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Infection and Immunity, September 2000, p. 5385-5392, Vol. 68, No. 9
0019-9567/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Staphylococcus aureus RN6390 Replicates and Induces Apoptosis in a Pulmonary Epithelial Cell Line

Barbara C. Kahl,1 Mark Goulian,2 Willem van Wamel,3 Mathias Herrmann,4 Sanford M. Simon,2 Gilla Kaplan,5 Georg Peters,4 and Ambrose L. Cheung3,*

Laboratory of Bacterial Pathogenesis and Immunology,1 Laboratory of Cellular Biophysics,2 and Laboratory of Cellular Physiology and Immunology,5 Rockfeller University, New York, New York 10021; Department of Microbiology, Dartmouth Medical School, Hanover, New Hampshire 032553; and Medical Microbiology, Westfälische Wilhelms Universität Münster, D-48149 Münster, Germany4

Received 20 March 2000/Returned for modification 10 May 2000/Accepted 26 May 2000

Staphylococcus aureus frequently colonizes the airways of patients with compromised airway defenses (e.g., cystic fibrosis [CF] patients) for extended periods. Persistent and relapsing infections may be related to live S. aureus bacteria actively residing inside epithelial cells. In this study, we infected a respiratory epithelial cell line, which was derived from a CF patient, with S. aureus RN6390. Internalization of S. aureus was found to be time and dose dependent and could be blocked by cytochalasin D. Transmission electron microscopy revealed that internalized bacteria resided within endocytic vacuoles without any evidence of lysosomal fusion in a 24-h period. The results of internalization experiments and time-lapse fluorescence microscopy of epithelial cells infected with green fluorescent S. aureus indicate that, after an initial lag period of 7 to 9 h, intracellular bacteria began to replicate, with three to five divisions in a 24-h period, leading to apoptosis of infected cells. Induction of apoptosis required bacterial internalization and is associated with intracellular replication. The slow and gradual replication of S. aureus inside epithelial cells hints at the role of host factors or signals in bacterial growth and further suggests possible cross talk between host cells and S. aureus.

* Corresponding author. Mailing address: Department of Microbiology, Dartmouth Medical School, Hanover, NH 03255. Phone: (603) 650-1340. Fax: (603) 650-1318. E-mail: Ambrose.Cheung{at}Dartmouth.edu.

Infection and Immunity, September 2000, p. 5385-5392, Vol. 68, No. 9
0019-9567/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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