Am. J. Respir. Cell Mol. Biol., Volume 23, Number 3, September, 2000 389-395

Neutrophil Transmigration across Human Airway Epithelial Monolayers
Mechanisms and Dependence on Electrical Resistance

Joseph C. Kidney and David Proud

Divisions of Pulmonary and Critical Care Medicine and Clinical Immunology, Department of Medicine, The Johns Hopkins University School of Medicine, Baltimore, Maryland

To examine neutrophil transepithelial migration in the basolateral-to-luminal direction, bronchial epithelial cells (16HBE) were grown at an air-medium interface on the lower face of permeable supports, and resistance across each membrane was recorded before measuring neutrophil transmigration over 2 h. Subconfluent monolayers (resistance < 250 ) permitted high spontaneous migration of neutrophils (7.4 ± 1%), which was further enhanced (29.7 ± 3%) in response to interleukin (IL)-8 (100 ng/ml). Confluent monolayers (250 to 700 ) showed low spontaneous migration (2 ± 0.5%) but responded markedly to IL-8 (12.4 ± 1.3%). Left in culture, 16HBE resistances continued to increase and were associated with minimal spontaneous migration (< 0.5%) or responses to IL-8. Using cells in the 250 to 700  range, neutrophil migration to IL-8 was dose-dependent and was enhanced when epithelial cells were incubated with a combination of tumor necrosis factor- and interferon-. Neutrophil migration was stimulus-specific and was reduced by preincubation of epithelial cells with a F(ab')² anti-intercellular adhesion molecule (ICAM)-1, or by preincubation of neutrophils with anti-CD18, anti-CD11a, anti-CD11b, or anti-CD11c, but not by anti-CD11d, indicating a role for ₂-integrin-ICAM-1 interaction in the migration process.

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