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 腹泻病占全世界儿童死亡人数的十分之一。轮状病毒（RV）和肠毒素 大肠杆菌（ETEC）是全球急性胃肠炎和严重腹泻的主要病原学原因， 每年造成大约30万人死亡，其中大多数是5岁以下的儿童。电流 RV疫苗在流行国家的效力有限，并且没有直接的抗病毒药物。没有一种疫苗是 获得ETEC许可。我们的总体目标是更好地了解RV和ETEC的生物学， 提供信息，以制定治疗干预措施，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， 产生编码人诺如病毒VP 1蛋白的突出结构域的重组鼠RV。 我们发现，这种基于RV的病毒载体诱导了强有力的抗原特异性血清IgG和粪便伊加应答 在接种的小鼠幼崽中。基于这些数据，我们构建了几个新的重组RV， ETEC热不稳定和热稳定毒素。我们假设一种或多种新型RV-ETEC双重疫苗 候选物将在小鼠中诱导保护性体液免疫应答， 以及随后RV和ETEC感染的发病机制。 为了验证这一假设，我们已经开发了一种高度易处理的小鼠RV反向遗传学方法， 相关的新生儿和成年小鼠模型，以及几种创新的原代人小肠类器官 这将为理解免疫原性和保护性提供前所未有的解决方案。 我们候选疫苗的有效性。在目标1中，我们将描述RV复制的特征，并定义免疫学上的 我们的双重疫苗在新生小鼠中的反应。在目标2中，我们将检查重组RV的功效- ETEC双重疫苗在成年小鼠中保护免疫小鼠免受RV和ETEC攻击。总的来说， 我们希望我们的原理验证研究能够开始建立RV作为一种创新的减毒活疫苗的实用性， 粘膜疫苗平台编码外源抗原并广泛保护抵抗常见肠道病原体。