Induction of neutralizing antibodies in mice immunized with an amino-terminal polypeptide of Streptococcus mutans P1 protein produced by a recombinant Bacillus subtilis strain.
The oral pathogen Streptococcus mutans expresses a surface protein, P1, which interacts with the salivary pellicle on the tooth surface or with fluid-phase saliva, resulting in bacterial adhesion or aggregation, respectively. P1 is a target of protective immunity. Its N-terminal region has been associated with adhesion and aggregation functions and contains epitopes recognized by efficacious antibodies. In this study, we used Bacillus subtilis, a gram-positive expression host, to produce a recombinant N-terminal polypeptide of P1 (P1(39-512)) derived from the S. mutans strain UA159. Purified P1(39-512) reacted with an anti-full-length P1 antiserum as well as one raised against intact S. mutans cells, indicating preserved antigenicity. Immunization of mice with soluble and heat-denatured P1(39-512) induced antibodies that reacted specifically with native P1 on the surface of S. mutans cells. The anti-P1(39-512) antiserum was as effective at blocking saliva-mediated aggregation of S. mutans cells and better at blocking bacterial adhesion to saliva-coated plastic surfaces compared with the anti-full-length P1 antiserum. In addition, adsorption of the anti-P1 antiserum with P1(39-512) eliminated its ability to block the adhesion of S. mutans cells to abiotic surfaces. The present results indicate that P1(39-512), expressed and purified from a recombinant B. subtilis strain, maintains important immunological features of the native protein and represents an additional tool for the development of anticaries vaccines.
Tavares
Milene B
MB
Department of Microbiology, Institute of Biomedical Sciences, University of São Paulo, SP, Brazil.
Silva
Bruno M
BM
Cavalcante
Rafael C M
RC
Souza
Renata D
RD
Luiz
Wilson B
WB
Paccez
Juliano D
JD
Crowley
Paula J
PJ
Brady
L Jeannine
LJ
Ferreira
Luís C S
LC
Ferreira
Rita C C
RC
D018829:Adhesins, Bacterial / Q000235:genetics / Q000276:immunology*; D000818:Animals; D000907:Antibodies, Bacterial / Q000097:blood*; D057134:Antibodies, Neutralizing / Q000097:blood*; D001412:Bacillus subtilis / Q000235:genetics*; D001422:Bacterial Adhesion / Q000276:immunology; D005822:Genetic Vectors; D051379:Mice; D008807:Mice, Inbred BALB C; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology; D013295:Streptococcus mutans / Q000235:genetics / Q000276:immunology*
D018829:Adhesins, Bacterial; D000907:Antibodies, Bacterial; D057134:Antibodies, Neutralizing; D011994:Recombinant Proteins
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