Development of rotavirus-based enterotoxigenic Escherichia coli dual vaccines

基于轮状病毒的产肠毒素大肠杆菌双重疫苗的研制

• 批准号: 10741541
• 负责人: Siyuan Ding
• 金额: $ 27.21万
• 依托单位: WASHINGTON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-08-16 至 2025-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10741541
• 关键词: Acquired Immunodeficiency Syndrome; Acute; Adult; Affect; Age; Animal Model; Antibodies; Antibody Response; Antigens; Antiviral Agents; Astrovirus; Atrophic; Attenuated; Automobile Driving; Biological Assay; Biology; Body Weight decreased; Body Weights and Measures; Cessation of life; Child; Colony-Forming Units Assay; Country; Cryptosporidium; Cyclic AMP; Cyclic GMP; Data; Development; Diarrhea; Disease; Double-Stranded RNA; Enteral; Enterotoxins; Enzyme-Linked Immunosorbent Assay; Equilibrium; Escherichia coli Infections; Etiology; Future; Gastroenteritis; Genetic Engineering; Genome; Goals; Growth; Harvest; Heterophile Antigens; Human; Immune response; Immunity; Immunize; Immunofluorescence Immunologic; Immunoglobulin A; Immunoglobulin G; Infant; Infection; Intestines; Laboratories; Licensing; Malaria; Measles; Measures; Medical; Methods; Monitor; Morbidity - disease rate; Mucous Membrane; Mus; Neonatal; Norovirus; Oral; Organoids; Outcome; Pathogenesis; Plaque Assay; RNA vaccine; Recombinants; Reporter; Resolution; Rotavirus; Rotavirus Infections; Rotavirus VP6 protein; Rotavirus Vaccines; Series; Serum; Shigella; Small Intestines; Specimen; System; Testing; Therapeutic Intervention; Tissues; Toxic effect; Toxin; Toxoids; Vaccinated; Vaccination; Vaccines; Vertebral column; Villus; Viral; Viral Vector; Virulence; Virulent; Virus; Virus Replication; Viviparous-1 protein; co-infection; cognitive development; design; diarrheal disease; efficacy evaluation; enteric infection; enteric pathogen; enterotoxigenic Escherichia coli; immunogenic; immunogenicity; improved; innovation; mortality; mouse model; mucosal vaccine; mutant; nanoluciferase; neonatal mice; neutralizing antibody; novel; pathogen; prevent; protective efficacy; pup; rational design; response; reverse genetics; suckling; tissue culture; vaccine candidate; vaccine efficacy; vaccine platform; vector; vector induced; vector vaccine

Project Summary Diarrheal diseases account for 1 in 10 child deaths worldwide. Rotavirus (RV) and enterotoxigenic Escherichia coli (ETEC) are major etiological causes of acute gastroenteritis and severe diarrhea worldwide, together resulting in approximately 300,000 deaths each year, mostly in children under the age of five. Current RV vaccines have limited efficacy in endemic countries and no direct antivirals are available. No vaccine is licensed for ETEC. Our overall objectives are to better understand the biology of RV and ETEC and to use that information to develop therapeutic interventions to alleviate diarrhea and sequelae. In preliminary studies, we utilized an improved RV reverse genetics system developed by our lab and generated a recombinant murine RV that encodes the protruding domain of the human norovirus VP1 protein. We found that this RV-based viral vector induces a robust antigen-specific serum IgG and fecal IgA responses in inoculated mouse pups. Based on these data, we constructed several new recombinant RVs that express ETEC heat-labile and heat-stable toxins. We hypothesize that one or more novel RV-ETEC dual vaccine candidates will induce a protective humoral immune response in mice and reduce pathogen burden and pathogenesis from subsequent RV and ETEC infections. To test this hypothesis, we have developed a highly tractable murine RV reverse genetics method, disease relevant neonatal and adult mouse models, and several innovative primary human small bowel organoid cultures, which will provide an unprecedented resolution of understanding of the immunogenicity and protective efficacy of our vaccine candidates. In Aim 1, we will characterize RV replication and define the immunological responses of our dual vaccines in neonatal mice. In Aim 2, we will examine the efficacy of the recombinant RV- ETEC dual vaccines in protecting immunized mice from RV and ETEC challenges in adult mice. Collectively, we expect our proof-of-principle study to start to establish the utility of RVs as an innovative live-attenuated mucosal vaccine platform to encode foreign antigens and broadly protect against common enteric pathogens.

项目摘要 腹泻疾病在全球十分之一的儿童死亡中占1个。轮状病毒（RV）和肠毒素 大肠杆菌（ETEC）是全球急性胃炎和严重腹泻的主要病因原因， 每年共同导致大约300,000人死亡，主要是五岁以下的儿童。当前的 RV疫苗在地方性国家的疗效有限，没有直接抗病毒药。没有疫苗 获得ETEC的许可。我们的总体目标是更好地了解RV和ETEC的生物学，并使用它 信息以开发治疗干预措施以减轻腹泻和后遗症。 在初步研究中，我们利用了我们实验室开发的改进的RV反向遗传系统， 产生了一个重组鼠RV，该鼠RV编码人诺如病毒VP1蛋白的突出结构域。 我们发现，基于RV的病毒载体会诱导强大的抗原特异性血清IgG和粪便IgA反应 在接种的小鼠幼崽中。基于这些数据，我们构建了几种表达的新重组RV ETEC热能和热稳定毒素。我们假设一种或多种新型RV-ETEC双疫苗 候选者将诱导小鼠保护性体液免疫反应并减轻病原体负担 以及随后的RV和ETEC感染的发病机理。 为了检验这一假设，我们已经开发了一种高度可触犯的鼠RV反向遗传学方法，疾病 相关的新生儿和成人小鼠模型，以及几种创新的原代人小肠类正弦 培养物将提供对免疫原性和保护性理解的前所未有的解决方案 我们候选疫苗的功效。在AIM 1中，我们将表征RV复制并定义免疫学 我们在新生小​​鼠中双重疫苗的反应。在AIM 2中，我们将检查重组RV-的功效 ETEC双重疫苗保护免疫小鼠免受成年小鼠的RV和ETEC挑战。共同 我们希望我们的原理学研究能够开始建立RVS的实用性 粘膜疫苗平台编码外国抗原并广泛预防常见的肠道病原体。