Generation, safety and immunogenicity of an Actinobacillus pleuropneumoniae quintuple deletion mutant SLW07 (ΔapxICΔapxIICΔorf1ΔcpxARΔarcA)

Fangyan Yuan; Jinlin Liu; Wujin You; Weicheng Bei; Chunlai Wang; Jin Zhao; Yiongxiang Tian; Siguo Liu

doi:10.1016/j.vaccine.2018.02.083

Generation, safety and immunogenicity of an Actinobacillus pleuropneumoniae quintuple deletion mutant SLW07 (ΔapxICΔapxIICΔorf1ΔcpxARΔarcA)

Vaccine. 2018 Mar 27;36(14):1830-1836. doi: 10.1016/j.vaccine.2018.02.083. Epub 2018 Feb 24.

Authors

Fangyan Yuan¹, Jinlin Liu², Wujin You³, Weicheng Bei³, Chunlai Wang⁴, Jin Zhao², Yiongxiang Tian⁵, Siguo Liu⁶

Affiliations

¹ Institute of Animal Husbandry and Veterinary Sciences, Hubei Academy of Agricultural Sciences, Key Laboratory of Prevention and Control Agents for Animal Bacteriosis (Ministry of Agriculture), Hubei Key Laboratory of Animal Embryo and Molecular Breeding, Wuhan, Hubei 430064, China.
² College of Life Sciences, Central China Normal University, Wuhan, Hubei 430079, China.
³ State Key Laboratory of Agricultural Microbiology, The Cooperative Innovation Center for Sustainable Pig Production, College of Veterinary Medicine, Huazhong Agricultural University, Wuhan, Hubei 430070, China.
⁴ State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150001, China.
⁵ Institute of Animal Husbandry and Veterinary Sciences, Hubei Academy of Agricultural Sciences, Key Laboratory of Prevention and Control Agents for Animal Bacteriosis (Ministry of Agriculture), Hubei Key Laboratory of Animal Embryo and Molecular Breeding, Wuhan, Hubei 430064, China. Electronic address: tyxanbit@163.com.
⁶ State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150001, China. Electronic address: siguo_liu@hvri.ac.cn.

PMID: 29486946
DOI: 10.1016/j.vaccine.2018.02.083

Abstract

We inactivated a virulence determinant, ArcA, in an Actinobacillus pleuropneumoniae quadruple deletion mutant SLW06 (ΔapxICΔapxIICΔorf1ΔcpxAR, serovar 1), and a quintuple deletion mutant SLW07 was generated. SLW07 showed decreased adherence to and invasion of host cells, compared to its parent strain SLW06. SLW07 was more sensitive in RAW264.7 macrophage-mediated phagocytosis and clearance. SLW07 was less virulent in mice. An immunization assay indicated that both SLW07 and SLW06 preferentially stimulated T helper cell type 2 response in mice. Live vaccines induced the production of interleukin-6 and tumor necrosis factor-α by splenic lymphocytes. Furthermore, the protective immunity of SLW07 was not affected after ArcA mutation. Immunization with SLW07 could provide a complete protection following virulent A. pleuropneumoniae challenge in mice. Our results suggest that SLW07 is a promising live vaccine candidate, which is further attenuated from and shares similar protective efficacy with its quadruple deletion parent SLW06.

Keywords: Actinobacillus pleuropneumoniae; Protective efficacy; Quintuple deletion mutant; Safety.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Actinobacillus Infections / immunology*
Actinobacillus Infections / microbiology*
Actinobacillus Infections / prevention & control
Actinobacillus pleuropneumoniae / genetics*
Actinobacillus pleuropneumoniae / immunology*
Animals
Antibodies, Bacterial / immunology
Bacterial Adhesion / genetics
Bacterial Proteins / genetics*
Bacterial Proteins / immunology*
Bacterial Vaccines / genetics
Bacterial Vaccines / immunology
Cytokines / metabolism
Disease Models, Animal
Female
Gene Knockdown Techniques
Immunity, Humoral
Immunoglobulin G / immunology
Lung / immunology
Lung / microbiology
Lung / pathology
Macrophages / immunology
Macrophages / microbiology
Mice
Phagocytosis / immunology
Sequence Deletion*
Virulence / genetics
Virulence Factors

Substances

Antibodies, Bacterial
Bacterial Proteins
Bacterial Vaccines
Cytokines
Immunoglobulin G
Virulence Factors