JMB :: Journal of Microbiology and Biotechnology

Author	Renee Cornford-Nairns, Grant Daggard, Trilochan Mukkur
Affiliation	Department of Biological and Physical Sciences, University of Southern Queensland, Toowoomba, 4350 Queensland, Australia
Title	Construction and Preliminary Immunobiological Characterization of a Novel, Non-Reverting, Intranasal Live Attenuated Whooping Cough Vaccine Candidate
PublicationInfo	J. Microbiol. Biotechnol.2012 ; 22(6): 856~865
Abstract	We describe the construction and immunobiological properties of a novel whooping cough vaccine candidate, in which the aroQ gene, encoding 3-dehydroquinase, was deleted by insertional inactivation using the kanamycin resistance gene cassette and allelic exchange using a Bordetella suicide vector. The aroQ B. pertussis mutant required supplementation of media to grow but failed to grow on an unsupplemented medium. The aroQ B. pertussis mutant was undetectable in the trachea and lungs of mice at days 6 and 12 post-infection, respectively. Antigen-specific antibody isotypes IgG1 and IgG2a, were produced, and cell-mediated immunity [CMI], using interleukin-2 and interferon-gamma as indirect indicators, was induced in mice vaccinated with the aroQ B. pertussis vaccine candidate, which were substantially enhanced upon second exposure to virulent B. pertussis. Interleukin- 12 was also produced in the aroQ B. pertussis-vaccinated mice. On the other hand, neither IgG2a nor CMI-indicator cytokines were produced in DTaP-vaccinated mice, although the CMI-indicator cytokines became detectable post-challenge with virulent B. pertussis. Intranasal immunization with one dose of the aroQ B. pertussis mutant protected vaccinated mice against an intranasal challenge infection, with no pathogen being detected in the lungs of immunized mice by day 7 post-challenge. B. pertussis aroQ thus constitutes a safe, non-reverting, metabolite-deficient vaccine candidate that induces both humoral and cellmediated immune responses with potential for use as a single-dose vaccine in adolescents and adults, in the first instance, with a view to disrupting the transmission cycle of whooping cough to infants and the community.
Full-Text(PDF)
Keywords	Bordetella pertussis, aroQ, live attenuated pertussis vaccine, cell-mediated immunity induction, antibody response, protection against pertussis

References

Andrews, R., A. Herceg, and C. Roberts. 1997. Pertussis notifications in Australia, 1991 to 1997. Commun. Dis. Intell. 21: 145-148.
Canthaboo, C., L. Williams, D. K. Xing, and M. J. Corbel. 2000. Investigation of cellular and humoral immune responses to whole cell and acellular pertussis vaccines. Vaccine 19: 637-643.
Cohen, S. M. and M. W. Wheeler. 1946. Pertussis vaccine prepared with phase-I cultures grown in fluid medium. Am. J. Pub. Health 36: 371-376.
Conway, M. A., L. Madrigal-Estebas, S. McClean, D. J. Brayden, and K. H. Mills. 2001. Protection against Bordetella pertussis infection following parenteral or oral immunisation with antigens entrapped in biodegradable particles: Effect of formulation and route of immunization on induction of Th1 and Th2 cells. Vaccine 19: 1940-1950.
Cornford, R. A. 1998. Characterisation of the aroD and/or aroQ gene of Bordetella pertussis. BSc. Hons. Thesis, University of Southern Queensland, Toowoomba, Queensland, Australia.
Cornford, R. A. 2003. A novel, genetically engineered intranasal whooping cough vaccine. PhD Thesis, University of Southern Queensland, Toowoomba, Queensland, Australia.
de Melker, H. E., M. A. Conyn-van Spaendonck, H. C. Rumke, J. K. van Wijngaarden, F. R. Mooi, and J. F. P. Schellekens. 1997. Pertussis in The Netherlands: An outbreak despite high levels of immunization with whole-cell vaccine. Emerg. Infect. Dis. 3:175-178.
Elahi, S., J. Holmstro, and V. Gerdts. 2007. The benefits of using diverse animal models for studying pertussis. Trends Microbiol. 15: 462-468.
Ermens, A. A. M., A. J. M. Bayens, A. C. M. van Gemert, and J. L. P. van Duijnhoven. 1997. Simple dot-blot method evaluated for detection of antibodies against extractable nuclear antigens. Clin. Chem. 43: 2420-2422.
