Limits...
IL-4 and IL-13 exposure during mucociliary differentiation of bronchial epithelial cells increases antimicrobial activity and expression of antimicrobial peptides.

Zuyderduyn S, Ninaber DK, Schrumpf JA, van Sterkenburg MA, Verhoosel RM, Prins FA, van Wetering S, Rabe KF, Hiemstra PS - Respir. Res. (2011)

Bottom Line: Using sandwich ELISA we found that also hBD-2 in apical washes was increased by both IL-4 and IL-13.Apical wash obtained from IL-4- and IL-13-treated cultures displayed increased antimicrobial activity against Pseudomonas aeruginosa compared to medium-treated cultures.In addition, differentiation in the presence of Th2 cytokines resulted in increased MUC5AC production as has been shown previously.These data suggest that prolonged exposure to Th2 cytokines during mucociliary differentiation contributes to antimicrobial defence by increasing the expression and release of selected antimicrobial peptides and mucus.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Pulmonology, Leiden University Medical Center, Leiden, The Netherlands. s.zuyderduyn@lumc.nl

ABSTRACT
The airway epithelium forms a barrier against infection but also produces antimicrobial peptides (AMPs) and other inflammatory mediators to activate the immune system. It has been shown that in allergic disorders, Th2 cytokines may hamper the antimicrobial activity of the epithelium. However, the presence of Th2 cytokines also affects the composition of the epithelial layer which may alter its function. Therefore, we investigated whether exposure of human primary bronchial epithelial cells (PBEC) to Th2 cytokines during mucociliary differentiation affects expression of the human cathelicidin antimicrobial protein (hCAP18)/LL-37 and human beta defensins (hBD), and antimicrobial activity.PBEC were cultured at an air-liquid interface (ALI) for two weeks in the presence of various concentrations of IL-4 or IL-13. Changes in differentiation and in expression of various AMPs and the antimicrobial proteinase inhibitors secretory leukocyte protease inhibitor (SLPI) and elafin were investigated as well as antimicrobial activity.IL-4 and IL-13 increased mRNA expression of hCAP18/LL-37 and hBD-2. Dot blot analysis also showed an increase in hCAP18/LL-37 protein in apical washes of IL-4-treated ALI cultures, whereas Western Blot analysis showed expression of a protein of approximately 4.5 kDa in basal medium of IL-4-treated cultures. Using sandwich ELISA we found that also hBD-2 in apical washes was increased by both IL-4 and IL-13. SLPI and elafin levels were not affected by IL-4 or IL-13 at the mRNA or protein level. Apical wash obtained from IL-4- and IL-13-treated cultures displayed increased antimicrobial activity against Pseudomonas aeruginosa compared to medium-treated cultures. In addition, differentiation in the presence of Th2 cytokines resulted in increased MUC5AC production as has been shown previously.These data suggest that prolonged exposure to Th2 cytokines during mucociliary differentiation contributes to antimicrobial defence by increasing the expression and release of selected antimicrobial peptides and mucus.

Show MeSH

Related in: MedlinePlus

Two week exposure to IL-4 or IL-13 enhances antimicrobial activity. Apical wash was harvested using a 10 mM sodium phosphate buffer pH 7.4 and added to 250 colony forming units (CFU) of log phase P. aeruginosa PAO1 for 3 h. Bacteria were plated to assess number of CFU. Data represent mean CFU/ml ± SEM using apical washes obtained from six different donors.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC3113720&req=5

Figure 4: Two week exposure to IL-4 or IL-13 enhances antimicrobial activity. Apical wash was harvested using a 10 mM sodium phosphate buffer pH 7.4 and added to 250 colony forming units (CFU) of log phase P. aeruginosa PAO1 for 3 h. Bacteria were plated to assess number of CFU. Data represent mean CFU/ml ± SEM using apical washes obtained from six different donors.

Mentions: To assess whether IL-4- and IL-13-treated ALI cultures displayed antimicrobial activity, we harvested apical wash and measured antimicrobial activity. Growth of Pseudomonas aeruginosa was inhibited by apical wash obtained from six separate ALI cultures treated with IL-4 or IL-13 (Figure 4). When compared to phosphate buffer alone, medium-treated ALI cultures also exhibited some antimicrobial activity (data not shown). We did not find any antimicrobial activity in basal medium of the IL-4- and IL-13-treated cultures.


