TY - JOUR
T1 - EGR-1 and DUSP-1 are important negative regulators of pro-allergic responses in airway epithelium
AU - Golebski, Korneliusz
AU - van Egmond, Danielle
AU - de Groot, Esther J.
AU - Roschmann, Kristina I. L.
AU - Fokkens, Wytske J.
AU - van Drunen, Cornelis M.
PY - 2015
Y1 - 2015
N2 - Background: Primary nasal epithelium of house dust mite allergic individuals is in a permanently activated inflammatory transcriptional state. Objective: To investigate whether a deregulated expression of EGR-1 and/or DUSP-1, two potential negative regulators of pro-inflammatory responses, could contribute to the activation of the inflammatory state. Methods: We silenced the expression of EGR-1 or DUSP-1 in the airway epithelial cell line NCI-H292. The cell lines were stimulated in a 24-h time course with the house dust mite allergen or poly(I:C). RNA expression profiles of cytokines were established using q-PCR and protein levels were determined in supernatants with ELISA. Results: The shRNA-mediated gene silencing reduced expression levels of EGR-1 by 92% (p <0.0001) and of DUSP-1 by 76% (p <0.0001). Both mutant cells lines showed an increased and prolonged response to the HDM allergen. The mRNA induction of IL-6 was 4.6 fold (p = 0.02) and 2.4 fold higher (p = 0.01) in the EGR-1 and DUSP-1 knock-down, respectively when compared to the induced levels in the control cell line. For IL-8, the induction levels were 4.6 fold (p = 0.01) and 13.0 (p = 0.001) fold higher. The outcome was largely similar, yet not identical at the secreted protein levels. Furthermore, steroids were able to suppress the poly(I:C) induced cytokine levels by 70-95%. Conclusions: Deregulation of EGR-1 and/or DUSP-1 in nasal epithelium could be responsible for the prolonged activated transcriptional state observed in vivo in allergic disease. This could have clinical consequences as cytokine levels after the steroid treatment in EGR-1 or DUSP-1 knock-down remained higher than in the control cell line. (C) 2015 Elsevier Ltd. All rights reserved
AB - Background: Primary nasal epithelium of house dust mite allergic individuals is in a permanently activated inflammatory transcriptional state. Objective: To investigate whether a deregulated expression of EGR-1 and/or DUSP-1, two potential negative regulators of pro-inflammatory responses, could contribute to the activation of the inflammatory state. Methods: We silenced the expression of EGR-1 or DUSP-1 in the airway epithelial cell line NCI-H292. The cell lines were stimulated in a 24-h time course with the house dust mite allergen or poly(I:C). RNA expression profiles of cytokines were established using q-PCR and protein levels were determined in supernatants with ELISA. Results: The shRNA-mediated gene silencing reduced expression levels of EGR-1 by 92% (p <0.0001) and of DUSP-1 by 76% (p <0.0001). Both mutant cells lines showed an increased and prolonged response to the HDM allergen. The mRNA induction of IL-6 was 4.6 fold (p = 0.02) and 2.4 fold higher (p = 0.01) in the EGR-1 and DUSP-1 knock-down, respectively when compared to the induced levels in the control cell line. For IL-8, the induction levels were 4.6 fold (p = 0.01) and 13.0 (p = 0.001) fold higher. The outcome was largely similar, yet not identical at the secreted protein levels. Furthermore, steroids were able to suppress the poly(I:C) induced cytokine levels by 70-95%. Conclusions: Deregulation of EGR-1 and/or DUSP-1 in nasal epithelium could be responsible for the prolonged activated transcriptional state observed in vivo in allergic disease. This could have clinical consequences as cytokine levels after the steroid treatment in EGR-1 or DUSP-1 knock-down remained higher than in the control cell line. (C) 2015 Elsevier Ltd. All rights reserved
U2 - https://doi.org/10.1016/j.molimm.2014.12.011
DO - https://doi.org/10.1016/j.molimm.2014.12.011
M3 - Article
C2 - 25638726
SN - 0161-5890
VL - 65
SP - 43
EP - 50
JO - Molecular immunology
JF - Molecular immunology
IS - 1
ER -