Merkel G J, Phelps C L
Department of Microbiology and Immunology, Indiana University School of Medicine, Fort Wayne 46805.
Infect Immun. 1988 Apr;56(4):792-801. doi: 10.1128/iai.56.4.792-801.1988.
The interaction of Candida albicans clinical isolates with primary and established fibroblast cultures was studied. The intent was to determine whether yeast adherence and invasion of nonendothelial cell monolayer cultures could be quantitated reproducibly and whether this system could be used for future studies on yeast pathogenesis. Our results demonstrated that specific interactions between the yeast cells and fibroblasts only occurred at 37 degrees C and correlated with the germination process. Fluorescent-antibody staining indicated that invasion or tight associations between the germinating yeast cells and mammalian cells occurred after less than 3 h of incubation. Yeast adherence was estimated radiometrically and trypsin-resistant interaction with individual mammalian cells (infection) was measured microscopically after inoculated monolayer cells were detached with trypsin. We demonstrated that both types of association were time dependent at 37 degrees C; neither was affected by the concentration of glucose used to grow the yeast cells. Primary and established fibroblast cell lines were equally susceptible to infection, but primary cells appeared to have more yeast-binding sites. Fibroblasts maintained in confluent culture for an extended period of time also appeared to have more binding sites, and while not quantitatively more susceptible to infection, the older cells were more susceptible to infection-related cell death. An established kidney epithelial cell line (MDCK) was not susceptible to either type of yeast interaction, indicating that the yeast-fibroblast associations were specific.
研究了白色念珠菌临床分离株与原代及已建立的成纤维细胞培养物之间的相互作用。目的是确定酵母对非内皮细胞单层培养物的黏附和侵袭是否能够被可重复地定量,以及该系统是否可用于未来关于酵母致病机制的研究。我们的结果表明,酵母细胞与成纤维细胞之间的特异性相互作用仅在37℃时发生,且与萌发过程相关。荧光抗体染色表明,在孵育不到3小时后,萌发的酵母细胞与哺乳动物细胞之间发生了侵袭或紧密关联。通过放射性测量估计酵母黏附情况,并在用胰蛋白酶使接种的单层细胞脱离后,通过显微镜测量与单个哺乳动物细胞的抗胰蛋白酶相互作用(感染)。我们证明,在37℃时,这两种关联类型均与时间相关;两者均不受用于培养酵母细胞的葡萄糖浓度的影响。原代及已建立的成纤维细胞系对感染同样敏感,但原代细胞似乎具有更多的酵母结合位点。长时间维持在汇合培养状态的成纤维细胞似乎也具有更多的结合位点,并且虽然在数量上对感染的敏感性没有更高,但较老的细胞对与感染相关的细胞死亡更敏感。一种已建立的肾上皮细胞系(MDCK)对酵母的两种相互作用类型均不敏感,这表明酵母与成纤维细胞的关联是特异性的。