Differential immunogenicity of a DNA vaccine containing the Streptococcus mutans wall-associated protein A gene versus that containing a truncated derivative antigen A lacking in the hydrophobic carboxyterminal region

DNA Cell Biol. 2005 Sep;24(9):574-81. doi: 10.1089/dna.2005.24.574.

Abstract

Two plasmid DNA constructs were obtained by cloning separately into the eukaryotic expression vector pcDNA3.1/V5-His-TOPO the wall-associated protein A (wapA) gene of Streptococcus mutans GS-5 or its truncated derivative antigen A (agA) gene encoding a known candidate antigen for dental caries vaccine. The immunogenicity of the two constructs, designated pcDNA-wapA and pcDNA-agA, was compared by intranasal immunization of two groups of mice using the cationic DMRIE-C (1,2-dimyristyloxypropyl-3-dimethylhydroxy ethyl ammonium bromide-cholesterol) as an adjuvant. Immunization with pcDNA-wapA or pcDNA- agA resulted in specific salivary IgA and systemic IgG antibodies to the target antigens after two doses given at 3-week intervals. Higher salivary IgA level was observed in the mice immunized with the pcDNA-wapA vaccine compared to those immunized with the pcDNA-agA vaccine. Furthermore, anti-WapA antibody inhibited S. mutans sucrose-dependent adherence suggesting a potential protection against S. mutans colonization of the tooth, while anti-AgA had no significant effect. Indeed, prediction and analysis of protein epitopes showed that WapA contains highly promiscuous MHC-II binding motifs in addition to those found in AgA. Immunodot assay confirmed that WapA bound biotin-labeled dextran, whereas AgA did not. These data indicated that full-length WapA is a better candidate vaccine antigen than the soluble AgA, which is truncated in the hydrophobic membrane and wall-spanning region.

Publication types

  • Comparative Study
  • Research Support, N.I.H., Extramural
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Antibodies, Anti-Idiotypic / immunology
  • Antigens, Bacterial / genetics*
  • Antigens, Bacterial / immunology*
  • Bacterial Adhesion / immunology
  • Bacterial Proteins / genetics*
  • Bacterial Proteins / immunology*
  • Biotin / metabolism
  • Dental Caries / prevention & control*
  • Dextrans / metabolism
  • Enzyme-Linked Immunosorbent Assay
  • Epitopes / genetics
  • Epitopes / metabolism
  • Escherichia coli
  • Female
  • Genes, MHC Class II / genetics
  • Immunoglobulin A / immunology
  • Immunoglobulin G / immunology
  • Lipids
  • Mice
  • Mice, Inbred BALB C
  • Plasmids / genetics
  • Quaternary Ammonium Compounds
  • Saliva / immunology
  • Vaccines, DNA / genetics
  • Vaccines, DNA / immunology*

Substances

  • Antibodies, Anti-Idiotypic
  • Antigens, Bacterial
  • Bacterial Proteins
  • Dextrans
  • Epitopes
  • Immunoglobulin A
  • Immunoglobulin G
  • Lipids
  • Quaternary Ammonium Compounds
  • Vaccines, DNA
  • wapA protein, Bacillus subtilis
  • (3-dimyristyloxypropyl)(dimethyl)(hydroxyethyl)ammonium
  • Biotin