ミュータンス群レンサ球菌 (S. sobrinus, S. downei) の共通抗原に対する単クロン抗体 : 特異性と抗原決定基の解析

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Antibody producing hybridomas were prepared by fusing immune spleen cells of a BALB/c mouse with myeloma cells. Nine hybridomas were isolated and their culture fluids were examined by enzyme immunoassay for reactivity with various strains of S. cricetus (serotype a), S. rattus (serotype b), S. mutans (serotype c, e, and f) S. sobrinus (serotype d and g), S. downei (serotype h), S. ferus (serotype c) and other streptococcal species predominantly found in the oral cavity. Ascitic fluids were collected from mice receiving one (d2-11) of the hybridoma clones, precipitated with ammonium sulphate, and purified a on DEAE-cellulose column. A polysaccharide antigen was extracted by autoclaving the cells of S. sobrinus (MT615R) in saline solution and purified on DEAE-Sephadex A-25, Sephadex G-200 and CM-Sephadex columns. The purified polysaccharide antigen consisted of rhamnose, galactose, and glucose, when determined by chemical analyses. Reactivity of purified monoclonal antibody (d2-11) was examined by enzyme immunoassay, precipitin reactions, and competitive precipitin inhibition tests. It was found in enzyme immunoassay using whole cells of microorganisms that the antibody reacted only with strains of S. sobrinus and S. downei and not those of many streptococcal species. It was also found in the competitive precipitin inhibition tests that β-galactpyranoside had a marked inhibitory effect while galactose and maltose had lesser effects. These results indicated that the β-galactosyl moiety is the structure involved in the predominant antigenic determinant of the cross-reactive polysaccharide antigen. Furthermore, the monoclonal antibody should be useful for a rapid identification of the streptococcal species among oral streptococci in dental plaque.
九州歯科学会の論文
1994-12-25

ミュータンス群レンサ球菌 (S. sobrinus, S. downei) の共通抗原に対する単クロン抗体 : 特異性と抗原決定基の解析

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