Neutrophil migration across a cultured epithelial monolayer elicits a biphasic resistance response representing sequential effects on transcellular and paracellular pathways

Abstract

Abstract. Migration of polymorphonuclear leukocytes across epithelia is a hallmark of many inflammatory disease states. Neutrophils traverse epithelia by migrating through the paracellular space and crossing intercellular tight junctions. We have previously shown (Nash, S., J. Stafford, and J. L. Madara. 1987. J. Clin. Invest. 80:1104-1113, that leukocyte migration across TM monolayers, a model human intestinal epithelium, results in enhanced tight junction permeability-an effect quantitated by the use of a simple, standard electrical assay of transepithelial resistance. Here we show that detailed time course studies of the transmigrationelicited decline in resistance has two components, one of which is unrelated to junctional permeability. The initial decrease in resistance, maximal 5-13 min after initiation of transmigration, occurs despite inhibition of transmigration by an antibody to the common beta subunit of neutrophil/32 integrins, and is paralleled by an increase in transepithelial short-circuit current. Chloride ion substitution and inhibitor studies indicate that the early-phase resistance decline is not attributable to an increase in tight junction permeability but is due to decreased resistance across epithelial cells resulting from chloride secretion. Since 1"84 cells are accepted models for studies of the regulation of C1-and water secretion, our results suggest that neutrophil transmigration across mucosal surfaces (for example, respiratory and intestinal tracts) may initially activate flushing of the surface by salt and water. Equally important, these studies, by providing a concrete example of sequential transcellular and paracellular effects on transepithelial resistance, highlight the fact that this widely used assay cannot simply be viewed as a direct functional probe of tight junction permeability.

Neutrophil migration across cultured intestinal epithelial monolayers is modulated by epithelial exposure to IFN-gamma in a highly polarized fashion.

“…In inflammatory diseases of organs lined by columnar epithelia, however, the migration of PMN is not complete until PMN have also migrated across the epithelial surface (Yardley and Donowitz, 1977;Yardley, 1986). While substantial progress has been made in the understanding of PMN-endothelial interactions (Arnaout et al, 1988;Springer, 1990;Larson and Springer, 1990;Osborn, 1990;Pober and Cotran, 1990;Ruoslahti, 1991), relatively limited information is available concerning PMN-epithelial interactions Nash et al, 1987Nash et al, , 1988Migliorisi et al, 1988;Parkos et al, 1991Parkos et al, , 1992. However, it is likely that such PMN-epithelial interactions may be of substantial importance.…”

“…Studies of human mucosa in such diseases suggest that PMN transepithelial migration predates focal breakdown of the epithelial surface (Yardley and Donowitz, 1977;Yardley, 1986), and that defective epithelial barrier function also predates structural discontinuities in the mucosa (Hawker et al, 1980). As outlined above, such translocation of PMN across still-intact monolayers can, by exposure of apical membranes to a neutrophilderived secretagogue (Nash et al, 1991;Madara et al, 1992), elicits fluid secretion from crypt epithelia.…”

“…However, it is likely that such PMN-epithelial interactions may be of substantial importance. For example, we have recently described a neutrophil-derived product which elicits electrogenic CI-secretion (the basis of mucosal hydration and, when upregulated in the intestine, the basis of secretory diarrhea) (Donowitz and Welsh, 1987) from human crypt intestinal epithelial cell lines and from primary isolates of mammalian crypt cells (Nash et al, 1991;Madara et al, 1992). This neutrophil-derived secretagogue primarily expresses bioactivity when presented to the apical rather than the basolateral plasma membrane of intestinal epithelial cells.…”

“…For these reasons, T~ cells are viewed as a model for haman crypt epithelium-the site at which the bulk of PMN transepithelial migration occurs in a variety of active inflammatory intestinal diseases (Yardley and Donowitz, 1977;Yardley, 1986). Our prior studies using this epithelium as a model for examination of PMN-intestinal epithelial interactions have indicated that PMN transmigration elicits a reversible decrease in transepithelial resistance due to impalement of intercellular tight junctions (Nash et al, 1987), and requires PMN ~2-integdns, specifically CDllb/CD18 (Parkos et al, , 1992. While we have shown that PMN transmigration is quantitatively more efficient in the physiological (basolateralto-apical) direction , qualitative differences in transepithelial migration, relating to the direction of transmigration, have generally not been found (Nash et al, 1987(Nash et al, , 1988Parkos et al, 1991).…”

“…We have previously modeled the event of PMN transmigration using human peripheral blood neutrophils and cultured, human~erived, intestinal epithelial monolayers derived from the cell line %4 (Nash et al, 1987;Parkos et al, , 1992Madara et al, 1993). Such monolayers are composed of columnar epithelial cells with features similar to those of natural crypt epithelia (Madara and Dharmsathaphorn, 1985;Madara et al, 1987).…”