Apoptosis observed in BALB/3T3 cells having ingested Staphylococcus aureus.
Staphylococcus aureus was previously shown to be internalized by murine fibroblast. We examined the intracellular events of S. aureus ingested by BALB/3T3 cells. After uptake of strains A191 and A151, isolates from atopic lesion, and a laboratory strain, Cowan I, for 1 hr, BALB/3T3 cells were incubated with 1.25 microg/ml lysostaphin. Laddering of the DNA in multiples of approximately 180 bp occurred within 4 hr following bacterial addition in BALB/3T3 cells infected with A191 and within 18 hr in BALB/3T3 cells infected with A151: histochemical staining by the terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling method revealed that the rate of the fragmentation of nucleic DNA in Cowan I-infected BALB/3T3 cells at 21 hr following bacterial addition was 0.52 +/- 0.25%, significantly higher than that in the control cells. Transmission electron micrographs of BALB/3T3 cells at 4 hr following A191 addition showed that the apoptotic features, including electron-dense nucleus and plasma membrane blebbing, occurred in some cells in which many staphylococci escaped the endosome and went on to cell division. At the same time, A151 organisms enclosed with endosome membrane were static in the intact BALB/3T3 cells. The significant increase of A191 was confirmed by counting intracellular live bacteria during 2- to 6-hr incubation. These results suggest that internalized S. aureus escapes the endosome, multiplies and induces apoptosis in the fibroblast cell.
Murai
M
M
Department of Microbiology, The Jikei University School of Medicine, Tokyo, Japan. murai-miyo@spu.ac.jp
Sakurada
J
J
Seki
K
K
Shinji
H
H
Hirota
Y
Y
Masuda
S
S
D016475:3T3 Cells; D000818:Animals; D017209:Apoptosis; D053938:DNA Fragmentation; D007424:Intracellular Fluid; D051379:Mice; D008807:Mice, Inbred BALB C; D008854:Microscopy, Electron; D013211:Staphylococcus aureus / Q000502:physiology*