ROLE OF CALCIUM-BINDING PROTEIN AND FUNCTION IN LUNG DISEASE

这个子项目是许多研究子项目中的一个 由NIH/NCRR资助的中心赠款提供的资源。子项目及 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 成功的呼吸道病原体必须能够对哺乳动物肺中广泛的复杂防御机制迅速作出反应。在组织胞浆菌病中，巨噬细胞-下呼吸道的第一道防线-被荚膜组织胞浆菌有效地寄生。这一过程取决于毒力因子的产生，因为这种“二型”真菌经历了温度触发的转换，从一个寄生霉菌形式的寄生酵母形式。一种这样的分子是钙结合蛋白（CBP），其优先由酵母形式分泌并且对于组织胞浆菌毒力是必需的。解开CBP结构和功能的实验在很大程度上依赖于我们开发的端粒穿梭质粒，该质粒已用于互补克隆，基因破坏，RNA干扰和报告基因构建。此外，随机插入突变和微阵列转录谱分析有助于我们识别和表征其他参与组织胞浆菌病发病机制的基因。 鼠疫耶尔森氏菌还显示出两种温度调节的生活方式，这取决于它是在跳蚤还是哺乳动物宿主中定居。人类吸入会导致一种快速和压倒性的疾病，我们正试图通过研究在肺部定植早期阶段被激活的基因来了解肺鼠疫的发展。我们还在继续研究百日咳杆菌的毒力因子之一：气管细胞毒素（TCT）是一种释放的肽聚糖片段，在百日咳中触发气道损伤至关重要。TCT与肽聚糖识别蛋白（PGRP）家族成员的特异性受体结合，我们目前的工作旨在了解宿主反应，包括上皮防御，细胞病理学和重塑。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Successful respiratory pathogens must be able to respond swiftly to a wide array of sophisticated defense mechanisms in the mammalian lung. In histoplasmosis, macrophages -- a first line of defense in the lower respiratory tract -- are effectively parasitized by Histoplasma capsulatum. This process depends on virulence factors produced as this "dimorphic" fungus undergoes a temperature-triggered conversion from a saprophytic mold form to a parasitic yeast form. One such molecule is a calcium-binding protein (CBP) that is secreted preferentially by the yeast form and is essential for Histoplasma virulence. The experiments to unravel CBP structure and function have relied heavily on our development of a telomeric shuttle plasmid that has been used for complementation cloning, gene disruptions, RNA interference, and reporter gene constructs. In addition, random insertional mutagenesis and transcriptional profiling with microarrays are helping us identify and characterize other genes involved in the pathogenesis of histoplasmosis. Yersinia pestis also displays two temperature-regulated lifestyles, depending on whether it is colonizing a flea or mammalian host. Inhalation by humans leads to a rapid and overwhelming disease, and we are trying to understand the development of pneumonic plague by studying genes that are activated during the early stages of pulmonary colonization. We are also continuing studies of one of the virulence factors of Bordetella pertussis: tracheal cytotoxin (TCT) is a released fragment of peptidoglycan that is essential for triggering airway damage in whooping cough. TCT binds to a specific receptor that is a member of the peptidoglycan recognition protein (PGRP) family, and our current work is aimed at understanding host responses that include epithelial defense, cytopathology, and remodeling.