Huang Lu, Kushner Nicole L, Theriault Monique E, Pisu Davide, Tan Shumin, McNamara Case W, Petrassi H Mike, Russell David G, Brown Amanda C
Department of Microbiology and Immunology, College of Veterinary Medicine, Cornell University, Ithaca, NY, United States.
Department of Molecular Biology and Microbiology, Tufts University School of Medicine, Boston, MA, United States.
Front Cell Infect Microbiol. 2018 Aug 14;8:275. doi: 10.3389/fcimb.2018.00275. eCollection 2018.
(Mtb) continues to be a threat to Global Public Health, and its control will require an array of therapeutic strategies. It has been appreciated that high-throughput screens using cell-based assays to identify compounds targeting Mtb within macrophages represent a valuable tool for drug discovery. However, the host immune environment, in the form of lymphocytes and cytokines, is completely absent in a chemical screening platform based on infected macrophages alone. The absence of these players unnecessarily limits the breadth of novel host target pathways to be interrogated. In this study, we detail a new drug screening platform based on dissociated murine TB granulomas, named the Deconstructed Granuloma (DGr), that utilizes fluorescent Mtb reporter strains screened in the host immune environment of the infection site. The platform has been used to screen a collection of known drug candidates. Data from a representative 384-well plate containing known anti-bacterial compounds are shown, illustrating the robustness of the screening platform. The novel deconstructed granuloma platform represents an accessible, sensitive and robust high-throughput screen suitable for the inclusive interrogation of immune targets for Host-Directed Therapeutics.
结核分枝杆菌（Mtb）仍然是全球公共卫生的一大威胁，对其进行控制需要一系列治疗策略。人们已经认识到，利用基于细胞的检测方法进行高通量筛选，以识别巨噬细胞内靶向Mtb的化合物，是药物发现的一种有价值的工具。然而，在仅基于感染巨噬细胞的化学筛选平台中，完全不存在以淋巴细胞和细胞因子形式存在的宿主免疫环境。这些因素的缺失不必要地限制了有待探究的新型宿主靶标途径的广度。在本研究中，我们详细介绍了一种基于解离的小鼠结核肉芽肿的新药物筛选平台，名为解构肉芽肿（DGr），该平台利用在感染部位的宿主免疫环境中筛选的荧光Mtb报告菌株。该平台已用于筛选一系列已知的候选药物。展示了来自一个包含已知抗菌化合物的代表性384孔板的数据，说明了筛选平台的稳健性。这种新型的解构肉芽肿平台是一种可及、灵敏且稳健的高通量筛选方法，适用于全面探究宿主导向治疗的免疫靶标。