IL-4 and IL-13 exposure during mucociliary differentiation of bronchial epithelial cells increases antimicrobial activity and expression of antimicrobial peptides.

Zuyderduyn S, Ninaber DK, Schrumpf JA, van Sterkenburg MA, Verhoosel RM, Prins FA, van Wetering S, Rabe KF, Hiemstra PS - Respir. Res. (2011)

Two week exposure to IL-4 or IL-13 enhances antimicrobial activity. Apical wash was harvested using a 10 mM sodium phosphate buffer pH 7.4 and added to 250 colony forming units (CFU) of log phase P. aeruginosa PAO1 for 3 h. Bacteria were plated to assess number of CFU. Data represent mean CFU/ml ± SEM using apical washes obtained from six different donors.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3113720&req=5

Figure 4: Two week exposure to IL-4 or IL-13 enhances antimicrobial activity. Apical wash was harvested using a 10 mM sodium phosphate buffer pH 7.4 and added to 250 colony forming units (CFU) of log phase P. aeruginosa PAO1 for 3 h. Bacteria were plated to assess number of CFU. Data represent mean CFU/ml ± SEM using apical washes obtained from six different donors.
Mentions: To assess whether IL-4- and IL-13-treated ALI cultures displayed antimicrobial activity, we harvested apical wash and measured antimicrobial activity. Growth of Pseudomonas aeruginosa was inhibited by apical wash obtained from six separate ALI cultures treated with IL-4 or IL-13 (Figure 4). When compared to phosphate buffer alone, medium-treated ALI cultures also exhibited some antimicrobial activity (data not shown). We did not find any antimicrobial activity in basal medium of the IL-4- and IL-13-treated cultures.

Bottom Line: Using sandwich ELISA we found that also hBD-2 in apical washes was increased by both IL-4 and IL-13.Apical wash obtained from IL-4- and IL-13-treated cultures displayed increased antimicrobial activity against Pseudomonas aeruginosa compared to medium-treated cultures.In addition, differentiation in the presence of Th2 cytokines resulted in increased MUC5AC production as has been shown previously.These data suggest that prolonged exposure to Th2 cytokines during mucociliary differentiation contributes to antimicrobial defence by increasing the expression and release of selected antimicrobial peptides and mucus.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Pulmonology, Leiden University Medical Center, Leiden, The Netherlands. s.zuyderduyn@lumc.nl

ABSTRACT
The airway epithelium forms a barrier against infection but also produces antimicrobial peptides (AMPs) and other inflammatory mediators to activate the immune system. It has been shown that in allergic disorders, Th2 cytokines may hamper the antimicrobial activity of the epithelium. However, the presence of Th2 cytokines also affects the composition of the epithelial layer which may alter its function. Therefore, we investigated whether exposure of human primary bronchial epithelial cells (PBEC) to Th2 cytokines during mucociliary differentiation affects expression of the human cathelicidin antimicrobial protein (hCAP18)/LL-37 and human beta defensins (hBD), and antimicrobial activity.PBEC were cultured at an air-liquid interface (ALI) for two weeks in the presence of various concentrations of IL-4 or IL-13. Changes in differentiation and in expression of various AMPs and the antimicrobial proteinase inhibitors secretory leukocyte protease inhibitor (SLPI) and elafin were investigated as well as antimicrobial activity.IL-4 and IL-13 increased mRNA expression of hCAP18/LL-37 and hBD-2. Dot blot analysis also showed an increase in hCAP18/LL-37 protein in apical washes of IL-4-treated ALI cultures, whereas Western Blot analysis showed expression of a protein of approximately 4.5 kDa in basal medium of IL-4-treated cultures. Using sandwich ELISA we found that also hBD-2 in apical washes was increased by both IL-4 and IL-13. SLPI and elafin levels were not affected by IL-4 or IL-13 at the mRNA or protein level. Apical wash obtained from IL-4- and IL-13-treated cultures displayed increased antimicrobial activity against Pseudomonas aeruginosa compared to medium-treated cultures. In addition, differentiation in the presence of Th2 cytokines resulted in increased MUC5AC production as has been shown previously.These data suggest that prolonged exposure to Th2 cytokines during mucociliary differentiation contributes to antimicrobial defence by increasing the expression and release of selected antimicrobial peptides and mucus.

Show MeSH
Related in: MedlinePlus