McDermid K P, Morck D W, Olson M E, Dasgupta M K, Costerton J W
Department of Biological Sciences, University of Calgary, Alberta, Canada.
Infect Immun. 1993 May;61(5):1743-9. doi: 10.1128/iai.61.5.1743-1749.1993.
Staphylococcus epidermidis RP62A (ATCC 35984) was grown in tryptic soy broth (TSB), iron-depleted TSB (TSB-Fe), iron-reconstituted TSB-Fe (TSB+Fe), a chemically defined medium, and fetal calf serum (FCS) and on silastic disks in chambers that were sutured to the pig peritoneal wall. Bacterial cell wall proteins were extracted by digestion with recombinant lysostaphin, separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and detected by silver staining. Cell wall proteins from TSB-, chemically defined medium-, or FCS-grown cells had a complex profile of greater than 25 protein bands spanning the full molecular mass range. By contrast, a digest obtained from in vivo-grown cells had only five major proteins of 40 kDa or greater. Proteins of 130 and 106 kDa were present in the cell envelopes of TSB-Fe- and in vivo-grown cells but not in those grown in TSB or TSB+Fe. A 43-kDa protein expressed by in vitro-grown cells and 52- and 96-kDa proteins expressed by in vivo-grown cells reacted with antisera from pigs with the chamber implants and from catheterized, paracatheter-inoculated pigs but not with hyperimmune sera from pigs immunized with TSB-grown cells. The data indicate that S. epidermidis, growing under in vivo conditions, expresses antigens distinct from those that are grown in vitro.
表皮葡萄球菌RP62A（ATCC 35984）在胰蛋白胨大豆肉汤（TSB）、缺铁TSB（TSB-Fe）、铁重构TSB-Fe（TSB+Fe）、化学限定培养基和胎牛血清（FCS）中培养，并在缝合于猪腹膜壁的小室中的硅橡胶盘上培养。通过用重组溶葡萄球菌素消化提取细菌细胞壁蛋白，经十二烷基硫酸钠-聚丙烯酰胺凝胶电泳分离，并用银染法检测。来自TSB、化学限定培养基或FCS培养的细胞的细胞壁蛋白具有复杂的图谱，有超过25条蛋白带，覆盖整个分子量范围。相比之下，从体内培养的细胞获得的消化物只有5种分子量为40 kDa或更大的主要蛋白。130 kDa和106 kDa的蛋白存在于TSB-Fe培养的细胞和体内培养的细胞的细胞膜中，但不存在于TSB或TSB+Fe培养的细胞中。体外培养的细胞表达的一种43 kDa蛋白以及体内培养的细胞表达的52 kDa和96 kDa蛋白与植入小室的猪和经导管接种旁导管接种的猪的抗血清发生反应，但不与用TSB培养的细胞免疫的猪的超免疫血清发生反应。数据表明，在体内条件下生长的表皮葡萄球菌表达的抗原与体外生长的抗原不同。
