Prophage-associated virulence factors in pathogenic Neisseria species

致病性奈瑟菌属中与原噬菌体相关的毒力因子

• 批准号: 7531795
• 负责人: Anna C. Karls
• 金额: $ 7.38万
• 依托单位: UNIVERSITY OF GEORGIA
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-12-01 至 2011-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7531795
• 关键词: Acute suppurative arthritis due to bacteria; Affect; Bacteria; Bacteriophages; Biological Assay; Blood; Blood - brain barrier anatomy; Blood Circulation; Brain; Case Study; Cell membrane; Chromosomes; Cyclophosphamide; DNA; DNA Insertion Elements; Development; Disease; Disease Progression; Elements; Endocarditis; Endothelial Cells; Epithelial; Epithelium; Excision; Exhibits; Genes; Genetic Recombination; Genital system; Genome; Goals; Gonorrhea; Growth; Homologous Gene; Human; Incidence; Individual; Infection; Inflammation; Inflammatory Response; Investigation; Lysogeny; Measures; Mediating; Meningitis; Meningococcal Infections; Mental Retardation; Morbidity - disease rate; Movement; Mucous Membrane; Neisseria; Neisseria gonorrhoeae; Neisseria meningitidis; Pathogenesis; Pelvic Inflammatory Disease; Penetration; Play; Process; Prophages; Protein Secretion; Proteins; Public Health; Qualifying; Quantitative Reverse Transcriptase PCR; Regulation; Replication Initiation; Research; Role; Sepsis; Septic Shock; Septicemia; Serogroup B Neisseria meningitidis; Signal Transduction; Site; Specificity; Staging; Stream; Surface Antigens; Survivors; System; Testing; Tight Junctions; Toxin; Transposase; Vaccines; Vibrio cholerae; Virulence; Virulence Factors; Work; Zot protein; antimicrobial; chemotherapy; deafness; design; experience; mortality; pathogen; response; therapeutic vaccine

DESCRIPTION (provided by applicant): The long-term goal of this research is to identify virulence factors that may be targeted for the design of effective vaccines and therapeutics for Neisseria gonorrhoeae (gonococci, GC), which causes gonorrhoeae and pelvic inflammatory disease, and Neisseria meningitidis (meningococci, MC), which causes septicemia and meningitis. The objective of this R03 application is to examine the role of the meningococcal disease-associated (MDA) prophage in the pathogenesis of GC and MC. The MDA prophage, which is found at multiple loci on the GC and MC genomes, encodes a potential toxin that is a homologue of the Vibrio cholera CTX prophage zonular occulens toxin (Zot) that alters epithelial and endothelial tight junctions. Thus, the MDA prophage Zot proteins in GC and MC have the potential to be involved in inflammation, sepsis, and/or invasion of the blood-brain barrier. The insertion (and excision) of the MDA phage DNA into the host chromosome to establish lysogeny and possible excision to initiation replication may be mediated by the transposase of the ISNgo2/3 elements, which are part of the phage genome. Defining the mechanism for the movement of the phage DNA is essential to understanding the role of the phage in GC and MC pathogenesis. The specific aims are to: 1. Determine the activities of the meningococcal and gonococcal Zot proteins, and 2. Define the role of the ISNgo2/3 transposase, Irg, in directing insertion and/or excision of MDAphi DNA and controlling zot expression in MC and GC. Experimental approaches will include assays for zonular occludens toxin activity for Zot from MC and GC on epithelial and brain microvascular endothelial cell membranes; quantitative reverse transcriptase PCR to measure expression of zot and irg under different growth conditions; and quantitative PCR to detect movement and replication of MDAphi. This work is relevant to public health because there is no effective vaccine for Neisseria gonnorrhoeae or for the most prevalent serogroup (B) of Neisseria meningitidis in the U.S. The annual U.S. incidence of gonococcal disease is 130 in 100,000, while the incidence of meningococcal disease is 1 in 100,000. Despite the use of chemotherapeutics, 10-40% of invasive meningococcal infections are fatal and 10-15% of survivors have serious sequelae, including mental retardation and deafness.

描述(由申请人提供)： 这项研究的长期目标是确定可能针对淋球菌(淋球菌，GC)和脑膜炎奈瑟菌(脑膜炎球菌，MC)的有效疫苗和治疗方法的毒力因素。淋球菌引起淋病和盆腔炎，脑膜炎奈瑟菌引起败血症和脑膜炎。此R03应用程序的目的是检查脑膜炎双球菌疾病相关(MDA)原噬菌体在GC和MC发病机制中的作用。丙二醛原噬菌体在GC和MC基因组的多个位点上发现，它编码一种潜在的毒素，它是霍乱弧菌CTX原噬菌体透明带毒素(Zot)的同源物，可以改变上皮和内皮的紧密连接。因此，GC和MC中的丙二醛原噬菌体Zot蛋白有可能参与炎症、败血症和/或血脑屏障的入侵。丙二醛噬菌体DNA插入(和切除)到宿主染色体中以建立溶原性和可能的切除以启动复制可能是由作为噬菌体基因组一部分的ISNgo2/3元件的转座酶介导的。明确噬菌体DNA的运动机制对于理解噬菌体在GC和MC发病机制中的作用是至关重要的。其具体目的是：1.确定脑膜炎球菌和淋球菌Zot蛋白的活性；2.确定ISNgo2/3转座酶IRG在指导MDAphi DNA插入和/或切除以及控制MC和GC中Zot表达中的作用。实验方法将包括MC和GC对Zot在上皮和脑微血管内皮细胞膜上的Zot活性的检测；定量逆转录聚合酶链式反应(QRT)检测不同生长条件下ZOT和IRG的表达；以及定量聚合酶链式反应(QTCR)检测MDAphi的运动和复制。 这项工作与公共卫生有关，因为目前还没有针对淋球菌或美国最流行的脑膜炎奈瑟菌血清群(B)的有效疫苗。美国淋球菌疾病的年发病率为每10万人中有130人，而脑膜炎球菌疾病的发病率为每10万人中有1人。尽管使用了化疗药物，10%-40%的侵袭性脑膜炎双球菌感染是致命的，10%-15%的幸存者有严重的后遗症，包括智力低下和耳聋。