Cross-reactions of Streptococcus mutans due to cell wall teichoic acid

Hot saline extracts of Streptococcus mutans have been shown to contain antigenic substances which occasionally react nonspecifically with some antisera against whole cells of various serological groups and types of streptococci. Chromatography of the extract of S. mutans strain MT703 (serotype e) on a diethylaminoethyl-Sephadex A-25 column gave two principal antigens. One antigen was eluted without adsorption to the resin and was identified as the serotype-specific polysaccharide. The other antigen, which contained a large quantity of phosphorus, was adsorbed to and released from the resin by gradient elution. It was reactive against the antisera specific for polyglycerophosphate (PGP) from group A Streptococcus pyogenes and/or S. mutans strain Ingbritt (type c). The PGP antigen was further purified by gel filtration with Sephadex G-75. Two peaks, PGP-1 and PGP-2, were obtained. Each possessed the same antigenic specificity to anti-PGP serum as shown by immunodiffusion. Chemical analyses revealed that the molar ratio of phosphorus to glycerol in both was about 1:1, although the protein content between the two was significantly different. PGP antigen was found to be widely distributed in hot saline extracts from various gram-positive bacteria, with a few exceptions. However, all gramnegative bacteria examined were free of PGP. The PGP in the hot saline extracts of various gram-positive bacteria possessed an essentially identical antigenic specificity. The addition of diethylaminoethyl-Sephadex A-25 resin to hot saline extracts successfully removed the cross-reacting PGP antigen. After adsorption of the extract from S. mutans, the supernatant contained only type-specific polysaccharide antigen, except type b, in which both type b-specific polysaccharide and PGP antigens were adsorbed with the resin. This simple procedure should be useful for the removal of the PGP-type teichoic acid from antigen extracts of bacteria that contain uncharged polysaccharides. Umemoto (New York State University School of Dentistry at Buffalo, Buffalo), B. Guggenheim 903 on July 15, 2020 by guest http://iai.asm.org/ Downloaded from

Section: Downloaded Fromsupporting

confidence: 71%

Section: Discussionmentioning

confidence: 99%

Selective adsorption of heterophile polyglycerophosphate antigen from antigen extracts of Streptococcus mutans and other gram-positive bacteria

Hamada¹,

Tai²,

Slade³

1976

“…The second phosphorus peak showed no serological activity, and the conclusion was drawn that it was nucleic acid in nature. PGP was found to be the cross-reacting antigen among many gram-positive bacteria (6,21,36), including S. mutans (4,16,18,27). Recently, we have found that the addition of DEAE-Sephadex resin to a crude antigen extract selectively adsorbed the cross-reacting PGP without de-tectable reduction of its antigenicity (Hamada, Tai, and Slade, unpublished data).…”

Section: Discussionmentioning

confidence: 99%

Purification and immunochemical characterization of type e polysaccharide antigen of Streptococcus mutans

Hamada¹,

Slade²

1976

The type-specific antigen of Streptococcus mutans strain MT703, serotype e, has been chromatographically purified and characterized. Two chromatographic fractions were obtained from saline extracts which reacted with both anti-MT703 whole-cell serum and Lancefield group E serum. The major fraction (eI) was identified as a polysaccharide composed of 37% glucose, 56% rhamnose, 5% protein, and 0.3% phosphorus, whereas the minor fraction (eII) contained 66% protein in addition to 10% glucose and 17% rhamnose. The immunological specificity of these antigens was found to be the same by immunodiffusion in agar gel. Another fraction with a negative charge (eIII) reacted with polyglycerophosphate antisera from Streptococcus mutans and Streptococcus pyogenes. For comparison, the MT703 antigen in a hot trichloroacetic acid extract (eA) and the group E antigen from a saline extract of cells of strain K129 (EI) were similarly purified by anionic ion-exchange chromatography. Although the ratio of glucose and rhamnose in eA was 1:0.9 and in eI and eII approximately 1:1.5, reactions of identity were obtained in gel diffusion against specific anti-e serum. This difference in ratio is probably a result of the extraction procedures. Both the type e and group E antisera were reactive with both eI and EI antigens. The adsorption of group E antiserum with MT703 cells removed all E antibody, whereas type e-specific antibody remained after adsorption with K129 cells. These results suggest that eI antigen possesses both e and E specificities, whereas EI possesses E only. These findings were supported by the quantitative precipitin test and immunodiffusion and/or immunoelectrophoretic patterns in agar gel. Methyl-,-i->glucopyranoside markedly inhibited the precipitin reaction in both type e and group E sera. However, a significantly stronger inhibition by cellobiose of type e serum than of group E serum indicates that a ,-linked glucose-glucose dimer is the predominant antigenic determinant of the e specificity. The presence of both e and E specificities on a single polysaccharide molecule was demonstrated by the use of purified e antigen released from a specific e-anti-e complex. This antigen reacted with a group E-specific serum as well as a type e-specific serum. An examination of five S. mutans type e strains showed the presence of group E specificity also, whereas the I, II, and IV serotypes of group E streptococci only possessed the group E specificity. Strains of Streptococcus mutans, an agent that develops dental caries in animals and probably humans, have been subdivided into seven immunological types (2, 30). The antigens of types a (27), b (26), c (16, 42), d (13, 18) and f (8a) are located in the cell wall and are polysaccharides composed of rhamnose and glucose (type c and f), rhamnose and galactose (type b), and glucose and galactose (types a and d). S. mutans strains do not show any immunological cross-reaction with the streptococcal groups except group E (3, 30).Bratthall (3) considered that type e S. mutans possessed the ...

“…described elsewhere (2). Rabbit RBC suspensions optimally coated with a standard GTA preparation were then added, incubated, and read for PHG inhibition.…”

mentioning

confidence: 99%

Lipid-free glycerol teichoic acids with potent membrane-binding activity

Cooper¹,

Chorpenning²,

Roses³

1978

Self Cite

Lipid analysis of several glycerol teichoic acid preparations strongly indicated that covalently bound lipid is not required for spontaneous adsorption of glycerol teichoic acid to erythrocyte membranes. Although fatty acids were detected in each of four batches, none were covalently bound. Chloroform-ether-extracted antigens retained potent erythrocyte membrane-binding activity as measured by passive hemagglutination, even though they were shown to contain less than one fatty acid residue per 4,869 teichoic acid chains. Mild ammonolysis abolished erythrocyte-sensitizing activity in passive hemagglutination, but further studies indicated the loss of activity was due to partial destruction of the polyglycerophosphate backbone and not to the removal of esterified lipid. The amount of hydrolyzed antigen required to produce 100% passive hemagglutination inhibition was between 170 and 330 times the amount required to produce the same result using unhydrolyzed glycerol teichoic acid. The average chain length was reduced from 19.1 to 9.7, 7.4, and 5.1 glycerophosphate residues for antigen samples hydrolyzed for 1, 5, and 16 h, respectively.