和逆转模型 为将这些详细的分析技术应用于基础病毒学的尖端研究打开了大门， 预防、治疗和治愈研究。核心E还将与以下机构协调支持艾滋病毒预防举措 临床科学核心D和EHE SWG2。CORE将继续与乌干达CORE密切合作 C现在是乌干达艾滋病毒耐药性检测的两个国家地点之一，将先进的 向我们在乌干达坎帕拉的伙伴组织提供技术，并增强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，核心E将 致力于协助CFAR调查人员为他们的项目获取关键数据。

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.