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).
About author: TIAN Qiu-feng, E-mail: email@example.com
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.
Barrow P A, Freitas Neto O C. 2011. Pullorum disease and fowl typhoid - new thoughts on old diseases: A review. Avian Pathology, 40, 1-13.
Brogden K A. 2005. Antimicrobial peptides: Pore formers or metabolic inhibitors in bacteria? Nature Reviews Microbiology, 3, 238-250.
Chappell L, Kaiser P, Barrow P, Jones M A, Johnston C, Wigley P. 2009. The immunobiology of avian systemic salmonellosis. Veterinary Immunology and Immunopathology, 128, 53-59.
Chaudhari A A, Jawale C V, Kim S W, Lee J H. 2012. Construction of a Salmonella Gallinarum ghost as a novel inactivated vaccine candidate and its protective efficacy against fowl typhoid in chickens. Veterinary Research, 43, 44.
Chong C, Bost K L, Clements J D. 1996. Differential production of interleukin-12 mRNA by murine macrophages in response to viable or killed Salmonella spp. Infection and Immunity, 64, 1154-1160.
Flynn J L, Goldstein M M, Triebold K J, Koller B, Bloom B R. 1992. Major histocompatibility complex class I-restricted T cells are required for resistance to Mycobacterium tuberculosis infection. Proceedings of the National Academy of Sciences of the United States of America, 89, 12013-12017.
Gómez-Verduzco G, Tellez G, Quintana A L, Isibasi A, Ortiz-Navarrete V. 2010. Humoral immune response in breeding hens and protective immunity provided by administration of purified Salmonella Gallinarum porins. Poultry Science, 89, 495-500.
Haidinger W, Mayr U B, Szostak M P, Resch S, Lubitz W. 2003. Escherichia coli ghost production by expression of lysis gene E and staphylococcal nuclease. Applied and Environmental Microbiology, 69, 6106-6113.
Harty J T, Bevan M J. 1992. CD8+ T cells specific for a single nonamer epitope of Listeria monocytogenes are protective in vivo. Journal of Experimental Medicine, 175, 1531-1538.
Hensel A, Huter V, Katinger A, Raza P, Strnistschie C, Roesler U, Brand E, Lubitz W. 2000. Intramuscular immunization with genetically inactivated (ghosts) Actinobacillus pleuropneumoniae serotype 9 protects pigs against homologous aerosol challenge and prevents carrier state. Vaccine, 18, 2945-2955.
Jalava K, Hensel A, Szostak M P, Resch S, Lubitz W. 2002. Bacterial ghosts as vaccine candidates for veterinary applications. Journal of Controlled Release, 85, 17-25.
Jawale C V, Chaudhari A A, Lee J H. 2014. Generation of a safety enhanced Salmonella Gallinarum ghost using antibiotic resistance free plasmid and its potential as an effective inactivated vaccine candidate against fowl typhoid. Vaccine, 32, 1093-1099.
Jechlinger W, Glocker J, Haidinger W, Matis A, Szostak M P, Lubitz W. 2005. Modulation of gene expression by promoter mutants of the lambda cI857/pRM/pR system. Journal of Biotechnology, 116, 11-20.
Kwon S R, Kang Y J, Lee D J, Lee E H, Nam Y K, Kim S K, Kim K H. 2009. Generation of Vibrio anguillarum ghost by coexpression of PhiX 174 lysis E gene and staphylococcal nuclease A gene. Molecular Biotechnology, 42, 154-159.
Li Q, Hu Y, Chen J, Liu Z, Han J, Sun L, Jiao X. 2013. Identification of Salmonella enterica serovar Pullorum antigenic determinants expressed in vivo. Infection and Immunity, 81, 3119-3127.
Lo W F, Ong H, Metcalf E S, Soloski M J. 1999. T cell responses to Gram-negative intracellular bacterial pathogens: A role for CD8+ T cells in immunity to Salmonella infection and the involvement of MHC class Ib molecules. Journal of Immunology, 162, 5398-5406.
Lu S, Manges A R, Xu Y, Fang F C, Riley L W. 1999. Analysis of virulence of clinical isolates of Salmonella enteritidis in vivo and in vitro. Infection and Immunity, 67, 5651-5657.
Lundin B S, Johansson C, Svennerholm A M. 2002. Oral immunization with a Salmonella enterica serovar typhi vaccine induces specific circulating mucosa-homing CD4+ and CD8+ T cells in humans. Infection and Immunity, 70, 5622-5627.
Mader H J, Szostak M P, Hensel A, Lubitz W, Haslberger A G. 1997. Endotoxicity does not limit the use of bacterial ghosts as candidate vaccines. Vaccine, 15, 195-202.
