Effects of topical glucocorticoids on in vitro lactoferrin glandular secretion: Comparison between human upper and lower airways Jordi Roca-Ferrer, MSa,

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Effects of topical glucocorticoids on in vitro lactoferrin glandular secretion: Comparison between human upper and lower airways Jordi Roca-Ferrer, MSa, Joaquim Mullol, MDa, c, María Pérez, BSa, Antoni Xaubet, MDa, b, Laureano Molins, MDd, Josep de Haro, MDe, James Shelhamer, MDf, César Picado, MDa, b Journal of Allergy and Clinical Immunology Volume 106, Issue 6, Pages 1053-1062 (December 2000) DOI: 10.1067/mai.2000.110476 Copyright © 2000 Mosby, Inc. Terms and Conditions

Fig. 1 Lactoferrin location in nasal and bronchial mucosa. Immunoreactive material to lactoferrin (arrows) is located in serous cells in both nasal (A) and bronchial (B) submucosal glands (s, serous cells; m, mucous cells). Hematoxylin-and-eosin staining shows (C) the structure of human nasal mucosa (ep, epithelium; bm, basal membrane; sm, submucosa) and (D) bronchial glands (g) . There was a total absence of CD15+ cells in both nasal (E) and bronchial (F) explants after 48 hours of incubation with culture media. Journal of Allergy and Clinical Immunology 2000 106, 1053-1062DOI: (10.1067/mai.2000.110476) Copyright © 2000 Mosby, Inc. Terms and Conditions

Fig. 2 Spontaneous lactoferrin release from human airway mucosa in culture. Once placed in culture and after a 48-hour stabilization period (days –2 to –1), the concentration of lactoferrin in supernatants from both nasal (open triangles , n = 12) and bronchial (open circles , n = 6) explants progressively decreased during the 6 days in culture. ANOVA with Dunnett t test comparisons was used for analysis (*P < .05 and **P < .01 compared with lactoferrin secretion on day 0). Journal of Allergy and Clinical Immunology 2000 106, 1053-1062DOI: (10.1067/mai.2000.110476) Copyright © 2000 Mosby, Inc. Terms and Conditions

Fig. 3 Effects of topical corticosterids on spontaneous lactoferrin release from nasal and bronchial mucosa. A, After culture stabilization, explants were incubated with selected corticosteroids for 48 hours (2 days), and culture supernatants were collected every 24 hours (arrows) . B, Both budesonide (filled circles) and beclomethasone dipropionate (open circles) inhibited the spontaneous lactoferrin secretion from nasal explants. The effect of budesonide was significant from day 2 to day 5, with a maximum effect at day 5. The effect of beclomethasone dipropionate was significant on days 4 and 5, when its maximum effect was reached. C, Both budesonide (filled squares) and beclomethasone dipropionate (open squares) also showed an inhibitory effect on the baseline secretion of lactoferrin from bronchial explants. The effect of budesonide was significant from day 3 to day 5, reaching its maximum effect on day 3. Beclomethasone dipropionate caused a significant inhibition of lactoferrin secretion from day 1 to day 3, also reaching its maximum effect on day 3. ANOVA with Dunnett t test comparisons was used for analysis (*P < .05 and **P < .01 compared with media-treated explants). Journal of Allergy and Clinical Immunology 2000 106, 1053-1062DOI: (10.1067/mai.2000.110476) Copyright © 2000 Mosby, Inc. Terms and Conditions

Fig. 4 Effect of topical corticosteroids on cholinergic-induced lactoferrin release from nasal and bronchial mucosa. A, After stabilization, explants were incubated with selected corticosteroids for 48 hours (2 days) and, after a washout period (24 hours), methacholine (MCh, 10–6 mol/L) was added for 2 hours. B, In nasal mucosa explant incubation with budesonide (filled bars , n = 12) or beclomethasone dipropionate (hatched bars , n = 7) from 10–10 to 10–6 mol/L caused a dose-related inhibition of lactoferrin secretion induced by methacholine at 10–6 mol/L (budesonide: IC50 of 181 nmol/L; beclomethasone dipropionate: IC50 of 136 nmol/L). C, In bronchial mucosa budesonide (filled bars , n = 6) from 10–12 to 10–6 mol/L also caused a dose-related inhibition (IC50 of 15 nmol/L) of lactoferrin secretion induced by methacholine at 10–6 mol/L (open bar) . The Wilcoxon signed-rank test was used for analysis (*P < .05 and **P < .01). Journal of Allergy and Clinical Immunology 2000 106, 1053-1062DOI: (10.1067/mai.2000.110476) Copyright © 2000 Mosby, Inc. Terms and Conditions