NTHi Type IV pili: expression and vaccine potential

NTHi IV 型菌毛：表达和疫苗潜力

基本信息

批准号：
7728258
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
项目类别：
财政年份：
2005
资助国家：
美国
起止时间：
2005-12-16 至 2011-11-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7728258
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

项目摘要

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.

不可分型流感嗜血杆菌（NTHi）是儿童中耳炎（OM）的主要病原体。德伦病也是成人肺炎和其他呼吸道疾病的主要原因。虽然很多已经了解了几种外膜蛋白和由这种蛋白产生的脂寡糖或- 总的来说，我们对 Haemop/7/7i;s 引起的感染的发病机制了解甚少。 IV型菌毛（Tfp）在多种革兰氏阴性病原体引起的疾病的发病机制中发挥着重要作用一些基于 Tfp 的免疫原可诱导针对这些生物体的保护活性。尽管早已认识到嗜血杆菌中可能编码 Tfp 的神秘基因簇，但该生物体从未证明过 Tfp 的抑制。事实上，在一篇题为 DNA uptake du- ing Bacterial Transformation，2004 年 3 月发表在 Nature Reviews Microbiology，Chen 和 Dubnau 说明“其他有活性的生物体，例如流感嗜血杆菌或革兰氏阳性菌枯草芽孢杆菌和链球菌。肺炎链球菌需要相似的基因来摄取 DNA，但不具有延伸的丝状结构 ”。此外，抽搐运动，一种基于非鞭毛的 Tfp 依赖性运动，从未被在巴斯德氏菌科中观察到。我们最近证明 NTHi 在规定的环境条件下产生 Tfp，并且进一步， Tfp 负责 NTHi 的抽搐运动。我们假设 Tfp 将在以下方面发挥重要作用：发病。我们还确定流感嗜血杆菌具有 pilA 基因的单个拷贝（编码主要菌毛蛋白亚基），我们现在已经在 11 个 NTHi 菌株中对该基因进行了测序。与情况不同的是在淋病奈瑟菌中，这些基因的衍生氨基酸序列高度保守，这是相当大的。这对于我们针对流感嗜血杆菌 Tfp 的疫苗开发工作来说是有利的。因此本文提出实验来确定 Tfp 是如何组装的以及如何同步的这些结构的论文和组装受到监管。此外，实验建议设计一种类型基于 IV-菌毛蛋白的免疫原将诱导针对大多数不可分型引起的感染的保护流感嗜血杆菌分离株。这些实验将有助于更好地了解 NTHi- 的发病机制。诱发疾病以及预防 NTHi 引起的疾病的新方法。