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Am J Physiol Lung Cell Mol Physiol 278: L528-L535, 2000;
1040-0605/00 $5.00
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Vol. 278, Issue 3, L528-L535, March 2000

Mesothelial cell apoptosis is confirmed in vivo by morphological change in cytokeratin distribution

E. Marchi, W. Liu, and V. C. Broaddus

Lung Biology Center, San Francisco General Hospital, San Francisco, California 94143-0854

Apoptosis of mesothelial cells has been demonstrated in vitro but not in vivo. To identify apoptotic pleural cells as mesothelial, we used cytokeratin as a marker and found a striking spheroid, aggregated appearance of cytokeratin in apparently apoptotic mesothelial cells. In in vitro studies, we found that the aggregated cytokeratin pattern correlated with apoptosis in primary mesothelial cells from mice, rabbits, and humans and was not seen with necrosis. In in vivo studies in mice, we then used this cytokeratin pattern to identify and quantitate apoptotic mesothelial cells. Apoptotic mesothelial cells were best harvested by pleural lavage, indicating that they were loosely adherent or nonadherent. Instillation of RPMI 1640 medium or wollastonite for 24 h induced apoptosis in 0.1 ± 0.1 (SE) and 1.0 ± 0.7%, respectively, of all mesothelial cells recovered, whereas instillation of known apoptotic stimuli, crocidolite asbestos (25 µg) for 24 h or actinomycin D plus murine tumor necrosis factor-alpha for 12 h, induced apoptosis in 5.1 ± 0.5 and 22.4 ± 4.5%, respectively (significantly greater than in control experiments, P < 0.05). By analysis of cytokeratin staining, mesothelial cell apoptosis has been confirmed in vivo.

crocidolite asbestos; wollastonite; keratin 18; pleura; intermediate filament


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