Mayr U B, Haller C, Haidinger W, Atrasheuskaya A, Bukin E, Lubitz W, Ignatyev G. 2005. Bacterial ghosts as an oral vaccine: A single dose of Escherichia coli O157:H7 bacterial ghosts protects mice against lethal challenge. Infection and Immunity, 73, 4810-4817.
Mittrucker H W, Kaufmann S H. 2000. Immune response to infection with Salmonella typhimurium in mice. Journal of Leukocyte Biology, 67, 457-463.
Nguyen L T, Haney E F, Vogel H J. 2011. The expanding scope of antimicrobial peptide structures and their modes of action. Trends in Biotechnology, 29, 464-472.
Panthel K, Jechlinger W, Matis A, Rohde M, Szostak M P, Lubitz W, Haas R. 2003. Generation of Helicobacter pylori ghosts by PhiX protein E-mediated inactivation and their evaluation as vaccine candidates. Infection and Immunity, 71, 109-116.
Peng W, Si W, Yin L, Liu H, Yu S, Liu S, Wang C, Chang Y, Zhang Z, Hu S, Du Y. 2011. Salmonella enteritidis ghost vaccine induces effective protection against lethal challenge in specific-pathogen-free chicks. Immunobiology, 216, 558-565.
Salerno-Goncalves R, Pasetti M F, Sztein M B. 2002. Characterization of CD8+ effector T cell responses in volunteers immunized with Salmonella enterica serovar Typhi strain Ty21a typhoid vaccine. Journal of Immunology, 169, 2196-2203.
Sancho P, Tejedor C, Sidhu-Munoz R S, Fernandez-Lago L, Vizcaino N. 2014. Evaluation in mice of Brucella ovis attenuated mutants for use as live vaccines against B. ovis infection. Veterinary Research, 45, 61.
Shivaprasad H L. 2000. Fowl typhoid and pullorum disease. Revue Scientifique et Technique - Office International des Epizooties, 19, 405-424.
Si W, Yu S, Chen L, Wang X, Zhang W, Liu S, Li G. 2014. Passive protection against Salmonella enterica serovar Enteritidis infection from maternally derived antibodies of hens vaccinated with a ghost vaccine. Research in Veterinary Science, 97, 191-193.
Sood S, Rishi P, Dhawan V, Sharma S, Ganguly N K. 2005. Protection mediated by antibodies to iron-regulated outer-membrane proteins of S. typhi in a mouse peritonitis model. Molecular and Cellular Biochemistry, 273, 69-78.
Szostak M P, Hensel A, Eko F O, Klein R, Auer T, Mader H, Haslberger A, Bunka S, Wanner G, Lubitz W. 1996. Bacterial ghosts: Non-living candidate vaccines. Journal of Biotechnology, 44, 161-170.
Sztein M B, Tanner M K, Polotsky Y, Orenstein J M, Levine M M. 1995. Cytotoxic T lymphocytes after oral immunization with attenuated vaccine strains of Salmonella typhi in humans. Journal of Immunology, 155, 3987-3993.
Tabrizi C A, Walcher P, Mayr U B, Stiedl T, Binder M, McGrath J, Lubitz W. 2004. Bacterial ghosts--biological particles as delivery systems for antigens, nucleic acids and drugs. Current Opinion in Biotechnology, 15, 530-537.
White D W, Harty J T. 1998. Perforin-deficient CD8+ T cells provide immunity to Listeria monocytogenes by a mechanism that is independent of CD95 and IFN-γ but requires TNF-α. Journal of Immunology, 160, 898-905.
Wigley P, Berchieri A, Page K L, Smith A L, Barrow P A. 2001. Salmonella enterica serovar Pullorum persists in splenic macrophages and in the reproductive tract during persistent, disease-free carriage in chickens. Infection and Immunity, 69, 7873-7879.
Won G, Chaudhari A A, Lee J H. 2016. Protective efficacy and immune responses by homologous prime-booster immunizations of a novel inactivated Salmonella Gallinarum vaccine candidate. Clinical and Experimental Vaccine Research, 5, 148-158.
Yu S, Zhao H, Wang H, Wang X, Shao G, Xu L, Si W, Chen L, Zhang W, Liu S. 2013. Production and characterization of mouse monoclonal antibodies against lysis protein E of phiX174. Journal of Virological Methods, 189, 355-361.
Yu S Y, Peng W, Si W, Yin L, Liu S G, Liu H F, Zhao H L, Wang C L, Chang Y H, Lin Y Z. 2011. Enhancement of bacteriolysis of shuffled phage PhiX174 gene E. Virology Journal, 8, doi: 10.1186/1743-422X-8-206
Zhu J, Yamane H, Paul W E. 2010. Differentiation of effector CD4 T cell populations. Annual Review of Immunology, 28, 445-489.