Construction of Salmonella Pullorum ghost by co-expression of lysis gene E and the antimicrobial peptide SMAP29 and evaluation of its immune efficacy in specific-pathogen-free chicks
TIAN Qiu-feng1, 2*, ZHOU Wei1, 2*, SI Wei1, YI Fei1, 2, HUA Xin1, YUE Min3, CHEN Li-ping1, LIU Si-guo1, YU Shen-ye1
1 Division of Bacterial Diseases, State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150069, P.R.China 2 Heilongjiang Bayi Agricultural University, Daqing 163319, P.R.China 3 College of Animal Sciences, Zhejiang University, Hangzhou 310058, P.R.China
Abstract In this study, a safety enhanced Salmonella Pullorum (S. Pullorum) ghost was constructed using an antimicrobial peptide gene, and evaluated for its potential as a Pullorum disease (PD) vaccine candidate. The antimicrobial peptide SMAP29 was co-expressed with lysis gene E to generate S. Pullorum ghosts. No viable bacteria were detectable either in the fermentation culture after induction of gene E- and SMAP29-mediated lysis for 24 h or in the lyophilized ghost products. Specific-pathogen-free (SPF) chicks were intraperitoneally immunized with ghosts at day 7 of age and no mortality, clinical symptoms or signs of PD such as anorexia, depression and diarrhea were observed. On challenge with a virulent S. Pullorum strain at 4 wk post-immunization, a comparatively higher level of protection was observed in the S. Pullorum ghost immunized chickens with a minimum of pathological lesions and bacterial loads compared to the birds in inactivated vaccine groups. In addition, immunization with the S. Pullorum ghosts induced a potent systemic IgG response and was associated with significantly increased levels of cytokine IFN-γ and IL-4 and relative percentages of CD4+ and CD8+ T lymphocytes. Our results indicate that SMAP29 can be employed as a new secondary lethal protein to enhance the safety of bacterial ghosts, and to prepare a non-living bacterial vaccine candidate that can prevent PD in chickens.
This work was supported by grants from the National Key Research and Development Program of China (2016YFD0501608), the National Natural Science Foundation of China (31470893), the Special Fund for Agro-scientific Research in the Public Interest, China (201403054) and the National High Technology Research and Development Program of China (2011AA10A210).
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Cite this article:
TIAN Qiu-feng, ZHOU Wei, SI Wei, YI Fei, HUA Xin, YUE Min, CHEN Li-ping, LIU Si-guo, YU Shen-ye. Construction of Salmonella Pullorum ghost by co-expression of lysis gene E and the antimicrobial peptide SMAP29 and evaluation of its immune efficacy in specific-pathogen-free chicks[J]. Journal of Integrative Agriculture,
2018, 17(01): 197-209.
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