Related Experiment Video
Updated: Jul 29, 2026

Applying an Inducible Expression System to Study Interference of Bacterial Virulence Factors with Intracellular Signaling
Published on: June 25, 2015
Apoptosis observed in BALB/3T3 cells having ingested Staphylococcus aureus
1Department of Microbiology, The Jikei University School of Medicine, Tokyo, Japan. murai-miyo@spu.ac.jp
Abstract:
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.
Insights
Internalized Staphylococcus aureus can escape host cells, multiply, and trigger apoptosis in fibroblasts. This study details the intracellular mechanisms of S. aureus infection in BALB/3T3 cells.
Area of Science:
- Microbiology
- Cell Biology
- Immunology
Background:
- Staphylococcus aureus is known to be internalized by murine fibroblasts.
- The intracellular behavior of S. aureus within host cells remains incompletely understood.
Purpose of the Study:
- To investigate the intracellular events following the ingestion of Staphylococcus aureus by BALB/3T3 fibroblast cells.
- To elucidate the mechanisms by which S. aureus interacts with and affects host cells.
Main Methods:
- BALB/3T3 cells were infected with S. aureus strains (A191, A151, Cowan I).
- Cells were treated with lysostaphin to assess bacterial viability.
- DNA fragmentation was analyzed using DNA laddering and terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling (TUNEL) assay.
- Transmission electron microscopy (TEM) was used to visualize intracellular bacterial behavior and host cell morphology.
Main Results:
- S. aureus strain A191 escaped the endosome, multiplied intracellularly, and induced apoptosis in BALB/3T3 cells within 4 hours.
- Strain A151 remained largely contained within the endosome and showed limited intracellular multiplication.
- The Cowan I strain also induced DNA fragmentation, indicating apoptosis, at a slower rate.
- Apoptotic features observed included electron-dense nuclei and plasma membrane blebbing.
Conclusions:
- Internalized Staphylococcus aureus can exhibit different intracellular fates depending on the strain.
- Escape from the endosome, intracellular multiplication, and subsequent induction of apoptosis are key events in S. aureus-induced fibroblast cell death.
- These findings provide insights into the pathogenesis of S. aureus infections at the cellular level.
Related Concept Videos
Apoptosis
The Extrinsic Apoptotic Pathway
The Intrinsic Apoptotic Pathway
Phagocytosis of Apoptotic Cells
Normal cells contain receptors that prevent them from being recognized by phagocytes.
Autophagic Cell Death
Autophagy and Apoptosis
Autophagy can activate apoptosis. In normal conditions, the autophagy activating protein Beclin-1 and pro-apoptotic...
Cellular Injury V: Apoptosis and Autophagy

