title: Modulation of neutrophil and mononuclear cell adherence to bronchial epithelial cells.
abstract: Neutrophils and mononuclear cells have been associated with the lower respiratory tract inflammation observed in both acute and chronic bronchitis. In order to transit into and remain within the airways, neutrophils and mononuclear cells would likely need to adhere to bronchial epithelium. To test this hypothesis, bovine bronchial epithelial cells (BBECs) were isolated and cultured on a round coverslip. After 7 to 10 days, 51Cr-labeled neutrophils and mononuclear cells were evaluated for their capacity to adhere to the BBEC monolayer. Both neutrophils and mononuclear cells readily bound to the BBEC monolayer (10.8 +/- 1.2% bound neutrophils; 40.5 +/- 2.8% bound mononuclear cells). Stimulation of the neutrophils and mononuclear cells with phorbol 12-myristate 13-acetate (PMA) increased the adherence (45.8 +/- 10.6% bound neutrophils, P less than 0.01 compared with unstimulated cells; 58.7 +/- 6.2% bound mononuclear cells, P less than 0.01 compared with unstimulated cells). Importantly, stimulating the BBEC monolayer with PMA, bacterial lipopolysaccharide, or a cigarette smoke extract for 4 to 72 h also increased the adherence of both cell types (P less than 0.01, all comparisons at 24 h). The adherence was not decreased by exposure of either the BBEC monolayer, the neutrophils, or the mononuclear cells to cycloheximide or to the anti-CD11/CD18 monoclonal antibody 60.3 (P greater than 0.05). However, exposure of the BBEC monolayer to trypsin before addition of the neutrophils significantly decreased adherence (P less than 0.05). Because neutrophils and mononuclear cells are thought to mediate cell cytotoxicity by adhering to the target cells, BBECs were labeled with 51Cr, and 51Cr release was measured as an index of cytotoxicity. There was a modest increase in 51Cr release by the addition of unstimulated neutrophils and mononuclear cells, and culturing the BBEC monolayer with PMA before the addition of the neutrophils or mononuclear cells resulted in a further modest enhancement of 51Cr release (P less than 0.05). Similar results were obtained using lactate dehydrogenase release as a measure of cytotoxicity. These results demonstrate that inflammatory cells can adhere to BBECs and may be capable of mediating cytotoxicity and adherence and cytotoxicity can be increased by stimulating BBECs.
authors.affiliation: Research Service, Omaha Veterans Affairs Medical Center, Nebraska.
authors.forename: R A
authors.initials: RA
authors.lastname: Robbins
authors.forename: S
authors.initials: S
authors.lastname: Koyama
authors.forename: J R
authors.initials: JR
authors.lastname: Spurzem
authors.forename: K A
authors.initials: KA
authors.lastname: Rickard
authors.forename: K J
authors.initials: KJ
authors.lastname: Nelson
authors.forename: G L
authors.initials: GL
authors.lastname: Gossman
authors.forename: G M
authors.initials: GM
authors.lastname: Thiele
authors.forename: S I
authors.initials: SI
authors.lastname: Rennard
mesh_terms: D000818:Animals; D001980:Bronchi / Q000166:cytology* / Q000276:immunology / Q000648:ultrastructure; D002417:Cattle; D002448:Cell Adhesion; D002478:Cells, Cultured; D003513:Cycloheximide / Q000494:pharmacology; D003602:Cytotoxicity, Immunologic; D004847:Epithelial Cells; D004848:Epithelium / Q000276:immunology / Q000648:ultrastructure; D006801:Humans; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D007963:Leukocytes, Mononuclear / Q000166:cytology* / Q000187:drug effects / Q000276:immunology / Q000648:ultrastructure; D008855:Microscopy, Electron, Scanning; D009504:Neutrophils / Q000166:cytology* / Q000187:drug effects / Q000276:immunology / Q000648:ultrastructure; D012397:Rosette Formation; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D014357:Trypsin
chemical_list: D003513:Cycloheximide; D007770:L-Lactate Dehydrogenase; D014357:Trypsin; D013755:Tetradecanoylphorbol Acetate