NTHi Type IV pili: expression and vaccine potential

NTHi IV 型菌毛：表达和疫苗潜力

• 批准号: 7525064
• 负责人: ROBERT S. MUNSON
• 金额: $ 2.14万
• 依托单位: RESEARCH INST NATIONWIDE CHILDREN'S HOSP
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-12-16 至 2010-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7525064
• 关键词: Adherence; Adult; Amino Acid Sequence; Antibodies; Antigens; Bacterial Transformation; Binding Sites; Biogenesis; Cell membrane; Cell surface; Child; Chinchilla (genus); Clinical; Collaborations; Competence; Complementary DNA; Condition; Crystallization; DNA; Data; Data Set; Development; Disease; Endotoxins; Epitope Mapping; Essential Genes; Exhibits; Facility Construction Funding Category; Fimbriae Proteins; Funding; Future; Gene Cluster; Gene Expression Profile; Gene Expression Profiling; Genes; Genetic; Genome; Goals; Gonorrhea; Gram-Positive Bacteria; Haemophilus influenzae; Hemophilus; Hemophilus influenza infection; Heterogeneity; Immune response; Immunity; Immunodominant Epitopes; Infection; Knowledge; Laboratories; Learning; Membrane; Methods; Microarray Analysis; Microbial Biofilms; Microbiology; Modeling; Mutagenesis; Nature; Neisseria gonorrhoeae; Nontypable Haemophilus influenza; Organism; Otitis Media; Pasteurellaceae; Pathogenesis; Phase; Phenotype; Pilum; Play; Pneumonia; Proteins; Pseudomonas aeruginosa; Publishing; Recombinants; Regulation; Regulon; Request for Applications; Respiratory System; Respiratory Tract Diseases; Role; Selenomethionine; Sequence Homology; Streptococcus pneumoniae; Structure; Study Section; Surface; System; Testing; Type II Secretion System Pathway; Vaccines; Vibrio cholerae; Virulence; base; cell motility; design; expectation; human disease; in vivo; lipooligosaccharide; mutant; pathogen; periplasm; plasmid DNA; protein structure; research study; uptake

DESCRIPTION (provided by applicant): Nontypeable Haemophilus influenzae (NTHi) is a predominant etiologic agent of otitis media (OM) in children as well as a major cause of pneumonia and other respiratory tract diseases in adults. Although much has been learned about several surface- exposed proteins and the endotoxin (lipooligosaccharide) produced by this organism, overall we have a poor understanding of the pathogenesis of Haemophilus-induced infection. Type IV pili (Tfp) play a significant role in the pathogenesis of disease caused by Pseudomonas aeruginosa, Vibrio cholerae, Neisseria gonorrhoeae and numerous other Gram-negative pathogens. Tfp-based immunogens induce protective activity against these organisms, although in some systems, including N. gonorrhoeae, protection against infection induced by these immunogens is limited due to sequence diversity of the pilin protein. We recently demonstrated that Tfp are produced by NTHi. More recently, we demonstrated that these Tfp play a role in adherence and biofilm formation in the chinchilla model of otitis media caused by NTHi. We also demonstrated that the product of the pilA gene, PilA, exhibits limited antigenic heterogeneity suggesting that it might be a useful vaccine candidate. We are currently funded to characterize the genes and gene products required for Tfp biogenesis in NTHi and to epitope map the PilA protein. We have made considerable progress toward these goals. However, as we endeavor to identify both the surface-exposed domains and immunodominant epitopes within the type IV pilus protein, it will be important to have detailed structural information on the NTHi type IV pilus. We established a collaboration with Dr. Lisa Craig to determine conditions for crystallization of the Haemophilus type IV pilin with the expectation of ultimately seeking future supplemental funding to determine the structure. These studies were funded by a modest 2-year subcontract as part of this funded initiative. Dr. Craig's group has succeeded in crystallizing recombinant pilin and has collected a 1.7 Angstrom data set. This competing supplemental application requests an additional 17 months of subcontract funding in order that Dr. Craig's group can complete the structural analysis of the PilA protein. Project Narrative Nontypeable Haemophilus influenzae is an important cause of otitis media in children as well as other diseases of the human respiratory tract. Recently, we developed evidence indicating that a structure on the bacterial surface, called a type IV pilus, was important for infection. Further, our data indicate that the major protein component of the type IV pilus should be considered for inclusion in an otitis media vaccine. Understanding the structure of this protein is critical to understanding the role of this protein in infection and immunity. We have made considerable progress toward this goal and request funds to complete determination of the structure of this protein.

描述（由申请人提供）：不可用的流感嗜血杆菌（NTHI）是儿童中耳炎培养基（OM）的主要病因，也是成人肺炎和其他呼吸道疾病的主要原因。尽管已经了解了几种表面暴露的蛋白质和该生物体产生的内毒素（脂肪糖糖）的知识，但总体而言，我们对嗜血杆菌诱导的感染的发病机理的理解不足。 IV型Pili（TFP）在由铜绿假单胞菌，弧菌霍乱，淋病奈瑟氏菌和许多其他革兰氏阴性病原体引起的疾病发病机理中起重要作用。基于TFP的免疫原诱导了针对这些生物的保护活性，尽管在某些系统中，包括淋病链球菌，防止这些免疫原子引起的感染受到pILIN蛋白的序列多样性的限制。我们最近证明了TFP是由NTHI生产的。最近，我们证明了这些TFP在NTHI引起的中耳炎的龙骨模型中起着粘附和生物膜形成的作用。我们还证明了pila基因PILA的产物表现出有限的抗原异质性，这表明它可能是一种有用的疫苗候选者。我们目前被资助以表征NTHI中TFP生物发生所需的基因和基因产物和表位图绘制pila蛋白。我们已经朝着这些目标取得了长足的进步。但是，随着我们努力确定IV型菌毛蛋白中的表面暴露结构域和免疫主流表位，对NTHI IV型Pilus进行详细的结构信息将很重要。我们与丽莎·克雷格（Lisa Craig）博士建立了一项合作，以确定嗜血杆菌IV型PILIN结晶的条件，并期望最终寻求未来的补充资金来确定结构。这些研究是由适度的2年分包合同资助，作为这项资助的倡议的一部分。 Craig博士的小组成功地结晶了重组PILIN，并收集了1.7 Angstrom数据集。该竞争性补充申请要求另外17个月的分包资金，以便Craig博士的小组可以完成对Pila蛋白的结构分析。项目叙述性不可用的流感嗜血杆菌是儿童中耳炎以及人类呼吸道的其他疾病的重要原因。最近，我们开发了证据，表明细菌表面上的结构（称为IV型菌毛）对于感染很重要。此外，我们的数据表明，应考虑将IV型菌毛的主要蛋白质成分纳入中耳炎培养基疫苗。了解该蛋白质的结构对于理解该蛋白在感染和免疫力中的作用至关重要。我们已经在这一目标方面取得了长足的进步，并要求资金完成该蛋白质结构的确定。