Sakai D K
Infect Immun. 1987 Mar;55(3):704-10. doi: 10.1128/iai.55.3.704-710.1987.
The adhesion of Aeromonas salmonicida, the pathogenic bacterium of fish furunculosis in salmon and trout, to the surface of host tissue cells was investigated with two fish tissue culture cell lines (RTG-2 cells from rainbow trout, Salmo gairdneri, and CHSE-214 cells from chinook salmon, Oncorhynchus tshawytscha) and four A. salmonicida strains. Bacterial cells of pathogenic strains were highly adhesive to RTG-2 and CHSE-214 cells and were negatively charged in the net electrostatic charges, as determined by electrophoresis on filter paper strips at pH 7, whereas bacterial cells of nonpathogenic strains were nonadhesive and positively charged. The electrophoresis of RTG-2 and CHSE-214 cells with balanced salt solution (BSS), phosphate-buffered saline, or fish serum diluted with BSS (pH 7) was carried out with an appropriate electrophoretic apparatus that was devised for this study. After electrophoresis with 20 mA of direct current for 15 min at pH 7, the electrophoretic dispositions of these tissue culture cells were determined by the mode of frequency of occurrence of these cells in the partitioned chambers of the device. RTG-2 and CHSE-214 cells with BSS and fish serum were attracted from the central chamber (to which each cell sample was added) to the cathode chambers, but no attraction was detected when these cells were used with phosphate-buffered saline. Noradrenaline- and phosphoenolpyruvate-pretreated RTG-2 cells migrated more to the cathode chambers, whereas succinate- and valine-pretreated RTG-2 cells moved to the anode chambers. These movements to the cathode and anode were alleviated by the use of RTG-2 cells preincubated with pathogenic and nonpathogenic bacterial cells, respectively. The adhesion of the pathogenic bacteria to RTG-2 cells was enhanced by the use of RTG-2 cells pretreated with noradrenaline and phosphoenolpyruvate, whereas the nonpathogenic bacteria were adherent to RTG-2 cells pretreated with succinate and valine. These findings indicate that the adhesion of A. salmonicida strains to host tissue cells is closely associated with mutually converse net electrostatic charges.
利用两种鱼类组织培养细胞系(虹鳟鱼(Salmo gairdneri)的RTG - 2细胞和奇努克鲑鱼(Oncorhynchus tshawytscha)的CHSE - 214细胞)以及四株杀鲑气单胞菌,研究了鲑鱼和鳟鱼疖疮病的病原菌——杀鲑气单胞菌对宿主组织细胞表面的黏附情况。通过在pH 7的滤纸条上进行电泳测定,致病菌株的细菌细胞对RTG - 2和CHSE - 214细胞具有高度黏附性,且净静电荷为负,而非致病菌株的细菌细胞无黏附性且带正电荷。使用为本研究设计的合适电泳装置,对RTG - 2和CHSE - 214细胞在平衡盐溶液(BSS)、磷酸盐缓冲盐水或用BSS稀释的鱼血清(pH 7)中进行电泳。在pH 7下以20 mA直流电电泳15分钟后,通过这些细胞在装置分隔腔室中的出现频率模式来确定这些组织培养细胞的电泳分布。含有BSS和鱼血清的RTG - 2和CHSE - 214细胞从中央腔室(每个细胞样品加入此处)被吸引到阴极腔室,但当这些细胞与磷酸盐缓冲盐水一起使用时未检测到吸引现象。用去甲肾上腺素和磷酸烯醇丙酮酸预处理的RTG - 2细胞向阴极腔室迁移得更多,而用琥珀酸和缬氨酸预处理的RTG - 2细胞则向阳极腔室移动。分别用致病和非致病细菌细胞预孵育的RTG - 2细胞可减轻这些向阴极和阳极的移动。用去甲肾上腺素和磷酸烯醇丙酮酸预处理的RTG - 2细胞可增强致病细菌对RTG - 2细胞的黏附,而用琥珀酸和缬氨酸预处理的RTG - 2细胞则使非致病细菌黏附。这些发现表明,杀鲑气单胞菌菌株对宿主组织细胞的黏附与相互相反的净静电荷密切相关。