Chemical Probes for Bacteria-Macrophage Interactions

项目摘要 细菌病原体使用不同的策略来逃避宿主的防御和殖民 哺乳动物组织一些细菌逃避宿主免疫系统的某些方面，例如 嗜中性粒细胞和抗体，通过隐藏在巨噬细胞内。然而，宏观经济， 本身就是很好的细菌杀手。因此，细菌如何抵御 巨噬细胞的防御和获得足够的营养生存，甚至复制 仍然是感染生物学中的一个关键问题。我们打算通过以下方式缩小这一知识差距： 开发新的工具来探测巨噬细胞与细菌的相互作用。其中一些工具 可以成为抗菌治疗的基础。我们研究革兰氏阴性菌 细胞内病原体肠道沙门氏菌（Salmonella enterica），其在小鼠体内的巨噬细胞中定殖 伤寒模型我们最近开发了一种定量的，表型的，高含量的，在- 巨噬细胞筛选小分子，防止或增加细菌在内部生长 巨噬细胞根据我们的试点屏幕，我们预计将确定总共 大约118个小分子阻止和5个促进S. enterica 在巨噬细胞内复制。我们建议分析化合物的化学结构， 评价纯度，确定有效浓度50并表征效果 的化合物对感染动力学和细胞形态学使用体内成像。

PROJECT SUMMARY Bacterial pathogens use diverse strategies to evade host defenses and colonize mammalian tissues. Some bacteria evade aspects of the host immune system, such as neutrophils and antibodies, by hiding inside of macrophages. Macrophages, however, are good bacterial killers in their own right. Therefore, how bacteria withstand macrophage defenses and acquire sufficient nutrients to survive and even replicate remains a key question in infection biology. We intend to close this knowledge gap by developing new tools to probe macrophage-bacterial interactions. Some of these tools could become the basis of antibacterial treatments. We study the Gram-negative intracellular pathogen Salmonella enterica, which colonizes macrophages in a murine typhoid fever model. We recently developed a quantitative, phenotypic, high content, in- macrophage screen for small molecules that prevent or increase bacterial growth inside of macrophages. Based on our pilot screen, we anticipate identifying a total of approximately 118 small molecules that prevent and 5 that promote S. enterica replication within macrophages. We propose to analyze compound chemical structure, evaluate purity, and establish the Effective Concentration50 and characterize the effect of compounds on infection dynamics and cell morphology using in vivo imaging.