Kihlström E
Infect Immun. 1977 Aug;17(2):290-5. doi: 10.1128/iai.17.2.290-295.1977.
After interaction with HeLa cells cultured in vitro, the fraction of adhering extracellular and that of internalized smooth Salmonella typhimurium 395 MS and rough 395 MR10 have been determined by two different techniques. (i) By using the indirect fluorescent-antibody technique on unfixed and acetone-fixed HeLa cell preparations, intracellular bacteria were considered to become stained only after acetone fixation. (ii) Based on the assumption that gentamicin affects only extracellular bacteria, disintegration of the infected HeLa cells and viable count allowed the determination of internalized bacteria. Both techniques showed that MS as well as MR10 bacteria gained intracellular access, the fraction of MR10 cells doing so being much greater. The net increase in the intracellular bacterial population was small within 3 h of incubation.
在与体外培养的HeLa细胞相互作用后,已通过两种不同技术测定了粘附的细胞外鼠伤寒沙门氏菌光滑型395 MS和粗糙型395 MR10内化部分的比例。(i)通过对未固定和经丙酮固定的HeLa细胞制剂使用间接荧光抗体技术,细胞内细菌被认为仅在丙酮固定后才会被染色。(ii)基于庆大霉素仅影响细胞外细菌的假设,对感染的HeLa细胞进行裂解并进行活菌计数,从而确定内化细菌的数量。两种技术均显示MS和MR10细菌都能进入细胞内,其中MR10细胞进入细胞内的比例要大得多。在孵育3小时内,细胞内细菌群体的净增加量很小。