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Am Rev Respir Dis. 1986 May;133(5):875-81.

Epithelial permeability produced by phagocytosing neutrophils in vitro.


Neutrophils are thought to increase alveolar permeability in many types of lung injury. To investigate the contribution of neutrophils to the development of permeability pulmonary edema, we have developed an in vitro cell culture system for studying alveolar epithelial permeability. Rat alveolar type II cells, cultured for 6 to 12 days on collagen-coated Millipore filters, form a morphologically and pharmacologically polarized epithelium. The filters are mounted between 2 lucite chambers, and electrical resistance (permeability to ions) and spontaneous potential difference across the monolayer are measured continually or at frequent intervals. When neutrophils and the phagocytosable particle, opsonized zymosan (but not neutrophils or opsonized zymosan alone), were added to the apical side, the potential difference and transepithelial resistance fell dramatically after 20 min, which indicates an increase in epithelial permeability. The increase in epithelial permeability was inhibited by serum alpha-1-protease inhibitor (250 micrograms/ml), methoxysuccinyl-Ala-Ala-Pro-Val-chloromethyl ketone (0.02 mM) (an elastase inhibitor), catalase (2,500 units/ml), and superoxide dismutase (330 units/ml). In experiments with a lower concentration of phagocytosing neutrophils, a slower rate of decrease in resistance occurred, and in 3 of 13 studies, there was a definite recovery of the resistance to initial values. This study demonstrated that phagocytosing but not resting neutrophils increase the permeability of the epithelial monolayers to ions and suggests that the increased permeability in this system is mediated in part by both neutral protease(s) and oxygen radicals.

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