Bordetella cell surface modification and pathogenesis

博德特氏菌细胞表面修饰和发病机制

• 批准号: 10117511
• 负责人: RAJENDAR K DEORA
• 金额: $ 25.33万
• 依托单位: OHIO STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-12-04 至 2022-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10117511
• 关键词: Acellular Vaccines; Active Immunization; Adherence; Affect; Alanine; Amino Acids; Anabolism; Animal Model; Antimicrobial Resistance; Bacteria; Biochemical; Biological; Biological Assay; Blood Circulation; Bordetella; Bordetella pertussis; Cell Fractionation; Cell Line; Cell surface; Cells; Centers for Disease Control and Prevention (U.S.); Chloride Anion Exchanger; Communities; Country; Data; Development; Disease; Edetic Acid; Emerging Communicable Diseases; Enzymes; Epidemic; Epithelial Cells; Fractionation; Generations; Genes; Genomics; Glycine; Glycylglycine; Gram-Negative Bacteria; Gram-Positive Bacteria; Human; Immune; Immunity; In Vitro; Infection; Laboratories; Link; Mass Spectrum Analysis; Membrane; Microbial Biofilms; Modification; Mus; Nasopharynx; National Institute of Allergy and Infectious Disease; Organ; Outcomes Research; Pathogenesis; Pathogenicity; Pathway interactions; Peptide Hydrolases; Peptidoglycan; Pertussis; Pertussis Vaccine; Polysaccharides; Property; Proteins; Public Health; Publishing; Reagent; Research; Resistance; Resolution; Respiratory System; Respiratory Tract Infections; Role; Structure of respiratory epithelium; Surface; System; Techniques; Technology; Teichoic Acids; Testing; Therapeutic; Vaccination; Vaccines; Vibrio cholerae; Virulence; Work; antimicrobial peptide; insight; mouse model; mutant; neutrophil; new therapeutic target; next generation; novel; pathogen; pathogenic bacteria; prevent; respiratory; tool; transmission process; vaccination outcome

The Gram-negative bacterium Bordetella pertussis is a human-restricted pathogen that causes severe respiratory infections and the disease whooping cough or pertussis. Despite high levels and widespread vaccination, there has been a world-wide rise in B. pertussis infections and pertussis. This resurgence is attributed to the ineffectiveness of the current acellular pertussis vaccines to control bacterial numbers in the nasopharynx, bacterial transmission and generation of shorter-lived immunity. How B. pertussis survives effectively in the respiratory tract and circulates in the community is poorly understood. In order to control the resurgence of pertussis, it is critical to discover new strategies and identify new pathogenesis-associated loci and factors. The draABCD locus identified in our laboratory promotes in vitro (i) the resistance to killing by anti-microbial peptides and proteins (AMPs); (ii) polymorphonuclear leukocytes and (iii) the formation of biofilms. We hypothesize that dra is critical for survival of B. pertussis in the mouse respiratory tract. In Specific aim 1, we will utilize well-established mouse models of intranasal infection to examine its role in colonization and biofilm formation in the respiratory tract organs. We will also test whether dra promotes adherence to respiratory epithelium by using epithelial cells lines and respiratory explants. The Dra proteins catalyze the addition of D-alanine in an outer membrane-associated protease-susceptible component. This mechanism of surface modification is distinct from the polysaccharide modification carried out by other known bacterial loci. The Dra-modified surface factor (DMF) is unknown. In Specific Aim 2, using fractionation techniques and mass spectrometry, we will identify the DMF. This work will uncover new virulence strategies that will inform on the reasons of the resurgence of pertussis. Studies of the dra locus and discovering the identity of the DMF will provide important biochemical and biological insights resulting in advanced understanding of surface alterations in B. pertussis. Outcomes from this research will result in targeting of the dra locus and DMF for developing better vaccines and alternative therapeutics.

革兰氏阴性细菌百日咳是一种人限制的病原体，会导致严重 呼吸道感染和疾病百日咳或百日咳。尽管高水平和广泛 疫苗接种，百日咳芽孢杆菌感染和百日咳一直在全球范围内增加。这种复兴是 归因于当前的细胞百日咳疫苗的无效性控制细菌数量 鼻咽，细菌传播和寿命较短​​的免疫力的产生。 B.百日咳如何幸存 在呼吸道中有效地理解了社区的循环。为了控制 百日咳的复兴，发现新策略并确定与发病机理相关的基因座至关重要 和因素。 在我们的实验室中鉴定的DRAABCD基因座促进体外（i）抗微生物杀死的抵抗力 肽和蛋白质（AMPS）； （ii）多形核白细胞和（iii）生物膜的形成。我们 假设DRA对于小鼠呼吸道中百日咳生存至关重要。在特定目标1中，我们将 利用鼻内感染的良好小鼠模型检查其在定殖和生物膜中的作用 呼吸道器官中的形成。我们还将测试DRA是否促进遵守呼吸道 上皮细胞线和呼吸外植体上皮。 DRA蛋白会催化在外膜相关蛋白酶敏感的外部添加D-丙氨酸 成分。这种表面修饰机制与进行多糖修饰不同 由其他已知的细菌基因座。 DRA修饰的表面因子（DMF）尚不清楚。在特定的目标2中，使用 分馏技术和质谱法，我们将确定DMF。 这项工作将揭示新的毒力策略，以告知百日咳复兴的原因。 DRA基因座并发现DMF的身份的研究将提供重要的生化和生物学 洞察力导致对百日咳芽孢杆菌的表面变化有了深入的了解。这项研究的结果 将导致DRA基因座和DMF的靶向，以开发更好的疫苗和替代性疗法。