Determining vaccine candidacy of LT and STa toxoid fusions against enterotoxigeni

确定针对肠毒素的 LT 和 STa 类毒素融合疫苗的候选资格

• 批准号: 7860302
• 负责人: WEIPING ZHANG
• 金额: $ 20.66万
• 依托单位: SOUTH DAKOTA STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-06-05 至 2012-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7860302
• 关键词: Adult; Age; Animal Model; Animals; Antibodies; Antigens; Antitoxins; Attenuated; Attenuated Live Virus Vaccine; Bacterial Adhesins; Biological; Biotechnology; Cells; Cessation of life; Child; Clinical; Developing Countries; Development; Diarrhea; Disease; Economics; Enterotoxins; Epidemiology; Epithelium; Escherichia coli; Escherichia coli Infections; Escherichia coli Vaccines; Family suidae; Genes; Genetic; Gnotobiotic; Goals; Health; Heating; Human; Immune; Immunity; Immunocompromised Host; Immunoglobulin A; Immunoglobulin G; Individual; Infection; Ingestion; International; Intestines; Lead; Length; Life; Liquid substance; Mediating; Military Personnel; Modeling; Molecular; Molecular Biology; Mucosal Immunity; Mutate; National Institute of Allergy and Infectious Disease; Neutralization Tests; Oryctolagus cuniculus; Outcome; Outcome Study; Patients; Peptides; Population; Prevention; Prevention Research; Prevention strategy; Production; Research; Serum; Small Intestines; Study models; Subunit Vaccines; System; Time; Toxic effect; Toxin; Toxoids; Vaccines; Virulence; Virulence Factors; Virulent; antigen challenge; colonization factor antigens; disorder control; enterotoxigenic Escherichia coli; fimbria; immunogenic; immunogenicity; mutant; pathogen; prevent; programs; public health relevance; receptor; success; treatment strategy; vaccine development

DESCRIPTION (provided by applicant): Enterotoxigenic Escherichia coli (ETEC) strains are categorized as priority B pathogens by NIAID. ETEC-associated diarrhea is responsible for nearly one million deaths each year around the world in addition to causing of illness to immunocompromised individuals and travelers including deployed military personnel. The key virulence factors in ETEC- associated diarrhea are bacterial adhesins or colonization factor antigens (CFAs) and enterotoxins. CFAs mediate bacterial attachment to the host epithelium cells, but it is the enterotoxin, the heat-labile (LT) and heat-stable (STa), that stimulate fluid hyper-secretion and cause diarrhea. Early studies using LT and STa antigens for developing antitoxin vaccines had limited success because of the high toxicity of both toxins, non-immunogenicity of STa antigen, and lack of a suitable animal study model. Current advance in molecular biology makes it feasible to eliminate or reduce toxicity and to enhance STa immunogenicity. In this study, we will genetically mutate both LT and STa genes for expressing non- or low toxic LT and STa toxoids, and then genetically fuse these two mutated genes for LT and STa toxoid fusion antigens to enhance STa immunogenicity. These fusion antigens are expected to induce anti-LT and anti-STa antibodies, and those antibodies will neutralize native LT and STa toxins thus provide protection against ETEC infection. In our preliminary studies, we demonstrated that a genetic fusion of a porcine LT toxoid with a STa toxoid or STb (another non-immunogenic heat stable toxin) induced anti-LT and anti-ST antibodies, and these antibodies are protective against ETEC infection in a piglet challenge model. Piglet model, especially the system using piglets expressing K88ac receptors and ETEC strains expressing K88ac fimbriae, is perhaps the best model to study human ETEC associated diarrhea. K88ac receptor positive piglets are neutrally susceptible to ETEC strains expressing K88ac fimbriae, and develop identical clinical disease after infection of ETEC strains as human diarrheal patients. Furthermore, using pig model allows us to examine intestinal immunity, which is critical for providing protection against this disease. Our expertise in applying pig model in studying ETEC makes our research team the best candidate to conduct ETEC diarrhea research and prevention using the pig model. Determination vaccine candidacy of LT and STa toxoid fusions against ETEC diarrhea will provide essential information for antitoxin vaccine development, and will be very instructive in developing prevention and treatment strategies to protect humans and animals from diarrhea disease. PUBLIC HEALTH RELEVANCE: Enterotoxigenic Escherichia coli (ETEC) are the primary cause of diarrheal disease. ETEC diarrhea is responsible for near one million deaths each year, and brings major health problems and economic consequences in immunocompromised individuals (especially children under age of five years), international travelers and military personnel. Heat labile (LT) and heat-stable (STa) enterotoxins produced by ETEC strains are the virulence determinants in diarrheal infection. LT and STa stimulate fluid hyper-secretion in guts that results in diarrhea. Yet, there are no vaccines available to control or prevent this disease. Development of antitoxin vaccines could be an effective strategy to prevent diarrhea. However, both LT and STa are sufficiently toxic, thereby they cannot be used directly as antigens for developing antitoxin vaccines. This proposed research is to construct LT and STa toxoids, and to determine vaccine candidacy of fusion antigens from a full-length LT toxoid and a full-length STa toxoid against ETEC infection in a piglet model. The determination of vaccine candidacy of LT and STa toxoid fusions against diarrheal disease will provide essential information for developing effective antitoxin vaccines to prevent or control this disease.

