Core B: Animal Model Core

共享资源核心B -摘要 该综合P01程序依赖于呼吸道飞沫的生物物理学影响M的前提。 结核病（Mtb）的有效载荷，生理学和可培养性，从而影响其传播和感染的能力。 新宿主研究感染的空气生物学，并确定有效治疗所需的细菌基因， 传播，我们将开发一个易于处理的模拟结核病传播模型，并将其应用于比较传染性 野生型与突变型结核分枝杆菌菌株，从腔酪蛋白终端肺气道。老鼠是世界上 用于研究结核病、细菌遗传要求的广泛的气溶胶感染动物模型 和对治疗的反应有大量的实验证据支持有效的肺“播种”， 雾化的结核分枝杆菌液滴。我们集团有两个大型动物BSL 3设施，并有一个长期的 在开发和优化新的动物模型，以解决具有挑战性的问题， 翻译工具 我们还开发了在改良的活动性结核病兔模型中产生腔酪蛋白的工具， 依赖于泡沫状巨噬细胞裂解物和粘膜唾液分泌模拟物作为起始的酪蛋白替代物 点这些基质将被优化并用于（i）Mtb突变体的体外筛选以鉴定和验证 Mtb在宿主样和环境之间过渡期间生存所需的候选基因 项目1和2：确定结核分枝杆菌需要的基因和适应性代谢反应， 存活的传输应力），（ii）作为起始材料，以雾化结核分枝杆菌在实验中的控制 传输到小鼠，以定性和定量地模拟人呼吸道分泌物的总体特征 生物气溶胶发生器中的动力学，以及（iii）测试某些药物可能损害传播的假设 尽管在干酪和牙腔中以亚杀菌浓度积累。 在空气生物学项目4的支持和指导下，我们将联合收割机并利用体外、离体和体内技术 体内工具开发和优化通过受控气溶胶感染模拟传播的小鼠模型。的 模型将被应用于确认Mtb基因和途径的贡献-体外鉴定-在成功的 从一个主机到另一个主机的多步转换。在小鼠中表现出传播适应性丧失的突变体 将在豚鼠中进行动物间传播试验（项目3：M. 结核病传播）。 因此，核心B活动服务于每个项目的目标，并将受到空气生物学项目4的密切指导。

Shared Resource Core B – Abstract This integrated P01 program relies on the premise that the biophysics of respiratory droplets impacts M. tuberculosis (Mtb) payload, physiology and culturability, and thus influences its ability to transmit and infect a new host. To study the aerobiology of infection and identify the bacterial genes required for effective transmission, we will develop a tractable model of simulated Mtb transmission and apply it to compare infectivity of wild type versus mutant Mtb strains, from cavity caseum to terminal lung airways. The mouse is the most widespread animal model of aerosol infection used to investigate TB disease, bacterial genetic requirements and response to therapy. There is extensive experimental evidence supporting efficient lung ‘seeding’ by aerosolized Mtb droplets. Our group has access to two large animal BSL3 facilities and has a long-standing expertise in developing and optimizing new animal models to address challenging questions that require translational tools. We have also developed tools to generate cavity caseum in a modified rabbit model of active TB, and variations of caseum surrogate that rely on foamy macrophage lysate and mucosalivary secretion mimics as the starting point. These matrices will be optimized and used (i) in in vitro screens of Mtb mutants to identify and validate candidate genes required for survival of Mtb during the transitions between host-like and environmental conditions (with Projects 1 and 2: identification of genes and adaptive metabolic responses that Mtb requires to survive transmission stresses), (ii) as the starting material to aerosolize Mtb bacilli in experiments of controlled transmission to mice, to qualitatively and quantitively mimic overall features of human respiratory secretion dynamics in the bioaerosol generator, and (iii) to test the hypothesis that some drugs may impair transmission despite accumulating in caseum and cavities at concentrations that are sub-bactericidal. With support and guidance from the Aerobiology Project 4, we will combine and leverage in vitro, ex vivo and in vivo tools to develop and optimize a mouse model of simulated transmission by controlled aerosol infection. The model will be applied to confirm the contribution of Mtb genes and pathways – identified in vitro – in the successful multi-step transition from one host to another. Mutants with demonstrated loss of fitness for transmission in mice will progress to testing between-animal transmission in guinea pigs (Project 3: Mechanisms of cough in M. tuberculosis transmission). Thus Core B activities serve the objectives of each project and will be closely guided by Aerobiology Project 4.