Cheung A L, Fischetti V A
Laboratory of Bacteriology and Immunology, Rockefeller University, New York, New York 10021.
Infect Immun. 1988 May;56(5):1061-5. doi: 10.1128/iai.56.5.1061-1065.1988.
To evaluate the variation in the expression of cell wall antigens between Staphylococcus aureus grown in liquid medium and solid support, bacteria were harvested from liquid chemically defined medium and chemically defined medium in a 1% agar base. Cell wall proteins were then extracted by lysostaphin in a protoplast-stabilizing medium (30% raffinose). After separation of the cell wall antigens by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blots, they were probed with chicken antiserum to an S. aureus strain grown on a solid support. For each of the 15 clinical strains analyzed, high-molecular-size bands (molecular size range, 120 to 220 kilodaltons) were either enhanced or distinctly present when compared with those from the cell wall extract of the same strain grown in liquid medium. Results of enzymatic treatment of whole staphylococci grown on solid medium suggested the proteinaceous nature and the surface location of these antigens. Limited passage studies demonstrated the ability of the staphylococci to alter these surface proteins when passaged alternately on liquid and solid media. These observations suggested the importance of the microenvironment to the expression of cell wall proteins in S. aureus. Correlations with observations in vivo may help identify the determinants of microbial pathogenicity in S. aureus.
为评估在液体培养基和固体支持物上生长的金黄色葡萄球菌之间细胞壁抗原表达的差异,从液体化学限定培养基和含1%琼脂基质的化学限定培养基中收获细菌。然后在原生质体稳定培养基(30%棉子糖)中用溶葡萄球菌素提取细胞壁蛋白。通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳和蛋白质印迹分离细胞壁抗原后,用针对在固体支持物上生长的金黄色葡萄球菌菌株的鸡抗血清进行检测。对于所分析的15株临床菌株中的每一株,与在液体培养基中生长的同一菌株的细胞壁提取物相比,高分子量条带(分子大小范围为120至220千道尔顿)要么增强,要么明显出现。对在固体培养基上生长的全葡萄球菌进行酶处理的结果表明这些抗原具有蛋白质性质且位于表面。有限传代研究表明,当在液体和固体培养基上交替传代时,葡萄球菌能够改变这些表面蛋白。这些观察结果表明微环境对金黄色葡萄球菌细胞壁蛋白表达的重要性。与体内观察结果的相关性可能有助于确定金黄色葡萄球菌中微生物致病性的决定因素。