Functional and immunogenic characterization of two cloned regions of Streptococcus mutans glucosyltransferase I

被引:30
作者
Jespersgaard, C
Hajishengallis, G
Greenway, TE
Smith, DJ
Russell, MW
Michalek, SM
机构
[1] Univ Alabama, Dept Microbiol, Birmingham, AL 35294 USA
[2] Univ Alabama, Dept Oral Biol, Birmingham, AL 35294 USA
[3] Forsyth Dent Ctr, Dept Immunol, Boston, MA 02115 USA
关键词
D O I
10.1128/IAI.67.2.810-816.1999
中图分类号
R392 [医学免疫学]; Q939.91 [免疫学];
学科分类号
100102 ;
摘要
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,,ve 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.
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收藏
页码:810 / 816
页数:7
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