title: Diepitopic construct of functionally and epitopically complementary peptides enhances immunogenicity, reactivity with glucosyltransferase, and protection from dental caries.
abstract: Coimmunization with peptide constructs from catalytic (CAT) and glucan-binding (GLU) domains of glucosyltransferase (GTF) of mutans streptococci has resulted in enhanced levels of antibody to the CAT construct and to GTF. We designed and synthesized a diepitopic construct (CAT-GLU) containing two copies of both CAT (B epitope only) and GLU (B and T epitope) peptides. The immunogenicity of this diepitopic construct was compared with that of individual CAT and GLU constructs by immunizing groups of Sprague-Dawley rats subcutaneously in the salivary gland vicinity with the CAT-GLU, CAT, or GLU construct or by treating rats by sham immunization. Levels of serum immunoglobulin G (IgG) antibody to GTF or CAT in the CAT-GLU group were significantly greater than in GLU- or CAT-immunized groups. Immunization with CAT-GLU was compared to coimmunization with a mixture of CAT and GLU in a second rodent experiment under a similar protocol. CAT-GLU immunization resulted in serum IgG and salivary IgA responses to GTF and CAT which were greater than after coimmunization. Immunization with the diepitopic construct and communization with CAT and GLU constructs showed proliferation of T lymphocytes to GTF. Immunization with either the CAT or GLU construct has been shown to elicit significant protection in a rodent dental caries model. Similarly in this study, the enhanced response to GTF after immunization with the CAT-GLU construct resulted in protective effects on dental caries. Therefore, the CAT-GLU diepitopic construct can be a potentially important antigen for a caries vaccine, giving rise to greater immune response than after immunization with CAT, GLU, or a mixture of the two.
authors.affiliation: Department of Immunology, The Forsyth Institute, Boston, Massachusetts 02115, USA. mtaubman@forsyth.org
authors.forename: M A
authors.initials: MA
authors.lastname: Taubman
authors.forename: C J
authors.initials: CJ
authors.lastname: Holmberg
authors.forename: D J
authors.initials: DJ
authors.lastname: Smith
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D020134:Catalytic Domain; D002455:Cell Division; D003731:Dental Caries / Q000517:prevention & control*; D018985:Epitopes, B-Lymphocyte / Q000276:immunology*; D018984:Epitopes, T-Lymphocyte / Q000276:immunology*; D005936:Glucans / Q000378:metabolism; D005964:Glucosyltransferases / Q000276:immunology*; D007070:Immunoglobulin A / Q000276:immunology; D007074:Immunoglobulin G / Q000097:blood / Q000276:immunology; D008969:Molecular Sequence Data; D010455:Peptides; D051381:Rats; D017207:Rats, Sprague-Dawley; D012469:Salivary Glands / Q000276:immunology; D016986:Streptococcus sobrinus / Q000201:enzymology; D013601:T-Lymphocytes / Q000276:immunology; D014611:Vaccination
chemical_list: D018985:Epitopes, B-Lymphocyte; D018984:Epitopes, T-Lymphocyte; D005936:Glucans; D007070:Immunoglobulin A; D007074:Immunoglobulin G; D010455:Peptides; D005964:Glucosyltransferases