Induction of neutralizing antibodies in mice immunized with an amino-terminal polypeptide of Streptococcus mutans P1 protein produced by a recombinant Bacillus subtilis strain

Milene B. Tavares, Bruno M. Silva, Rafael C.M. Cavalcante, Renata D. Souza, Wilson B. Luiz, Juliano D. Paccez, Paula J. Crowley, L. Jeannine Brady, Luís C.S. Ferreira, Rita C.C. Ferreira

Research output: Contribution to journalArticlepeer-review

7 Scopus citations

Abstract

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 (P139-512) derived from the S. mutans strain UA159. Purified P139-512 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 P139-512 induced antibodies that reacted specifically with native P1 on the surface of S. mutans cells. The anti-P139-512 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.

Original languageEnglish (US)
Pages (from-to)131-142
Number of pages12
JournalFEMS Immunology and Medical Microbiology
Volume59
Issue number2
DOIs
StatePublished - Jul 2010

Keywords

  • Antibody responses
  • Bacillus subtilis
  • P1 protein
  • Recombinant proteins
  • Saliva-binding region
  • Streptococcus mutans

ASJC Scopus subject areas

  • Immunology and Allergy
  • Microbiology
  • Immunology
  • Microbiology (medical)
  • Infectious Diseases

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