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Applied and Environmental Microbiology, August 2004, p. 4569-4574, Vol. 70, No. 8
0099-2240/04/$08.00+0     DOI: 10.1128/AEM.70.8.4569-4574.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Expression and Immunogenicity of a Recombinant Diphtheria Toxin Fragment A in Streptococcus gordonii

Chiang W. Lee,1,2,3,4,{dagger} Song F. Lee,1,2,4* and Scott A. Halperin2,3,4

Department of Applied Oral Sciences, Faculty of Dentistry,1 Department of Microbiology and Immunology,2 Department of Pediatrics, Faculty of Medicine, Dalhousie University,3 Izaak Walton Killam Health Centre, Halifax, Nova Scotia B3H 3J5, Canada4

Received 3 November 2003/ Accepted 13 April 2004

A nontoxic mutant diphtheria toxin fragment A (DTA) was genetically fused in single, double, or triple copy to the major surface protein antigen P1 (SpaP) and surface expressed in Streptococcus gordonii DL-1. The expression was verified by Western immunoblotting. Mouse antisera raised against the recombinant S. gordonii recognized the native diphtheria toxinm suggesting the recombinant DTA was immunogenic. When given intranasally to mice with cholera toxin subunit B as the adjuvant, the recombinant S. gordonii expressing double copies of DTA (SpaP-DTA2) induced a mucosal immunoglobulin A response and a weak systemic immunoglobulin G response. S. gordonii SpaP-DTA2 was able to orally colonize BALB/c mice for a 15-week period and elicited a mucosal response, but a serum immunoglobulin G response was not apparent. The antisera failed to neutralize diphtheria toxin cytotoxicity in a Vero cell assay.

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

{dagger} Present address: Department of Microbiology and Immunology, College of Veterinary Medicine, Cornell University, Ithaca, NY 14853-6401.

Applied and Environmental Microbiology, August 2004, p. 4569-4574, Vol. 70, No. 8
0099-2240/04/$08.00+0     DOI: 10.1128/AEM.70.8.4569-4574.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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