Previous Article | Next Article ![]()
Infection and Immunity, March 1999, p. 1511-1516, Vol. 67, No. 3
Department of Applied Oral Sciences,
Received 12 August 1998/Returned for modification 4 November
1998/Accepted 9 December 1998
In this study, the expression of the Bordetella
pertussis S1 subunit was tested in Streptococcus
gordonii, a commensal oral bacterium which has the potential to
be a live oral vaccine vehicle. The DNA fragment encoding the
N-terminal 179 amino acids of the S1 subunit was ligated into the
middle part of spaP, the surface protein antigen P1 gene
originating from Streptococcus mutans. The resulting
construct, carried on the Escherichia coli-Streptococcus shuttle vector pDL276, was introduced into S. gordonii DL-1
by natural transformation. One of the transformants (RJMIII) produced a
187-kDa protein (the predicted size of the SpaP-S1 fusion protein) which was recognized by both the anti-pertussis toxin (anti-PT) and
anti-SpaP antibodies, suggesting that an in-frame fusion had been made.
Results from immunogold-electron microscopic studies and cellular
fractionation studies showed that the fusion protein was surface
localized and was mainly associated with the cell wall of RJMIII,
indicating that SpaP was able to direct the fusion protein to the cell
surface. A rabbit antiserum raised against heat-killed S. gordonii RJMIII recognized the native S1 subunit of PT in Western
blotting and showed a weak neutralization titer to PT by the Chinese
hamster ovary cell-clustering assay. BALB/c mice immunized with the
heat-killed S. gordonii RJMIII were protected from the
toxic effect of PT in the leukocytosis-promoting and histamine
sensitization assays. In conclusion, a fragment of the S1 subunit of PT
was successfully surface expressed in S. gordonii; the
recombinant S1 fragment was found to be immunogenic and could induce
protection against the toxic effect of PT in mice.
0019-9567/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
Surface Expression of a Protective Recombinant
Pertussis Toxin S1 Subunit Fragment in Streptococcus
gordonii
*
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.
Infection and Immunity, March 1999, p. 1511-1516, Vol. 67, No. 3
0019-9567/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
This article has been cited by other articles:
| J. Bacteriol. | J. Virol. | Eukaryot. Cell |
|---|
| Microbiol. Mol. Biol. Rev. | Clin. Vaccine Immunol. | All ASM Journals |
|---|