Evaluation of the immunogenicity of auxotrophic Lactobacillus with CRISPR-Cas9D10A system-mediated chromosomal editing to express porcine rotavirus capsid protein VP4

Autor: Fengsai Li, Zhuyuan Mei, Ning Ju, Ling Sui, Xiaolong Fan, Zi Wang, Jiaxuan Li, Yanping Jiang, Wen Cui, Zhifu Shan, Han Zhou, Li Wang, Xinyuan Qiao, Lijie Tang, Xiaona Wang, Yijing Li
Jazyk: angličtina
Rok vydání: 2022
Předmět:
Zdroj: Virulence, Vol 13, Iss 1, Pp 1315-1330 (2022)
Druh dokumentu: article
ISSN: 21505594
2150-5608
2150-5594
DOI: 10.1080/21505594.2022.2107646
Popis: Porcine rotavirus (PoRV) is an important pathogen, leading to the occurrence of viral diarrhoea . As the infection displays obvious enterotropism, intestinal mucosal immunity is the significant line of defence against pathogen invasion. Moreover, as lactic acid bacteria (LAB) show acid resistance, bile salt resistance and immune regulation, it is of great significance to develop an oral vaccine. Most traditional plasmid delivery vectors use antibiotic genes as selective markers, easily leading to antibiotic accumulation. Therefore, to select a food-grade marker in genetically engineering food-grade microorganisms is vital. Based on the CRISPR-Cas9D10A system, we constructed a stable auxotrophic Lactobacillus paracasei HLJ-27 (Lactobacillus △Alr HLJ-27) strain. In addition, as many plasmids replicate in the host bacteria, resulting in internal gene deletions. In this study,we used a temperature-sensitive gene editing plasmidto insert the VP4 gene into the genome, yielding the insertion mutant strains VP4/△Alr HLJ-27, VP4/△Alr W56, and VP4/W56. This recombinant bacterium efficiently induced secretory immunoglobulin A (SIgA)-based mucosal and immunoglobulin G (IgG)-based humoral immune responses. These oral mucosal vaccines have the potential to act as an alternative to the application of antibiotics in the future and induce efficient immune responses against PEDV infection.
Databáze: Directory of Open Access Journals
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