NTHi Type IV pili: expression and vaccine potential

NTHi IV 型菌毛：表达和疫苗潜力

• 批准号: 7159334
• 负责人: ROBERT S. MUNSON
• 金额: $ 42.26万
• 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/7159334
• 关键词: Adult; Amino Acid Sequence; Antibodies; Antigens; Appendix; Bacterial Transformation; Binding Sites; Biogenesis; Cell membrane; Cell surface; Child; Chinchilla (genus); Clinical; Competence; Complementary DNA; Condition; DNA; Data; Development; Disease; Endotoxins; Epitope Mapping; Essential Genes; Facility Construction Funding Category; 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; Numbers; 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

DESCRIPTION (provided by applicant): 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, expression of Tfp has never been demonstrated by this organism. In fact, in a review entitled DNA uptake during 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.

描述（由申请人提供）：不可用的流感嗜血杆菌（NTHI）是Children中耳炎（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外膜蛋白以及这种神经植物产生的脂肪糖酸酯，但总的来说，我们对haemop/7/7i; s诱导的感染的发病机理的理解很差。 IV型PILI（TFP）在由众多革兰氏阴性病原体引起的疾病发病机理中起着重要作用，并且某些基于TFP的免疫原子会诱导针对这些生物的保护活性。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的表达。实际上，在细菌转化期间的标题为DNA摄取的综述中，自然评论在2004年3月发表在微生物学上，陈和杜巴瑙国家“其他有能力的生物，例如H. h. th. h. strapity B.b。b.b. utteris和S. pneumoniae，都需要与DNA Uptake的相似基因，但该结构均不超过DNA，但该基因均不可能扩展到该基因构造。此外，在巴氏菌科中从未观察到Twitch运动，一种非鞭毛基于TFP的运动性。我们最近证明，NTHI在定义的环境条件下产生TFP，此外，TFP负责在NTHI中抽搐运动。我们假设TFP将在发病机理中起重要作用。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），现在我们已经在11个NTHI菌株中对该基因进行了测序。与淋病猪笼草的情况不同，这些基因的衍生氨基酸序列是高度保守的，这对于我们的疫苗开发工作是一个可观的优势，该疫苗的开发工作打算瞄准h. the zae tfp。因此，本文提出了实验，以确定如何组装TFP以及如何调节这些结构的合成和组装。此外，提出了实验设计，一种基于IV型纤维蛋白的免疫原，该设计将诱导由大多数不可抑制的H. themane rance株引起的感染的保护。这些实验将使人们更好地了解nthi诱导的疾病的发病机理以及预防由NTHI引起的疾病的新方法。