V. cholerae proteins are required for biofilm growth-vaccine targets

生物膜生长疫苗靶标需要霍乱弧菌蛋白

• 批准号: 8142027
• 负责人: WILLIAM Franklin WADE
• 金额: $ 19.27万
• 依托单位: DARTMOUTH COLLEGE
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-09-15 至 2013-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8142027
• 关键词: Adult; Affect; Antibodies; Antigens; Bacteria; Bacterial Antigens; Bile fluid; Binding; Biological Assay; Biology; Biomass; Blood Circulation; Carbohydrates; Carrier Proteins; Cells; Cessation of life; Characteristics; Child; Cholera; Cholera Vaccine; Complex; Development; Disease; Environment; Epitopes; Gene Cluster; Gene Expression Regulation; Generations; Genes; Goals; Growth; Growth and Development function; Homologous Gene; Human; Immune Sera; Immune system; Immunization; Immunoglobulin G; Immunologist; In Vitro; Individual; Infection; Intestines; Investigation; Kinetics; Laboratories; Lethal Dose 50; Microbial Biofilms; Modeling; Molecular; Morphology; Mus; Operon; Oxidative Stress; Pathogenesis; Phenotype; Polysaccharides; Proteins; Protocols documentation; Reagent; Research; Resistance; Role; Science; Scientist; Serological; Site; Specificity; Staging; Structure; System; Testing; Titrations; Transcriptional Regulation; Vaccination; Vaccines; Variant; Vibrio cholerae; Work; base; experience; high risk; improved; in vivo; killings; novel; novel vaccines; null mutation; pathogen; prevent; public health relevance; sound; success

DESCRIPTION (provided by applicant): Vibrio cholerae (Vc) causes cholera. Its success as a pathogen depends on appropriate transcriptional regulation in different surroundings, such as the human intestinal tract or the aquatic environment, where growth conditions are sub-optimal. Gene regulation results in expression of operons that are critical for generation of a biofilm. Biofilms are complex structures composed of bacterial cells, proteins, and polysaccharides which allow the bacteria to exist in hostile environments. Compared to smooth Vc, the infectious rugose Vc variant is characterized by corrugated colonies, the capacity to develop extensive biofilms, and increased resistance to osmotic and oxidative stresses. Compared to smooth Vc, rugose Vc biofilms have enhanced survival in the presence of bile which may explain why recently shed Vc biofilm aggregates, that have biofilm characteristics, are more infectious for human. The rugosity and biofilm structure modulator gene cluster (rbmA-F) has been identified, which along with several other secreted proteins, are required for the formation of El Tor Vc rugose biofilms. The protective role of antibodies (Abs) to the Rbm proteins has not been studied. Rbm proteins are induced by bile and thus are expressed as part of the infecting inoculum and during the stages of colonization; RbmA, RbmC, and Bap-1 are required for wild type colonization. We hypothesize that the RbmA, RbmC, and Bap-1 (a homolog to RbmC) secreted proteins, known to regulate the biofilm matrix, are protective antigens able to induce antibodies (Abs) that modulate Vc biofilms and thus pathogenesis. These studies are important to determine if biofilm antigens are relevant targets for the immune system and thus cholera vaccines. The proposed study brings together the expertise of two scientists, Drs. Wade and Yildiz who both have extensive research experience in different aspects of cholera research. Dr. W. Wade is a molecular immunologist who pioneered the development of the vaccination protocols for synthetic Vc LPS antigens conjugated to carrier proteins. Dr. Wade is an established immunologist with an extensive track record in generating and characterizing protective Abs to Vc antigens (TcpA, LPS). Dr. F. Yildiz is a recognized molecular biologist who isolated and characterized the Rbm proteins (also referred to in the application as biofilm matrix or matrix proteins) and their requirement for Vc biofilm biology. PUBLIC HEALTH RELEVANCE: Cholera is still a disease that sickens millions and kills thousands yearly. We propose to formulate a new subunit cholera vaccine based on identification of protective epitopes in proteins known to be required for V. cholerae biofilm generation. Previous work from Dr. Wade's laboratories has shown that anti-TcpA antibodies can modify biofilm growth. Other work by Dr. Yildiz's group has identified proteins required for biofilm development that will serve as immunogens to induce antibodies.

描述（由申请人提供）：霍乱弧菌 (Vc) 引起霍乱。它作为病原体的成功取决于不同环境中适当的转录调节，例如人类肠道或水生环境，这些环境中的生长条件不是最理想的。基因调控导致操纵子的表达，这对于生物膜的生成至关重要。生物膜是由细菌细胞、蛋白质和多糖组成的复杂结构，使细菌能够在恶劣的环境中生存。与光滑 Vc 相比，传染性皱纹 Vc 变体的特点是具有波纹状集落、形成广泛生物膜的能力以及对渗透和氧化应激的抵抗力增强。与光滑的 Vc 相比，有皱纹的 Vc 生物膜在胆汁存在的情况下具有更高的存活率，这可以解释为什么最近脱落的具有生物膜特征的 Vc 生物膜聚集体对人类更具传染性。已鉴定出粗糙度和生物膜结构调节基因簇 (rbmA-F)，该基因簇与其他几种分泌蛋白一起是形成 El Tor Vc 粗糙度生物膜所必需的。抗体 (Abs) 对 Rbm 蛋白的保护作用尚未研究。 Rbm 蛋白由胆汁诱导，因此作为感染接种物的一部分并在定植阶段表达； RbmA、RbmC 和 Bap-1 是野生型定植所必需的。我们假设 RbmA、RbmC 和 Bap-1（RbmC 的同源物）分泌蛋白（已知可调节生物膜基质）是能够诱导抗体 (Ab) 的保护性抗原，从而调节 Vc 生物膜，从而调节发病机制。这些研究对于确定生物膜抗原是否是免疫系统以及霍乱疫苗的相关靶标非常重要。这项拟议的研究汇集了两位科学家的专业知识，博士。 Wade 和 Yildiz 在霍乱研究的不同方面都拥有丰富的研究经验。 W. Wade 博士是一位分子免疫学家，他率先开发了与载体蛋白缀合的合成 Vc LPS 抗原的疫苗接种方案。 Wade 博士是一位知名免疫学家，在生成和表征针对 Vc 抗原（TcpA、LPS）的保护性抗体方面拥有丰富的经验。 F. Yildiz 博士是一位公认的分子生物学家，他分离并表征了 Rbm 蛋白（在本申请中也称为生物膜基质或基质蛋白）及其对 Vc 生物膜生物学的要求。 公共卫生相关性：霍乱仍然是一种每年导致数百万人患病、数千人死亡的疾病。我们建议根据已知霍乱弧菌生物膜生成所需的蛋白质中的保护性表位的鉴定来配制一种新的亚单位霍乱疫苗。 Wade 博士实验室之前的工作表明，抗 TcpA 抗体可以改变生物膜的生长。 Yildiz 博士小组的其他工作已经确定了生物膜发育所需的蛋白质，这些蛋白质将作为免疫原来诱导抗体。