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Am. J. Respir. Cell Mol. Biol., Volume 23, Number 3, September, 2000 304-312

Pseudomonas aeruginosa Induction of Apoptosis in Respiratory Epithelial Cells
Analysis of the Effects of Cystic Fibrosis Transmembrane Conductance Regulator Dysfunction and Bacterial Virulence Factors

Sujatha Rajan, Grace Cacalano, Ruth Bryan, Adam J. Ratner, Claudia U. Sontich, Anna van Heerckeren, Pamela Davis, and Alice Prince

Department of Pediatric Infectious Diseases, College of Physicians & Surgeons, Columbia University, New York, New York; and Department of Pediatrics, Case Western Reserve University, Cleveland, Ohio

Airway epithelial cells can respond to infection by activating several signaling pathways. We examined the induction of apoptosis in response to Pseudomonas aeruginosa PAO1 in normal cells and several cystic fibrosis (CF) and corrected cell lines. Epithelial cells in monolayers with tight junctions, confirmed by apical ZO-1 staining demonstrated by confocal microscopy, were entirely resistant to PAO1-induced apoptosis. In contrast, cell lines such as 9HTEo- cells that do not form tight junctions were susceptible, with 50% of the population apoptotic after 6 h of exposure to PAO1. CF transmembrane conductance regulator (CFTR) dysfunction caused by different mechanisms (trafficking mutations, overexpression of the regulatory domain or antisense constructs) did not alter rates of apoptosis, nor were differences apparent in terminal deoxyribonucleotidyl transferase-mediated deoxyuridine triphosphate-biotin nick-end labeling detection of apoptotic airway cells from PAO1 infected cftr -/- or control mice. Bacterial expression of specific adhesins, complete lipopolysaccharide, and a functional type III secretion system were all necessary to evoke apoptosis even in susceptible epithelial cells. Unlike other mucosal surfaces, the airway epithelium is highly resistant to apoptosis, and this response is activated only when the appropriate epithelial conditions are present as well as fully virulent P. aeruginosa capable of coordinately expressing both adhesins and cytotoxins.




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