Shaw J H, Falkow S
Department of Medical Microbiology, Stanford University School of Medicine, California 94305.
Infect Immun. 1988 Jun;56(6):1625-32. doi: 10.1128/iai.56.6.1625-1632.1988.
A tissue culture model has been developed for studying the ability of Neisseria gonorrhoeae to invade eucaryotic cells. The cell line HecIB, a human adenocarcinoma endometrial cell line, was found to support gonococcal invasion. The bactericidal antibiotic gentamicin was used to kill those bacteria that had not entered the HecIB cells, allowing us to quantitate internalized bacteria. Kinetic studies showed an increase in the titer of gentamicin-protected gonococci at 4 h postinfection followed by a decrease; a second increase occurred after 6 h. The state of piliation did not affect the degree of invasion when the bacteria were spun down onto the monolayer. Gonococcal invasion was inhibited when the HecIB cells were preincubated with cytochalasin D before bacterial infection. N. lactamica was used as a negative control. No internalized N. lactamica cells were observed by electron microscopy. Electron microscopy documented the intracellular location of the gonococci in HecIB cells and the eventual destruction of the invaded HecIB cells. After 24 h, clusters of gonococci encased in a matrix of cell debris were observed.
已建立一种组织培养模型，用于研究淋病奈瑟菌侵入真核细胞的能力。发现人子宫内膜腺癌细胞系HecIB能支持淋球菌的侵入。使用杀菌抗生素庆大霉素杀死未进入HecIB细胞的细菌，从而使我们能够对内化细菌进行定量。动力学研究表明，感染后4小时，受庆大霉素保护的淋球菌滴度增加，随后下降；6小时后出现第二次增加。当将细菌离心到单层细胞上时，菌毛状态不影响侵入程度。在细菌感染前，用细胞松弛素D预孵育HecIB细胞可抑制淋球菌的侵入。乳酸奈瑟菌用作阴性对照。通过电子显微镜未观察到内化的乳酸奈瑟菌细胞。电子显微镜记录了淋球菌在HecIB细胞内的位置以及被侵入的HecIB细胞最终的破坏情况。24小时后，观察到包裹在细胞碎片基质中的淋球菌簇。
