GENETIC REQUIREMENTS FOR THE SURVIVAL OF TUBERCLE BACILLI IN NONHUMAN PRIMATES

这个子项目是许多利用资源的研究子项目之一 由NIH/NCRR资助的中心拨款提供。子项目的主要支持 而子项目的主要调查员可能是由其他来源提供的， 包括其它NIH来源。 列出的子项目总成本可能 代表子项目使用的中心基础设施的估计数量， 而不是由NCRR赠款提供给子项目或子项目工作人员的直接资金。 结核病是由结核分枝杆菌（Mycobacterium tuberculosis，Mtb）引起的一种突发性传染病。每年估计有800万人患结核病，约200万人死于结核病。迫切需要新的药物和疫苗来有效控制结核病。这需要更好地了解结核分枝杆菌如何适应各种各样的环境条件，这些条件是结核分枝杆菌在感染的各个阶段不可避免地面临的。 非人灵长类动物（NHP）可以说是结核病关键方面的最佳模型。因此，分析结核分枝杆菌成功感染NHP肺并在NHP肺中持续存在的机制将非常有用。我们研究了在NHP肺实验暴露于高剂量的结核分枝杆菌转座子突变体的结核分枝杆菌的生长/生存所必需的基因。在该急性TB模型中，所有测试突变体中的33.13%针对体内生长被减毒，而小鼠模型中所有突变体中仅约6%被减毒。在灵长类动物体内存活的减毒Mtb突变体参与脂质毒力因子的运输;细胞壁阿拉伯聚糖和肽聚糖、脂肪酸和聚酮的生物合成; DNA修复;固醇代谢和哺乳动物细胞进入（mce）。我们的研究强调了结核分枝杆菌感染所采用的各种毒力机制，并克服了NHP肺部感染期间遇到的恶劣环境。 我们希望利用我们的能力来模拟人类结核病的各种临床阶段-急性肺结核、慢性进展性结核和NHP中的潜伏性无症状结核-以研究Mtb突变体的生长/存活表型谱。此外，我们想了解的两个Mtb途径的毒力至关重要的作用，使用NHP模型。这些包括mce 1/mce 4操纵子，其成员是在NHP肺中生长减弱的突变体;和dos调节子的成员，令人惊讶的是在NHP肺中没有减弱，尽管它们在潜伏期，持久性和缺氧防御中有明确的作用。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. Tuberculosis (TB), caused by Mycobacterium tuberculosis (Mtb), is an infectious disease emergency. Each year an estimated 8 million people develop, and ~ 2 million people die of TB. New drugs and vaccines are urgently needed to effectively control TB. This requires a better understanding of how Mtb adapts to a wide-variety of environmental conditions, inevitably faced by it during the various stages of infection. Nonhuman Primates (NHPs), arguably, best model critical aspects of TB. Analysis of the mechanisms employed by Mtb to successfully infect and persist in NHP lungs would therefore be very useful. We studied genes essential for growth/survival of Mtb in the NHP lungs experimentally exposed to high doses of Mtb transposon mutants. In this acute model of TB, 33.13% of all tested mutants were attenuated for in-vivo growth compared to the mouse model where only ~6% of all mutants are attenuated. The Mtb mutants attenuated for in-vivo survival in primates were involved in the transport of lipid virulence factors; biosynthesis of cell-wall arabinan and peptidoglycan, fatty-acids and polyketides; DNA repair; sterol metabolism and mammalian cell-entry (mce). Our study highlights the various virulence-mechanisms employed by Mtb for infection and to overcome the hostile environment encountered during infection of NHP lungs. We would like to leverage our ability to model the various clinical phases of human TB - acute, pulmonary TB, chronic-progressive TB and latent, asymptomatic TB in NHPs - to study the growth/survival phenotype profiles of Mtb mutants. Further, we would like to understand the role of two Mtb pathways crucial for virulence, using the NHP model. These include the mce1/mce4 operons, whose members were among mutants that were attenuated for growth in NHP lungs; and members of the dos regulon, which were surprisingly not attenuated in NHP lungs, in-spite of their well-defined roles in latency, persistence and defense against hypoxia.