title: Immunogenic and protective potential of mutans streptococcal glucosyltransferase peptide constructs selected by major histocompatibility complex class II allele binding.
abstract: Mutans streptococcal glucosyltransferases (GTF) have been demonstrated to be effective components of dental caries vaccines. We had previously selected peptide subunits of GTF for vaccine development based on putative functional significance and conservation of GTF primary structure among enzyme isoforms. In this study, 20 20-mer linear GTF peptides were synthesized, 17 identified on the basis of the highest potential major histocompatibility complex (MHC) class II-binding activity using computer-generated algorithms (Epimatrix and ProPred) and 3 with previously demonstrated functional significance. The immunoreactivities of these peptides were explored with rodent systems. Sera from GTF-immunized rats, assessed for binding to linear peptides by enzyme-linked immunosorbent assay, demonstrated immunoglobulin G antibody reactivity with peptides 6 and 11 and a T-cell proliferation response to peptides 6, 9, 11, and 16. Multiple antigenic peptide (MAP) constructs were synthesized from promising linear sequences. Rats that were immunized with MAP 7, 11, or 16, respectively, responded well to the immunizing MAP. Most importantly, a robust immune response (antibody and T-cell proliferation) was observed to native GTF following MAP 11 (amino acids 847 to 866; VVINNDKFVSWGITDFEM) immunization. This response inhibited GTF enzyme function. Two dental caries pathogenesis experiments were performed wherein rats were immunized with MAP constructs 11, 16, and/or 11 plus 16, followed by infection with cariogenic Streptococcus sobrinus. In both experiments cariogenic bacterial recoveries were reduced relative to total streptococci in the MAP 11- and MAP 11 plus 16-immunized groups, and the extent of dental caries was also significantly reduced in these groups. Thus, we have identified a peptide with projected avid MHC-binding activity that elicited immunoreactivity with native GTF and demonstrated protection against dental caries infection after immunization, implying that this peptide may be important in a subunit dental caries vaccine.
authors.affiliation: Department of Immunology, The Forsyth Institute, 140 The Fenway, Boston, MA 02115-3799, USA.
authors.forename: S
authors.initials: S
authors.lastname: Culshaw
authors.forename: K
authors.initials: K
authors.lastname: Larosa
authors.forename: H
authors.initials: H
authors.lastname: Tolani
authors.forename: X
authors.initials: X
authors.lastname: Han
authors.forename: J W
authors.initials: JW
authors.lastname: Eastcott
authors.forename: D J
authors.initials: DJ
authors.lastname: Smith
authors.forename: M A
authors.initials: MA
authors.lastname: Taubman
mesh_terms: D000818:Animals; D000907:Antibodies, Bacterial / Q000097:blood; D049109:Cell Proliferation; D015169:Colony Count, Microbial; D019295:Computational Biology / Q000379:methods; D004195:Disease Models, Animal; D004797:Enzyme-Linked Immunosorbent Assay; D005964:Glucosyltransferases / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000276:immunology*; D000949:Histocompatibility Antigens Class II / Q000276:immunology* / Q000378:metabolism; D007074:Immunoglobulin G / Q000097:blood; D008214:Lymphocytes / Q000276:immunology; D009055:Mouth / Q000382:microbiology; D009154:Mutation; D010455:Peptides / Q000138:chemical synthesis / Q000276:immunology*; D051381:Rats; D017207:Rats, Sprague-Dawley; D013290:Streptococcal Infections / Q000276:immunology*; D013291:Streptococcus / Q000145:classification / Q000302:isolation & purification; D016986:Streptococcus sobrinus / Q000201:enzymology / Q000276:immunology*; D022223:Vaccines, Subunit / Q000008:administration & dosage / Q000276:immunology*
chemical_list: D000907:Antibodies, Bacterial; D000949:Histocompatibility Antigens Class II; D007074:Immunoglobulin G; D010455:Peptides; D022223:Vaccines, Subunit; D005964:Glucosyltransferases