Frohlich, B. T., E. R. De Bernardez Clark, G. R. Siber, and R. W. Swartz. 1995. Improved pertussis toxin production by Bordetella pertussis through adjusting the growth medium’s ionic composition. J. Biotech. 39: 205-219.
Fry, S. R., A. Y. Chen, G. Daggard, and T. K. Mukkur. 2008. Parenteral immunization of mice with a genetically inactivated pertussis toxin DNA vaccine induces cell-mediated immunity and protection. J. Med. Microbiol. 57: 28-35.
Gold, M. S., S. Noonan, M. Osbourn, S. Precepa, and A. E. Kempe. 2003. Local reactions at the fourth dose of acellular pertussis vaccine in South Australia. Med. J. Aust. 179: 191-194.
Hoiseth, S. K. and B. A. D. Stocker. 1981. Aromatic-dependent Salmonella Typhimurium are non-virulent and effective as live vaccines. Nature 291: 238-239.
He, Q. and J. Mertsola. 2008. Factors contributing to pertussis resurgence. Future Microbiol. 3: 329-339.
Kolos, V., R. Menzies, and P. McIntyre. 2007. Higher pertussis hospitalization rates in indigenous Australian infants, and delayed vaccination. Vaccine 25: 588-590.
Lalonde, G., P. D. O’Hanley, B. A. D. Stocker, and K. T. Denich. 1994. Characterisation of a 3-dehydroquinase gene from Actinobacillus pleuropneumoniae with homology to the eukaryotic genes qa-2 and QUTE. Mol. Microbiol. 11: 273-280.
Leef, M., K. L. Elkin, J. Barbic, and R. D. Shahin. 2000. Protective immunity to Bordetella pertussis requires both B cells and CD4+ T cells for key functions other than specific antibody production. J. Exp. Med. 191: 1841-1852.
Locht, C., R. Antoine, D. Raze, N. Mielcarek, D. Hot, Y. Lemoine, and Mascart F. 2004. Bordetella pertussis: From functional genomics to intranasal vaccination. Int. J. Med. Microbiol. 293: 583-588.
Mahon, B. P., M. T. Brady, and K. H. Mills. 2000. Protection against Bordetella pertussis in mice in the absence of detectable circulating antibody: Implications for long-term immunity in children. J. Infect. Dis. 181: 2087-2091.
Marshall, H. S., M. S. Gold, R. Gent, P. J. Quinn, L. Piotto, M. F. Clarke, and D. M. Roberton. 2006. Ultrasound examination of extensive limb swelling reactions after diphtheria-tetanusacellular pertussis or reduced-antigen content diphtheria-tetanusacellular pertussis immunization in preschool-aged children. Pediatrics 118: 1501-1509.
Marzouqi, I., P. Richmond, S. Fry, J. Wetherall, and T. Mukkur. 2010. Development of improved vaccines against whooping cough: Current status. Hum. Vaccin. 6: 1-11.
Mielcarek, N., G. Riveau, F. Remoue, R. Antoine, A. Capron, and C. Locht. 1998. Homologous and heterologous protection after single intranasal administration of live attenuated recombinant Bordetella pertussis. Nat. Biotechnol. 16: 454-457.
Mielcarek, N., A.-S. Debrie, D. Raze, J. Bertout, C. Rouanet, A. B. Younes, et al. 2006. Live attenuated B. pertussis as a singledose nasal vaccine against whooping cough. PLOS Pathog. 2:e65.
Mills, K. H. G. 2001. Immunity to Bordetella pertussis. Microbes Infect. 3: 655-677.
Mooi, F., I. H. Van Loo, M. van Gent, Q. He, M. J. Bart, K. J. Heuvelman, et al. 2009. Bordetella pertussis strains with increased toxin production associated with pertussis resurgence. Emerg. Infect. Dis. 15: 1206-1213.
Morbidity and Mortality Weekly Report. 2011. General Recommendations on Immunization. Recommendations of the Advisory Committee on Immunization Practices (ACIP) Recommendations and Reports. Vol. 60, No. 2.
Mukkur, T. K., G. H. McDowell, B. A. D. Stocker, and A. K. Lascelles. 1987. Protection against experimental salmonellosis in mice and sheep by immunisation with aromatic-dependent Salmonella Typhimurium. J. Med. Microbiol. 24: 11-19.
