Boissinot M, Ramsay D, Barthe C, Joly J R
Département de Microbiologie, Faculté de Médecine, Université Laval, Québec, Canada.
Can J Microbiol. 1987 Jul;33(7):607-13. doi: 10.1139/m87-106.
The outer membrane proteins of Legionella pneumophila serogroups 1 to 8 were prepared from broken cells by selective solubilization using sodium lauryl sarcosinate. The isolated proteins were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and transferred to nitrocellulose sheets. Rabbit antisera against each of the eight serogroups of L. pneumophila were obtained by immunizing each animal with live bacteria. The transferred proteins were revealed using these antisera and peroxidase-labeled swine anti-rabbit immunoglobulins. Antigenic determinants common to all eight serogroups were found in at least three outer membrane antigens (19, 29, and 45 kilodaltons (kDa)). However, cross-absorption experiments revealed that these three antigens were immunologically related, but not identical among serogroups. The antigenic relationships observed with two of these three antigens correlated well with cross-reactions observed in immunofluorescence. When a monoclonal antibody directed against L. pneumophila serogroup 1 lipopolysaccharide was used to reveal a blot of serogroup 1 outer membrane antigens, the 29- and 45-kDa bands appeared. This demonstrates a strong association between lipopolysaccharide and outer membrane proteins.
嗜肺军团菌血清型1至8的外膜蛋白是通过使用月桂醇肌氨酸钠选择性溶解破碎细胞而制备的。分离得到的蛋白质通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳进行分离，并转移至硝酸纤维素膜上。通过用活细菌免疫每只动物，获得了针对嗜肺军团菌八个血清型中每个血清型的兔抗血清。使用这些抗血清和过氧化物酶标记的猪抗兔免疫球蛋白来显示转移的蛋白质。在至少三种外膜抗原（19、29和45千道尔顿（kDa））中发现了所有八个血清型共有的抗原决定簇。然而，交叉吸收实验表明，这三种抗原在免疫学上相关，但在血清型之间并不相同。观察到的这三种抗原中的两种之间的抗原关系与免疫荧光中观察到的交叉反应密切相关。当使用针对嗜肺军团菌血清型1脂多糖的单克隆抗体来显示血清型1外膜抗原的印迹时，出现了29 kDa和45 kDa的条带。这证明了脂多糖与外膜蛋白之间存在强关联。
