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Infection and Immunity, February 1999, p. 810-816, Vol. 67, No. 2
Departments of
Microbiology1 and
Oral
Biology,2 University of Alabama at Birmingham,
Birmingham, Alabama 35294, and
Department of Immunology,
Forsyth Dental Center, Boston, Massachusetts
021153
Received 31 July 1998/Returned for modification 16 September
1998/Accepted 12 November 1998
Glucosyltransferase (GTF) enzymes of mutans streptococci are
considered virulence factors due to their ability to synthesize adhesive glucans, which facilitate cell-to-cell adherence and accumulation. In this study we report the cloning, expression, and
characterization of the catalytic (CAT) and glucan-binding (GLU)
domains of S. mutans GTF-I encoded by gtfB. The
CAT and GLU polypeptides represent amino acid residues 253 to 628 and 1183 to 1473, respectively, of S. mutans GTF-I. Antibodies
to recombinant CAT and GLU were generated in rabbits and purified by
affinity chromatography. Purified anti-CAT antibodies significantly inhibited water-insoluble glucan synthesis by S. mutans and
S. sobrinus GTFs (P < 0.0001 and
P < 0.05, respectively). The purified anti-GLU
antibodies significantly inhibited both water-insoluble and
water-soluble glucan synthesis by S. mutans GTFs
(P < 0.0001 and P < 0.05, respectively). These results demonstrate that anti-CAT and anti-GLU
antibodies are capable of inhibiting a variety of GTF activities. Since
antibodies to S. mutans in saliva are implicated in
protection against disease, we next assessed the ability of CAT and GLU
polypeptides to induce mucosal antibody responses in mice. Intranasal
(i.n.) immunization of mice with CAT showed significantly
(P < 0.005) elevated levels of specific
immunoglobulin G (IgG) antibody activity in serum and specific IgA
antibody activity in serum, saliva, vaginal washes, and fecal samples.
GLU immunized animals showed significantly (P < 0.005) elevated levels of specific IgA antibody activity in serum and
vaginal secretions. Taken together, these results demonstrate that the
recombinant CAT and GLU polypeptides are effective in inducing both
mucosal and systemic immune responses. The ability of these
polypeptides to induce a mucosal IgA immune response in mice after i.n.
immunization supports their use as subunit vaccine candidates in the
development of an anticaries vaccine.
0019-9567/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
Functional and Immunogenic Characterization of Two
Cloned Regions of Streptococcus mutans
Glucosyltransferase I
*
Corresponding author. Mailing address: Department of
Microbiology, University of Alabama at Birmingham, 845 South 19th, BBRB 258, Birmingham, AL 35294-2170. Phone: (205) 934-3470. Fax: (205) 934-1426. E-mail: suemich{at}uab.edu.
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