Production of a new tetravalent vaccine targeting fimbriae and enterotoxin of enterotoxigenic Escherichia coli.

ChongLi Xu, Yuhan She, Fengyang Fu, ChongBo Xu
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Abstract

Enterotoxigenic Escherichia coli (ETEC) is an important type of pathogenic bacteria that causes diarrhea in pigs. The objective of this study was to prepare a novel tetravalent vaccine to effectively prevent piglet diarrhea caused by E. coli. In order to realize the production of K88ac-K99-ST1-LTB tetravalent inactivated vaccine, the biological characteristics, stability, preservation conditions, and safety of the recombinant strain BL21(DE3) (pXKKSL4) were studied, and the vaccine efficacy and minimum immune dose were measured. The results indicated that the biological characteristics, target protein expression, and immunogenicity of the 1st to 10th generations of the strain were stable. Therefore, the basic seed generation was preliminarily set as the 1st to 10th generations. The results of the efficacy tests showed that the immune protection rate could reach 90% with 1 minimum lethal dose (MLD) virulent strain attack in mice. The immunogenicity was stable, and the minimum immune dose was 0.1 mL per mouse. Our research showed that the genetically engineered vaccine developed in this way could prevent piglet diarrhea caused by enterotoxigenic E. coli through adhesin and enterotoxin. In order to realize industrial production of the vaccine as soon as possible, we conducted immunological tests and production process research on the constructed K88ac-K99-ST1-LTB tetravalent inactivated vaccine. The results of this study provide scientific experimental data for the commercial production of vaccines and lay a solid foundation for their industrial production.

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针对肠毒性大肠杆菌的鞭毛和肠毒素生产新型四价疫苗。
肠毒性大肠杆菌(ETEC)是导致猪腹泻的一种重要致病菌。本研究旨在制备一种新型四价疫苗,以有效预防由大肠杆菌引起的仔猪腹泻。为实现K88ac-K99-ST1-LTB四价灭活疫苗的制备,研究了重组菌株BL21(DE3)(pXKKSL4)的生物学特性、稳定性、保存条件和安全性,并测定了疫苗效价和最小免疫剂量。结果表明,该菌株第1代至第10代的生物学特性、靶蛋白表达和免疫原性都很稳定。因此,初步确定基本种子代为第 1 至第 10 代。药效试验结果表明,以 1 个最小致死剂量(MLD)毒株攻击小鼠,免疫保护率可达 90%。免疫原性稳定,最小免疫剂量为每只小鼠 0.1 mL。我们的研究表明,用这种方法研制的基因工程疫苗可通过粘附素和肠毒素预防肠毒性大肠杆菌引起的仔猪腹泻。为了尽快实现疫苗的工业化生产,我们对构建的 K88ac-K99-ST1-LTB 四价灭活疫苗进行了免疫学试验和生产工艺研究。研究结果为疫苗的商业化生产提供了科学的实验数据,为疫苗的工业化生产奠定了坚实的基础。
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