Wallengren M L Lundin, Hamberg K, Ericson D
Department of Cariology, Faculty of Odontology, Malmö University, Sweden.
Oral Microbiol Immunol. 2004 Jun;19(3):188-95. doi: 10.1111/j.0902-0055.2004.00141.x.
In the immunoblot technique, using whole bacteria cell extracts as antigens, both intra- and extracellular antigens are detected, which gives a large number of immunoglobulin A (IgA) reactions (immunoblot bands) when incubated with saliva. It is important to distinguish which immunoblot bands represent bacterial cell-surface antigens, since these antigens could be involved in adhesion mechanisms and be available for blocking in vivo.
Bacterial extracts of Streptococcus mutans, Streptococcus sobrinus, Streptococcus parasanguis and the streptococcal antigen I/II were separated using SDS-PAGE. The antigens were detected with saliva in Western blot. Untreated saliva and saliva in which cell-surface reactive IgA had been absorbed with whole bacteria cells were analyzed.
Approximately half the number of the bands were absent for saliva absorbed with homologous cells, compared to untreated saliva. The absorption pattern was almost identical for S. mutans and S. sobrinus but not for S. parasanguis. Salivary IgA reactive against streptococcal antigen I/II was absorbed by S. mutans cells, to a lesser extent by S. sobrinus cells, and not at all by S. parasanguis cells.
It is likely that the bands that were absent after absorption represented cell-surface antigens. For S. mutans and S. sobrinus, these bands were probably the streptococcal antigen I/II.
在免疫印迹技术中,使用全细菌细胞提取物作为抗原,细胞内和细胞外抗原均可被检测到,当与唾液孵育时会产生大量免疫球蛋白A(IgA)反应(免疫印迹条带)。区分哪些免疫印迹条带代表细菌细胞表面抗原很重要,因为这些抗原可能参与黏附机制并且可用于体内阻断。
使用十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)分离变形链球菌、远缘链球菌、血链球菌的细菌提取物以及链球菌抗原I/II。在蛋白质印迹法中用唾液检测抗原。分析未处理的唾液以及细胞表面反应性IgA已被全细菌细胞吸收的唾液。
与未处理的唾液相比,用同源细胞吸收后的唾液中条带数量减少了约一半。变形链球菌和远缘链球菌的吸收模式几乎相同,但血链球菌不同。针对链球菌抗原I/II的唾液IgA被变形链球菌细胞吸收,被远缘链球菌细胞部分吸收,而血链球菌细胞则完全不吸收。
吸收后消失的条带可能代表细胞表面抗原。对于变形链球菌和远缘链球菌,这些条带可能是链球菌抗原I/II。