性状（由申请方提供）：NIAID将产肠毒素大肠杆菌（ETEC）菌株归类为优先B类病原体。ETEC相关的腹泻每年在世界各地造成近100万人死亡，此外还导致免疫功能低下的个人和旅行者（包括部署的军事人员）患病。ETEC相关性腹泻的关键毒力因子是细菌粘附素或定植因子抗原（CFA）和肠毒素。CFA介导细菌附着到宿主上皮细胞，但肠毒素、热不稳定（LT）和热稳定（STa）刺激液体分泌过多并引起腹泻。早期使用LT和STa抗原开发抗毒素疫苗的研究成功有限，因为两种毒素的高毒性、STa抗原的非免疫原性以及缺乏合适的动物研究模型。目前分子生物学的进展使得消除或降低STa的毒性和增强其免疫原性成为可能。在本研究中，我们将对LT和STa基因进行遗传突变以表达无毒或低毒的LT和STa类毒素，然后将这两个突变的基因融合为LT和STa类毒素融合抗原以增强STa的免疫原性。预期这些融合抗原诱导抗LT和抗STa抗体，并且这些抗体将中和天然LT和STa毒素，从而提供针对ETEC感染的保护。在我们的初步研究中，我们证明了猪LT类毒素与STa类毒素或STb（另一种非免疫原性热稳定毒素）的基因融合诱导了抗LT和抗ST抗体，并且这些抗体在仔猪攻击模型中对ETEC感染具有保护作用。猪模型，尤其是利用表达K88ac受体的猪和表达K88ac菌毛的ETEC菌株的系统，可能是研究人类ETEC相关性腹泻的最佳模型。K88ac受体阳性仔猪对表达K88ac菌毛的ETEC菌株呈中性易感，感染ETEC菌株后发生与人类腹泻患者相同的临床疾病。此外，使用猪模型使我们能够检查肠道免疫，这对于提供针对这种疾病的保护至关重要。我们在应用猪模型研究ETEC方面的专业知识使我们的研究团队成为使用猪模型进行ETEC腹泻研究和预防的最佳候选人。确定LT和STa类毒素融合疫苗的候选人资格将为抗毒素疫苗的开发提供必要的信息，并将对制定预防和治疗策略以保护人类和动物免受腹泻疾病的影响具有重要的指导意义。公共卫生相关性：产肠毒素大肠杆菌（ETEC）是大肠杆菌病的主要原因。ETEC腹泻每年造成近100万人死亡，并给免疫功能低下的个人（特别是5岁以下的儿童）、国际旅行者和军事人员带来重大的健康问题和经济后果。ETEC菌株产生的热不稳定（LT）和热稳定（STa）肠毒素是大肠杆菌感染的毒力决定因素。LT和STa刺激肠道中的液体分泌过多，导致腹泻。然而，目前还没有疫苗可以控制或预防这种疾病。开发抗毒素疫苗可能是预防腹泻的有效策略。然而，LT和STa都具有足够的毒性，因此它们不能直接用作开发抗毒素疫苗的抗原。本研究拟构建LT类毒素和STa类毒素，并在仔猪模型中确定来自全长LT类毒素和全长STa类毒素的融合抗原的疫苗候选性。LT和STa类毒素融合疫苗候选人的确定将为研制有效的抗毒素疫苗预防或控制该病提供必要的信息。

项目成果

Modified heat-stable toxins (hSTa) of enterotoxigenic Escherichia coli lose toxicity but display antigenicity after being genetically fused to heat-labile toxoid LT(R192G).

• DOI: 10.3390/toxins3091146
• 发表时间: 2011-09
• 期刊: Toxins
• 影响因子: 4.2
• 作者: Liu M;Zhang C;Mateo K;Nataro JP;Robertson DC;Zhang W
• 通讯作者: Zhang W

Toxicity and immunogenicity of Enterotoxigenic Escherichia coli heat-labile and heat-stable toxoid fusion 3xSTa(A14Q)-LT(S63K/R192G/L211A) in a murine model.

• DOI: 10.1371/journal.pone.0077386
• 发表时间: 2013
• 期刊: PloS one
• 影响因子: 3.7
• 作者: Zhang C;Knudsen DE;Liu M;Robertson DC;Zhang W;STa Toxoid Vaccine Consortium Group
• 通讯作者: STa Toxoid Vaccine Consortium Group