Oral immunity and adjuvant receptors

口腔免疫和佐剂受体

• 批准号: 8288025
• 负责人: Georgios Hajishengallis
• 金额: $ 38.78万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-02-01 至 2015-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8288025
• 关键词: Address; Adjuvant; Adjuvanticity; Antibody Formation; Area; Binding; Biological; Bone Marrow; Buffaloes; CD4 Positive T Lymphocytes; Collaborations; Communicable Diseases; Complex; Crystallography; Data; Dendritic Cells; Dependence; Disease; Drug Formulations; Engineering; Enterotoxins; Escherichia coli; Exhibits; Funding; Gangliosides; Genitourinary system; Goals; Health; Heating; Immunity; Immunization; In Vitro; Infection; Inflammatory; Institutes; Invaded; Ligands; Lipid Binding; Masks; Mediating; Membrane Microdomains; Modeling; Molecular; Molecular Biology; Mucosal Immunity; Mus; Mutation; Oral; Paper; Parents; Periodontitis; Porphyromonas gingivalis; Pre-Clinical Model; Property; Proteins; Qualifying; Relative (related person); Research; Research Personnel; Roentgen Rays; Serum; Signal Transduction; Site; Solutions; Solvents; Structure; Surface; TLR1 gene; Technology; Testing; Time; Toll-Like Receptor 2; Toll-like receptors; Toxic effect; Universities; Vaccination; Vaccine Adjuvant; Vaccines; Work; base; improved; in vivo; innovation; interdisciplinary collaboration; mouse model; mutant; novel; novel vaccines; oral immunity; oral infection; pathogen; pre-clinical; public health relevance; receptor; respiratory; structural biology

DESCRIPTION (provided by applicant): This application addresses the compelling need to develop new and improved vaccine adjuvants for stimulating protective immunity at mucosal surfaces. The proposal involves a consortium arrangement between the University of Louisville, the University at Buffalo, and the Hauptman-Woodward Institute, and brings together complementary and integrated expertise in vaccine adjuvant research, structural and molecular biology, and induction of mucosal immunity. Studies by this group have succeeded to dissociate useful adjuvant properties from the toxicity of LT-IIb, an AB5-type heat-labile enterotoxin of Escherichia coli. Specifically, the recombinantly expressed B pentameric subunit of LT-IIb (LT-IIb-B5) not only lacks enterotoxicity but, strikingly, can activate the Toll-like receptor (TLR)-2/TLR1 signaling complex and stimulate antibody responses to co-administered vaccine proteins. Proof-of-concept preliminary studies have shown that certain engineered point-substitution mutations in the active region of LT-IIb-B5 enhance its ability to interact with TLR2 and TLR1. The overall objective of this application is to better characterize the physical and functional interaction of LT-IIb-B5 with the TLR2/TLR1 heterodimer, engineer improved versions of LT-IIb-B5 in terms of TLR2/TLR1-mediated adjuvanticity, and develop potential adjuvants through studies in preclinical models of vaccination against oral infection. This will be accomplished through appropriate and innovative molecular biological, structural, and immunological studies outlined in the following four specific aims: 1) To rationally engineer enhanced TLR2-interactive versions of LT-IIb-B5. 2) To structurally characterize the LT-IIb- B5-TLR2/TLR1 interaction using small-angle X-ray scattering (SAXS). 3) To evaluate the engineered LT-IIb-B5 mutants for in vitro TLR2/TLR1-dependent immunostimulatory activities. 4) To determine the ability of LT-IIb-B5 and improved engineered versions thereof to induce protective immunity in vivo. The generated data will form the basis for the long-term goal which is to establish LT-IIb-B5 and improved engineered derivatives as effective adjuvants in vaccine formulations against pathogens which colonize or invade via oral, respiratory, or urogenital mucosal surfaces. PUBLIC HEALTH RELEVANCE: The objective of this application is to rationally engineer novel Toll-like receptor-dependent mucosal adjuvants. These constructs will be evaluated for their capacity to stimulate protective immunity using appropriate preclinical mouse models. The focus of this proposal is on a novel vaccine against periodontitis, an oral inflammatory disease with an impact on systemic health, although the developed mucosal adjuvants can also be used in vaccine formulations against pathogens which colonize or invade via oral, respiratory, or urogenital mucosal surfaces. This work is timely and important given the paucity of safe and effective vaccine adjuvants and the tremendous health impact of infectious diseases.

描述（由申请人提供）：此申请解决了开发新的和改进的疫苗佐剂的迫切需求，以刺激粘膜表面的保护性免疫。该提案涉及路易斯维尔大学，布法罗大学和Hauptman-Woodward Institute之间的联盟安排，并汇集了疫苗辅助研究，结构和分子生物学的补充和综合专业知识，以及诱导粘膜免疫。该组的研究成功地将有用的辅助性质与LT-IIB的毒性分离出来，LT-IIB是Escherichia coli的AB5型热labile肠毒素。具体而言，LT-IIB（LT-IIB-B5）重组表达的B五聚体亚基不仅缺乏肠毒性，而且令人惊讶的是，可以激活类似Toll的受体（TLR）-2/TLR1信号传导复合物，并刺激对共同加速疫苗蛋白蛋白的抗体反应。概念验证的初步研究表明，LT-IIB-B5活性区域中某些工程的点固定突变增强了其与TLR2和TLR1相互作用的能力。该应用程序的总体目的是更好地表征LT-IIB-B5与TLR2/TLR1异二聚体的物理和功能相互作用，在TLR2/TLR2/TLR1介导的辅助性方面，工程师改进了LT-IIB-B5的版本，并通过预先策略模型在预先的抗性抗抗抗抗癌抗抗中，以开发潜在的辅助感。这将通过以下四个特定目标中概述的适当和创新的分子生物学，结构和免疫学研究来实现：1）合理工程的LT-IIB-B5的TLR2相互作用版本增强。 2）使用小角度X射线散射（SAXS）在结构上表征LT-IIB-B5-TLR2/TLR1相互作用。 3）评估工程的LT-IIB-B5突变体的体外TLR2/TLR1依赖性免疫刺激活性。 4）确定LT-IIB-B5和改进其工程版本在体内诱导免疫力的能力。生成的数据将构成长期目标的基础，该目标是建立LT-IIB-B5并改善工程衍生物作为疫苗配方中的有效佐剂，以针对通过口服，呼吸道或泌尿生殖器粘膜表面殖民或入侵的病原体。 公共卫生相关性：该应用的目的是合理地工程师新颖的Toll-Toll样受体依赖性粘膜佐剂。这些结构将通过适当的临床前小鼠模型来评估它们刺激保护性免疫的能力。该提案的重点是针对牙周炎的新型疫苗，这是一种对系统健康影响的口腔炎性疾病，尽管发达的粘膜佐剂也可以用于疫苗配方中，以针对病原体，该病原体通过口服，呼吸道或泌尿原质粘膜表面定植或入侵。考虑到安全有效的疫苗佐剂以及传染病的巨大健康影响，这项工作是及时且重要的。