NTHi Type IV pili: expression and vaccine potential

NTHi IV 型菌毛：表达和疫苗潜力

• 批准号: 7659151
• 负责人: ROBERT S. MUNSON
• 金额: $ 43.78万
• 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/7659151
• 关键词: Adult; Amino Acid Sequence; Antibodies; Antigens; Bacterial Transformation; Binding Sites; Biogenesis; Cell membrane; Cell surface; Child; Chinchilla (genus); Clinical; Competence; Complementary DNA; DNA; Data; Development; Disease; Endotoxins; Epitope Mapping; Fimbriae Proteins; Gene Cluster; Gene Expression Profile; Gene Expression Profiling; Genes; Genetic; Genome; Goals; Gonorrhea; Gram-Positive Bacteria; Haemophilus influenzae; Hemophilus; Immune response; Infection; Knowledge; Laboratories; Lead; Learning; Membrane; Membrane Proteins; Microarray Analysis; Microbial Biofilms; Microbiology; Modeling; Mutagenesis; Nature; Neisseria gonorrhoeae; Nontypable Haemophilus influenza; Organism; Otitis Media; Pasteurellaceae; Pathogenesis; Phenotype; Pilum; Play; Pneumonia; Proteins; Publishing; Regulation; Regulon; Research Personnel; Respiratory Tract Diseases; Role; Streptococcus pneumoniae; Structure; Study Section; Surface; System; Testing; Type II Secretion System Pathway; Vaccines; Vibrio cholerae; Virulence; base; cell motility; design; disorder prevention; in vivo; lipooligosaccharide; mutant; novel strategies; pathogen; periplasm; plasmid DNA; programs; research study; uptake; vaccine development

Nontypeable Haemophilus influenzas (NTHi) is a predominant etiologic agent of otitis media (OM) in chil- dren as well as a major cause of pneumonia and other respiratory tract diseases in adults. Although much has been learned about several outer membrane proteins and the lipooligosaccharide produced by this or- ganism, overall we have a poor understanding of the pathogenesis of Haemop/7/7i;s-induced infection. Type IV pili (Tfp) play a significant role in the pathogenesis of disease caused by numerous Gram-negative pathogens and some Tfp-based immunogens induce protective activity against these organisms. Although a cryptic gene cluster that might encode a Tfp in Haemophilus has long been recognized, ex- pression of Tfp has never been demonstrated by this organism. In fact, in a review entitled DNA uptake dur- ing Bacterial Transformation, published in Nature Reviews Microbiology in March 2004, Chen and Dubnau state "Other competent organisms, such as H. influenzae or the Gram-positive bacteria B. subtilis and S. pneumoniae, require similar genes for DNA uptake, but do not possess filamentous structures that extend from the cell surface". Further, twitching motility, a non-flagellar-based Tfp-dependent motility has never been observed among the Pasteurellaceae. We recently demonstrated that NTHi produces Tfp under defined environmental conditions and further, that Tfp are responsible for twitching motility in NTHi. We hypothesize that Tfp will play an important role in pathogenesis. We have also determined that H. influenzae has a single copy of the pilA gene (encodes the major pilin subunit) and we have now sequenced this gene in 11 NTHi strains. Unlike the situation for N. gonorrhoeae, the derived amino acid sequence of these genes is highly conserved which is a considerable advantage for our vaccine development efforts that intend to target the H. influenzae Tfp. Experiments are thereby proposed herein to determine how the Tfp are assembled as well as how syn- thesis and assembly of these structures are regulated. Additionally, experiments are proposed design a type IV-pilin based immunogen that will induce protection against infection caused by the majority of nontypeable H. influenzae isolates. These experiments will lead to a better understanding of the pathogenesis of NTHi- induced disease as well as new approaches for the prevention of disease caused by NTHi.

不可用的流感嗜血杆菌（NTHI）是CHIL-的主要病因（OM） 成人肺炎和其他呼吸道疾病的主要原因也是主要原因。虽然很多 已经了解了几种外膜蛋白以及该OR-产生的脂肪含糖 总体而言，我们对止血/7/7i; S诱导的感染的发病机理的理解很差。类型IV pili（TFP）在由许多革兰氏阴性病原体引起的疾病的发病机理中起重要作用 一些基于TFP的免疫原对这些生物诱导保护活性。 尽管长期以来已经识别出可能在嗜血杆菌中编码TFP的神秘基因簇，但 这种生物体从未证明TFP的压力。实际上，在评论中，标题为DNA吸收 Ing细菌转化，自然评论于2004年3月，Chen和Dubnau发表了微生物学 国家“其他胜任的生物，例如流感h。 肺炎，需要类似的DNA摄取基因，但不具有延伸的丝状结构 从细胞表面”。此外，抽搐的运动性是一种非鞭毛基于TFP依赖性的运动性 在巴氏菌科中观察到。 我们最近证明，NTHI在定义的环境条件下产生TFP，然后 该TFP负责在NTHI中抽搐运动。我们假设TFP将在 发病。我们还确定，流感h。具有pila基因的单个副本（编码 主要的PILIN亚基），我们现在已经在11个NTHI菌株中对该基因进行了测序。与情况不同 淋病链球菌，这些基因的衍生氨基酸序列是高度保守的，这是一个相当多的 我们的疫苗开发工作的优势，该疫苗开发工作旨在针对流感霍曼氏菌TFP。 因此，本文提出了实验，以确定如何组装TFP以及如何合成 这些结构的论文和组装受到调节。此外，提出了实验设计一种类型 基于IV-pilin的免疫原，将诱导大多数不可替代的感染保护感染 H.流感分离株。这些实验将使人们更好地了解nthi- 诱导疾病以及预防由NTHI引起的疾病的新方法。