Effect of small molecule inhibitors on transcriptome of B6.Sst1S bone marrow derived macrophages stimulated with TNF and infected with virulent Mycobacterium tuberculosis

We investigated candidate host-directed therapies for tuberculosis in vitro. BMDMs from tuberculosis susceptible mice B6.Sst1S were stimulated with TNF and infected with Mtb. The infected macrophage were treated with Rocaglamide A, ISRIB and JNK inhibitor SP600125 alone or in combinations to asses the inhibitors' effectas on macrophage transcriptome. Overall design: B6.Ssts1S BMDMs were stimulated with 10 ng/ml of TNF for 16h and infected with Mtb H37Rv at MOI=1. At 72h post infection total RNA was isolated and processed for RNA-seq

本研究体外探究了针对结核病的候选宿主导向治疗方案。以结核分枝杆菌易感小鼠B6.Sst1S的骨髓源性巨噬细胞（Bone Marrow Derived Macrophages, BMDMs）为实验材料，经肿瘤坏死因子（Tumor Necrosis Factor, TNF）刺激后感染结核分枝杆菌（Mycobacterium tuberculosis, Mtb）。将感染后的巨噬细胞分别或联合使用罗卡胺A（Rocaglamide A）、ISRIB以及JNK抑制剂SP600125进行处理，以评估上述抑制剂对巨噬细胞转录组的影响。实验整体设计：将B6.Sst1S小鼠的骨髓源性巨噬细胞以10 ng/ml的肿瘤坏死因子刺激16小时，以感染复数（Multiplicity of Infection, MOI）=1的比例感染结核分枝杆菌H37Rv；于感染后72小时提取总RNA，开展RNA测序（RNA-seq）分析。