ミュータンス群レンサ球菌 (Streptococcus rattus) に関する研究 : 抗血清による特異抗原の化学的, 免疫化学的性状とその細胞局在性

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  • Studies on the Mutans Group of Streptococci (Streptococcus rattus) : Chemical and Immunochemical Characterization of Specific Antigen and Its Localization by Use of Specific Antiserum

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A specific antiserum was prepared by immunizing a rabbit with whole cells of Streptococcus rattus strain FA1 and absorption with heterologous strains of the mutans group of streptococci. The absorbed antiserum was tested for its specificity in double diffusion, immunoelectrophoresis enzyme and radiommunoassay with whole cells of the mutans group of streptococci and other oral streptococcal species. Antigens were extracted from FA1, BHT (reference strains) and KAY1 (an isolate from dental plaque of a female student) strains, and purified on ion-exchange and gel chromatographic columns. The crude and purified antigens were analyzed chemically as well as immunologically by use of the antiserum for characterization of the antigenic determinants. The immunological reactions included the double diffusion, competitive precipitin reaction in the presence of haptenic sugars, immunoelectrophoresis and immunoblotting. The purified antigen consisted of protein, sugar, lipid and phosphorus. The antigenic determinant was resistant against digestion with proteinases, lipases, RNase and DNase. When EIA microtiter plates were coated with the purified antigen and treated with NaIO_4, reactivity with the antiserum diminished, demonstrating involvement of sugar moiety of the antigen. This was supported by the negative staining of the antigen when it was reacted with the antiserum after SDS-PAGE analysis. Gas chromatographic analysis revealed that the purified antigen contained a substantial amount of acidic and hydrophobic amino acids. Immunoelectron microscopy using ferritin-labelled anti-rabbit IgG was conducted to examine localization of the specific antigen. It was demonstrated that the antigen appeared as irregular masses localizing on the outer but not inner surface of the cell wall.

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