Antigens of Streptococcus mutans: characterization of a polysaccharide antigen from walls of strain GS 5

Abstract

A cell wall associated polysaccharide antigen was isolated from S. mutans GS 5 and appeared to determine serotype c specificity. Ouchterlony double diffusion analysis of crude formamide extracts derived from purified cell walls of two serotype c strains (GS 5 and JC 2) showed complete identity when reacted with anti GS 5 sera. Immunoelectrophoresis of this extract demonstrated the typical mobility for this serotype as described by others. Column chromatography on BioGel P 100 of the crude formamide extracts derived from GS 5 walls resulted in a single antigenic peak being resolved. This material, when loaded onto a diethylaminoethylcellulose column and eluted with a linear gradient of ammonium carbonate (0.0 to 0.2 M), was resolved further into two serologically reactive peaks (I and II). Only two constituents, rhamnose and glucose, were detected in the purified column fractions. Peak I had a rhamnose to glucose molar ratio of 0.9:1.0, and peak II, the major resolvable fraction, had a molar ratio of 1.7:1.0. The peak II ratio was very similar to that found in the formamide extract residue pellet (1.6:1.0). Ouchterlony analysis of the crude formamide extract and the purified fractions revealed only partial identity between peaks I and II but complete identity between peak II and the crude extract. Likewise, immunoelectrophoresis showed no differences in mobility of peak II and the crude extract, whereas peak I moved towards the cathode. Possible structural relationships between the two antigenic fractions are discussed below. Hapten inhibition studies suggested that an α glucosyl group is at the immunodeterminant site of the antigen.