Acellular vaccines against bacterial pathogens

针对细菌病原体的无细胞疫苗

基本信息

批准号：
7221194
负责人：
Joseph T Barbieri
金额：
$ 32.27万
依托单位：
MEDICAL COLLEGE OF WISCONSIN
依托单位国家：
美国
项目类别：
财政年份：
1990
资助国家：
美国
起止时间：
1990-07-01 至 2010-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7221194
关键词：
ADP Ribose Transferases ADP ribosylation Acellular Vaccines Actins Address Amino Acids Antibiotic Resistance Bacterial Infections Biochemical Burn injury Cell Death Cell physiology Cell surface Cultured Cells Cystic Fibrosis Cytoskeleton Cytotoxin Electrostatics Environment Enzymes Eye Generations Genes Goals Growth Hospitals Host Defense Immune response Immune system Individual Infection Infection Control Intoxication Life Style Localized Mammalian Cell Measures Membrane Molecular Mutagenesis Nuclear Numbers Opportunistic Infections Outcome Pathogenesis Pathway interactions Patients Phagocytosis Physiological Property Proteins Pseudomonas aeruginosa Range Role Signal Pathway Signal Transduction Pathway Site Specificity Substrate Specificity System Testing Toxin Virulence Wound Healing Yersinia base extracellular in vivo inhibitor/antagonist insight molecular modeling pathogen prevent rho GTP-Binding Proteins rho GTPase-activating protein tissue regeneration

项目摘要

DESCRIPTION (provided by applicant): Pseudomonas aeruginosa is a ubiquitous Gram-negative opportunistic pathogen of compromised patients. Especially susceptible are individuals inflicted with burn wounds, eye complications, and cystic fibrosis. P. aeruginosa pathogenesis is facilitated by growth in hospital environments and its intrinsic resistant to antibiotics. P. aeruginosa produces a number of virulence determinants, which are either cell-surface components or secreted. P. aeruginosa produces four type-III secreted cytotoxins (ExoS, ExoT, ExoU, and ExoY), which inactivate the host innate immune system, the first line of defense against bacterial infection. The goals of this proposal are to define the molecular properties of the P. aeruginosa type-III cytotoxins ExoS and ExoT. ExoS and ExoT are bi-functional cytotoxins: the amino terminus of ExoS and ExoT inactivate Rho GTPases by acting as RhoGTPase Activating Proteins (RhoGAPs) to disassemble the actin cytoskeleton and inhibit internalization of P. aeruginosa by mammalian cells, while the carboxyl terminus of ExoS and ExoT comprises an ADPribosyltransferase domain. ExoS and ExoT ADP-ribosylate unique subsets of host proteins in mammalian cells. The targets of ExoS and ExoT control numerous cellular processes, including wound healing, tissue regeneration, and phagocytosis. This proposal addresses the basis for the physiological actions of ExoS and ExoT. Utilizing complementary molecular, cellular, and biochemical approaches, the goals of this application are to: characterize the ADP-ribosylation of Crk by ExoT; determine how ExoT and ExoS recognize their substrates for ADP-ribosylation, characterize the in vivo ADP-ribosyltransferase activity of ExoS, characterize the RhoGAP domain of ExoS, and measure the intracellular targeting of type-III cytotoxins in mammalian cells. Addressing these aims will define how ExoS and ExoT allow P. aeruginosa to establish infection in mammalian cells and define new strategies to detect, prevent, and control infection by this opportunistic pathogen.

描述（由申请人提供）：铜绿假单胞菌是受损患者中普遍存在的革兰氏阴性机会致病菌。特别容易受到烧伤、眼部并发症和囊性纤维化的个体的影响。医院环境中的生长及其对抗生素的内在耐药性促进了铜绿假单胞菌的发病机制。铜绿假单胞菌产生许多毒力决定簇，它们要么是细胞表面成分，要么是分泌的。铜绿假单胞菌产生四种 III 型分泌细胞毒素（ExoS、ExoT、ExoU 和 ExoY），它们使宿主先天免疫系统（抵御细菌感染的第一道防线）失活。该提案的目标是定义铜绿假单胞菌 III 型细胞毒素 ExoS 和 ExoT 的分子特性。 ExoS 和 ExoT 是双功能细胞毒素：ExoS 和 ExoT 的氨基末端通过充当 RhoGTP 酶激活蛋白 (RhoGAP) 来灭活 Rho GTP 酶，分解肌动蛋白细胞骨架并抑制哺乳动物细胞对铜绿假单胞菌的内化，而 ExoS 的羧基末端ExoT包含ADPribosyltransferase结构域。 ExoS 和 ExoT ADP 核糖基化哺乳动物细胞中宿主蛋白的独特子集。 ExoS 和 ExoT 的靶标控制着许多细胞过程，包括伤口愈合、组织再生和吞噬作用。该提案阐述了 ExoS 和 ExoT 生理作用的基础。利用互补的分子、细胞和生化方法，本申请的目标是：表征 ExoT 对 Crk 的 ADP-核糖基化；确定 ExoT 和 ExoS 如何识别其 ADP-核糖基化底物，表征 ExoS 的体内 ADP-核糖基转移酶活性，表征 ExoS 的 RhoGAP 结构域，并测量哺乳动物细胞中 III 型细胞毒素的细胞内靶向。解决这些目标将定义 ExoS 和 ExoT 如何允许铜绿假单胞菌在哺乳动物细胞中建立感染，并定义检测、预防和控制这种机会性病原体感染的新策略。