Wise K S, Watson R K
Infect Immun. 1983 Sep;41(3):1332-9. doi: 10.1128/iai.41.3.1332-1339.1983.
Four antigens of Mycoplasma hyorhinis GDL were defined by murine monoclonal antibodies. Components of broth-grown mycoplasmas were separated under reducing conditions by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and subsequent protein blots were stained with individual antibodies. Each antibody reacted with a distinct component with relative molecular weights of 120,000, 73,000, 51,000, and 38,000, respectively (termed p120, p73, p51, and p38). Trypsin treatment of protein blots specifically abrogated binding of antibodies, suggesting that the epitopes recognized were associated with proteins. By using indirect immunofluorescence and immunoferritin techniques, mycoplasmas colonizing the surface of chronically infected BW5147 murine T-lymphoblastoid cells were selectively stained with antibody to p120, indicating the localization of the corresponding epitope at the mycoplasma surface. Protein blots of mycoplasmas derived from BW5147 cell cultures were stained with antibody to p120, revealing a component identical to that observed with broth-grown organisms. These results establish the identity of a surface protein antigen of M. hyorhinis GDL expressed at the surface of organisms during their colonization of host cells.
鼠单克隆抗体鉴定出了猪鼻支原体GDL的四种抗原。在还原条件下，通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳分离肉汤培养的支原体成分，随后用各单克隆抗体对蛋白质印迹进行染色。每种抗体分别与相对分子质量为120,000、73,000、51,000和38,000的不同成分发生反应（分别称为p120、p73、p51和p38）。对蛋白质印迹进行胰蛋白酶处理可特异性消除抗体结合，这表明所识别的表位与蛋白质相关。通过间接免疫荧光和免疫铁蛋白技术，用抗p120抗体对慢性感染的BW5147鼠T淋巴细胞样细胞表面定植的支原体进行选择性染色，表明相应表位定位于支原体表面。用抗p120抗体对源自BW5147细胞培养物的支原体蛋白质印迹进行染色，显示出与肉汤培养的生物体中观察到的成分相同的一种成分。这些结果确定了猪鼻支原体GDL的一种表面蛋白抗原的特性，该抗原在生物体定植于宿主细胞的过程中在其表面表达。
