Eosinophil penetration through cultured human airway epithelial cell layer.
We investigated the mechanisms of eosinophil penetration and mannitol permeability through a multilayer of cultured human tracheal epithelial cells. Wells of tissue culture plates were separated into the upper and the lower chambers by the cultured epithelial cell layer. 51Cr-labeled eosinophils or 3H-mannitol were put into the lower chamber. To stimulate the epithelial cells, platelet-activating factor (PAF) and/or phorbol myristate acetate (PMA) were added to the upper chamber. After 4 h of incubation, the eosinophil penetration rate was determined as a percentage of the total count added to the lower chamber. PMA significantly increased the eosinophil penetration rate in a dose-dependent manner (4.0% at 10(-5) M), compared with control (0.67%), whereas PAF itself did not. Activation of eosinophils by the addition of PAF to the lower chamber produced a significant increase in the eosinophil penetration (6.5% at 10(-6) M), which was inhibited by staurosporine. For determining the mannitol permeability, PMA, PAF, and/or supernatant from eosinophils were added to both upper and lower chambers and incubated for 30 min. PMA induced a significant increase in the mannitol permeability (175% of controls at 10(-5) M), whereas PAF itself did not alter it. Supernatant from eosinophils activated by PAF (10(-6) M) significantly increased the permeability (451% of controls), which was blocked by staurosporine. Supernatants from AA861 (a 5-lipoxygenase inhibitor)-treated or phenidon (a phospholipase A2 inhibitor)-treated eosinophils activated by PAF failed to alter the supernatant-induced increases in mannitol permeability.(ABSTRACT TRUNCATED AT 250 WORDS)
Masuda
T
T
Department of Internal and Geriatric Medicine, Tohoku University School of Medicine, Sendai, Japan.
Yamaya
M
M
Shimura
S
S
Hoshi
H
H
Sasaki
H
H
Shirato
K
K
D000470:Alkaloids / Q000494:pharmacology; D000906:Antibodies / Q000494:pharmacology; D002465:Cell Movement; D002478:Cells, Cultured; D004804:Eosinophils / Q000166:cytology*; D004847:Epithelial Cells; D004848:Epithelium / Q000187:drug effects; D006801:Humans; D018799:Intercellular Adhesion Molecule-1 / Q000276:immunology; D008353:Mannitol; D010703:Phorbol Esters / Q000494:pharmacology; D010972:Platelet Activating Factor / Q000494:pharmacology; D019311:Staurosporine; D014132:Trachea / Q000166:cytology*; D014316:Tritium
D000470:Alkaloids; D000906:Antibodies; D010703:Phorbol Esters; D010972:Platelet Activating Factor; D014316:Tritium; D018799:Intercellular Adhesion Molecule-1; D008353:Mannitol; D019311:Staurosporine