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Applied and Environmental Microbiology, September 2002, p. 4253-4258, Vol. 68, No. 9
0099-2240/02/$04.00+0     DOI: 10.1128/AEM.68.9.4253-4258.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Purification and Immunogenicity of a Recombinant Bordetella pertussis S1S3FHA Fusion Protein Expressed by Streptococcus gordonii

Song F. Lee,1,2* Scott A. Halperin,2,3 Jennifer B. Knight,2 and Aaron Tait1

Department of Applied Oral Sciences, Faculty of Dentistry,1 Department of Microbiology & Immunology,2 Department of Pediatrics, Dalhousie University, Halifax, Nova Scotia, Canada B3H 3J53

Received 4 March 2002/ Accepted 31 May 2002

Acellular pertussis vaccines typically consist of antigens isolated from Bordetella pertussis, and pertussis toxin (PT) and filamentous hemagglutinin (FHA) are two prominent components. One of the disadvantages of a multiple-component vaccine is the cost associated with the production of the individual components. In this study, we constructed an in-frame fusion protein consisting of PT fragments (179 amino acids of PT subunit S1 and 180 amino acids of PT subunit S3) and a 456-amino-acid type I domain of FHA. The fusion protein was expressed by the commensal oral bacterium Streptococcus gordonii. The fusion protein was secreted into the culture medium as an expected 155-kDa protein, which was recognized by a polyclonal anti-PT antibody, a monoclonal anti-S1 antibody, and a monoclonal anti-FHA antibody. The fusion protein was purified from the culture supernatant by affinity and gel permeation chromatography. The immunogenicity of the purified fusion protein was assessed in BALB/c mice by performing parenteral and mucosal immunization experiments. When given parenterally, the fusion protein elicited a very strong antibody titer against the FHA type I domain, a moderate titer against native FHA, and a weak titer against PT. When given mucosally, it elicited a systemic response and a mucosal response to FHA and PT. In Western blots, the immune sera recognized the S1, S3, and S2 subunits of PT. These data collectively indicate that fragments of the pertussis vaccine components can be expressed in a single fusion protein by S. gordonii and that the fusion protein is immunogenic. This multivalent fusion protein approach may be used in designing a new generation of acellular pertussis vaccines.

* Corresponding author. Mailing address: Department of Applied Oral Sciences, Faculty of Dentistry, Dalhousie University, Halifax, Nova Scotia, Canada B3H 3J5. Phone: (902) 494-8799. Fax: (902) 494-6621. E-mail: Song.Lee{at}Dal.ca.

Applied and Environmental Microbiology, September 2002, p. 4253-4258, Vol. 68, No. 9
0099-2240/02/$04.00+0     DOI: 10.1128/AEM.68.9.4253-4258.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Knight, J. B., Halperin, S. A., West, K. A., Lee, S. F. (2008). Expression of a Functional Single-Chain Variable-Fragment Antibody against Complement Receptor 1 in Streptococcus gordonii. CVI 15: 925-931 [Abstract] [Full Text]  
  • Hussein, A. H., Davis, E. M., Halperin, S. A., Lee, S. F. (2007). Construction and Characterization of Single-Chain Variable Fragment Antibodies Directed against the Bordetella pertussis Surface Adhesins Filamentous Hemagglutinin and Pertactin. Infect. Immun. 75: 5476-5482 [Abstract] [Full Text]  
  • Lee, C. W., Lee, S. F., Halperin, S. A. (2004). Expression and Immunogenicity of a Recombinant Diphtheria Toxin Fragment A in Streptococcus gordonii. Appl. Environ. Microbiol. 70: 4569-4574 [Abstract] [Full Text]  
  • Kamachi, K., Arakawa, Y. (2004). Expression of a C Terminally Truncated Form of Pertussis Toxin S1 Subunit Effectively Induces Protection against Pertussis Toxin following DNA-Based Immunization. Infect. Immun. 72: 4293-4296 [Abstract] [Full Text]  
  • Lee, S. F., Halperin, S. A., Salloum, D. F., MacMillan, A., Morris, A. (2003). Mucosal Immunization with a Genetically Engineered Pertussis Toxin S1 Fragment-Cholera Toxin Subunit B Chimeric Protein. Infect. Immun. 71: 2272-2275 [Abstract] [Full Text]  


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