Nasso, M., G. Fedele, F. Spencieri, R. Palazzo, P. Costantino, R. Rappuoli, et al. 2011. Genetically detoxified pertussis induces Th1/Th17 immune response through MAPKs and IL-10dependent mechanisms. J. Immunol. 183: 1892-1899.
National Notifiable Diseases Surveillance System. 2011. Available at http://www9.health.gov.au/cda/Source/Rpt_3.cfm, accessed4/3/2011.
Olson, L. C. 1975. Pertussis. Medicine (Baltimore) 54: 427-469.
Parkhill, J., M. Sebaihia, A. Preston, L. D. Murphy, N. Thomson, D. E. Harris, et al. 2003. Comparative analysis of the genome sequences of Bordetella pertussis, Bordetella parapertussis and Bordetella bronchiseptica. Nat. Genet. 35: 32-40.
Pertussis [whooping cough]: Outbreaks. http://www.cdc.gov/pertussis/outbreaks.html. Accessed 14 June 2011.
Quinn, P., M. Gold, J. Royle, J. Buttery, J. P. Richmond, P. McIntyre, et al. 2011. Recurrence of extensive injection site reactions following DTPa or dTpa vaccine in children 406 years old. Vaccine 29: 4230-4237.
Rennels, M. B., M. A. Deloria, M. E. Pichichero, G. A. Losonsky, J. A. Englund, B. D. Meade, et al. 2000. Extensive swelling after booster doses of acellular pertussis-tetanusdiphtheria vaccines. Pediatrics 1: e1-e12.
Roberts, M., D. Maskell, P. Novotny, and G. Dougan. 1990. Construction and characterization in vivo of Bordetella pertussis aroA mutants. Infect. Immun. 58: 732-739.
Rowe, J., S. T. Yerkovich, P. Richmond, D. Suriyaarachchi, E. Fisher, L. Feddema, et al. 2005. Th2-associated local reactions to the acellular diphtheria-tetanus-pertussis vaccine in 4- to 6-yearold children. Infect. Immun. 73: 8130-8135.
Rossetti, T. R. 1997. Cloning the aroD gene of Bordetella pertussis. BSc. Hons. Thesis, University of Southern Queensland, Toowoomba, Queensland, Australia.
Sambrook, J., E. F. Fritsch, and Maniatis, T. 1989. Molecular Cloning: A Laboratory Manual, 2nd Ed. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY.
Schellekens, J., C. H. von Konig, and P. Gardner. 2005. Pertussis sources of infection and routes of transmission in the vaccination era. Pediatr. Infect. Dis. J. 24: 721-728.
Shahin, R., M. Leef, J. Eldridge, M. Hudson, and R. Gilley. 1995. Adjuvanticity and protective immunity elicited by Bordetella pertussis antigens encapsulated in poly[DL-lactide-co-glycolide] microspheres. Infect. Immun. 63: 1195-1200.
Stainer, D. W. and M. J. Scholte. 1971. A simple and chemically defined medium for the production of phase I Bordetella pertussis. J. Gen. Microbiol. 34: 778-784.
Towbin, H., T. Stachelin, and J. Gordon. 1979. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: Procedure and some applications. Proc. Natl. Acad. Sci. USA 76: 4350-4354.
Verma, N. K. and A. A. Lindberg. 1991. Construction of aromatic dependent Shigella flexneri 2a live vaccine candidate strains: Deletion mutations in the aroA and the aroD genes. Vaccine 9: 6-9.
Wearing, H. J. and P. Rohani. 2009. Estimating the duration of pertussis immunity using epidemiological signatures. PLoS Pathog. 5: e1000647.
Weiss, A. A. and E. L. Hewlett. 1986. Virulence factors of Bordetella pertussis. Annu. Rev. Microbiol. 40: 661-686.
Wendelboe, A. M., E. Njamkepo, A. Bourillon, D. D. Floret, J. Gaudelus, M. Gerber, et al. 2007. Infant Pertussis Study Group. Transmission of Bordetella pertussis to young infants. Pediatr. Infect. Dis. J. 26: 293-299.
Rieber, N., A. Graf, D. Hartl, S. Urschel, B. H. Belohradsky, and J. Liese. 2011. Acellular pertussis booster in adolescents induces Th1 and memory CD8+ T cell immune response. PLoS ONE 6: e17271.