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Infection and Immunity, November 2002, p. 6399-6408, Vol. 70, No. 11
0019-9567/02/$04.00+0     DOI: 10.1128/IAI.70.11.6399-6408.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Modulation of Cytosolic Ca²⁺ Concentration in Airway Epithelial Cells by Pseudomonas aeruginosa

Tobias Jacob,^1, Rebecca J. Lee,^1, Joanne N. Engel,² and Terry E. Machen^1*

Department of Molecular and Cell Biology, University of California-Berkeley, Berkeley, California 94720-3200,¹ Departments of Medicine and Microbiology and Immunology, University of California-San Francisco Medical School, San Francisco, California 94143²

Received 19 April 2002/ Returned for modification 10 June 2002/ Accepted 27 June 2002

Modulation of cytosolic (intracellular) Ca²⁺ concentration (Ca_i) may be an important host response when airway epithelial cells are exposed to Pseudomonas aeruginosa. We measured Ca_i in Calu-3 cells exposed from the apical or basolateral surface to cytotoxic and noncytotoxic strains of P. aeruginosa. Apical addition of either noncytotoxic strains or cytotoxic strains failed to affect Ca_i over a 3-h time period, nor were changes observed after basolateral addition of noncytotoxic strains. In contrast, basolateral addition of cytotoxic strains caused a slow increase in Ca_i from 100 nM to 200 to 400 nM. This increase began after 20 to 50 min and persisted for an additional 30 to 75 min, at which time the cells became nonviable. P. aeruginosa-induced increases in Ca_i were blocked by the addition of the Ca channel blocker La³⁺ to the basolateral but not to the apical chamber. Likewise, replacing the basolateral but not the apical medium with Ca-free solution prevented P. aeruginosa-mediated changes in Ca_i. With isogenic mutants of PA103, we demonstrated that the type III secretion apparatus, the type III-secreted effector ExoU, and type IV pili were necessary for increased Ca_i. We propose that translocation of ExoU through the basolateral surface of polarized airway epithelial cells via the type III secretion apparatus leads to release of Ca stored in the endoplasmic reticulum and activation of Ca channels in the basolateral membranes of epithelial cells.

Editor: J. T. Barbieri

Present address: Department of Biology, Pennsylvania State University, University Park, PA 16801.

Present address: University of New Mexico Cancer Center, Albuquerque, NM 87131.

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