Forsyth Dental Infirmary for Children

The Forsyth Institute

The Henry M. Jackson Foundation for the Advancement of Military Medicine Inc.

Daniel J Smith

Martin A Taubman

Andrew Lees

Immunogenic compositions and subunit vaccines for dental caries are described which comprise peptide subunits of glucan binding protein-B and peptide subunits of glucan binding protein-B in combination with peptide subunits of glucosyltransferase. Methods of provoking an immune response toglucan binding protein-B or glucosyltransferase. Methods of immunizing a mammal against dental caries are also described, along with antibodies which bind particular epitopes of glucan binding protein-B or glucosyltransferase.

Immunogenicity of glucan binding protein

Glucan binding protein and glucosyltransferase immunogens

The present invention provides glucan-based compositions and methods for stimulating an immune response against mutans streptococci components and vaccines and methods for the treatment and prevention of dental caries. In a preferred embodiment, a glucan polymer, preferably WSG, is covalently bound to one or more T cell-dependent antigens to form a conjugate vaccine. The T cell-dependent antigen preferably contains epitopes of one or more mutans streptococcal proteins, such as a glucosyltransferase. Moreover, one or more moieties, including haptens, may be conjugated to the glucan-T cell-dependent composition. In a preferred embodiment, these moieties are peptides which contain immunogenic epitopes corresponding to components of a mutans streptococcus.

Conjugate vaccines for the prevention of dental caries

Vaccine compositions and immunogenic compositions are described which are glucosyltransferase subunit vaccines for dental caries and which contain at least one peptide which corresponds to a sequence of glucosyltransferase containing aspartate 413, an equivalent of aspartate 413, aspartate 451, an equivalent of aspartate 451, aspartate 562, an equivalent of aspartate 562, aspartate 567, an equivalent of aspartate 567, histidine 561, an equivalent of histidine 561, tryptophan 491, an equivalent of tryptophan 491, glutamate 489, an equivalent of glutamate 489, arginine 449, an equivalent of arginine 449, or combinations thereof. These subunit vaccines elicit antibodies which protect an immunized mammal from dental caries. Methods of provoking an immune response to intact glucosyltransferase are also described.

Glucosyltransferase immunogens

Vaccine compositions and immunogenic compositions are described which are glucosyltransferase subunit vaccines for dental caries and which contain at least one peptide which corresponds to a sequence of glucosyltransferase containing aspartate 413, an equivalent of aspartate 413, aspartate 451, an equivalent of aspartate 451, aspartate 562, and equivalent of aspartate 562, aspartate 567, an equivalent of aspartate 567, histidine 561, an equivalent of histidine 561, tryptophan 491, an equivalent of tryptophan 491, glutamate 489, an equivalent of glutamate 489, arginine 449, an equivalent of arginine 449, or combinations thereof. These subunit vaccines elicit antibodies which protect an immunized mammal from dental caries. Methods of provoking an immune response to intact glucosyltransferase are also described.

Synthetic peptide vaccines for dental caries

Immunization of animals with a composition containing either an amino acid sequence from the catalytic domain of glucosyltransferase, an amino acid sequence from the glucan-binding region of glucosyltransferase or an amino acid sequence from the native surface domain of glucosyltransferase provoke antibody and T-cell immune responses to this enzyme. Since this enzyme has been implicated in the colonization of mutans streptococci on tooth surfaces, such immune responses are important for the prevention of dental caries. Multicomponent and multivalent compositions which include these amino acid sequences provide effective vaccine capabilities.

A method of preparing a purified glucosyltransferase (GTF) for use in immunization against dental caries, which method comprises: culturing a Streptococcus mutans in a medium containing glucose and dialyzable nutrients to form a mixture of culture cells and supernatant; recovering the supernatant by the removal of the culture cells; admixing the recovered supernatant with a water-insoluble, polymerized polysaccharide as solid particulate material for a period of time, to provide a GTF, solid particulate complex; recovering the GTF complex in solid particulate form by filtration; washing the solid GTF complex to remove unbound GTF and medium components; removing GTF from the solid particulate material by a denaturing solvent; recovering the water-insoluble particulate material for reuse in the method; and recovering the GTF from the water-insoluble polysaccharide and purifying the recovered GTF.

Method for the preparation of glucosylranferase

Immunization of animals with preparations containing more purified forms of glucosyltransferase (GTF) results in the presence of antibody in saliva demonstrable by functional inhibitions of enzyme activity and binding of radioactive enzyme. Serum antibody was also present. Immunized groups of animals had lower mean caries scores than comparably sham-immunized or nonimmunized control groups. Local immunization with GTF of serotype c or g of a Streptococcus mutans reduces the colonization, caries, and lesions caused by infection with S. mutans of serotype g (strain 6715) or c, or with serotype g or c, or with serotype a or g, respectively.

Method of preparing a purified glucosyltransferase

Immunization of animals with preparations containing more purified forms of glucosyltransferase (GTF) results in the presence of antibody in saliva demonstrable by functional inhibitions of enzyme activity and binding of radioactive enzyme. Serum antibody was also present. Immunized groups of animals had lower mean caries scores than comparably sham-immunized or nonimmunized control groups. Local immunization with GTF of serotype c or g or a Streptococcus mutans reduces the colonization, caries, and lesions caused by infection with S. mutans of serotype g (strain 6715) or c, or with serotype g or c, or with serotype a or g, respectively.

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Natick, MA, United States of America

Daniel J Smith