ORMDL3 regulates poly I:C induced inflammatory responses in airway epithelial cells

Abstract Background Oroscomucoid 3 (ORMDL3) has been linked to susceptibility of childhood asthma and respiratory viral infection. Polyinosinic-polycytidylic acid (poly I:C) is a synthetic analog of viral double-stranded RNA, a toll-like receptor 3 (TLR3) ligand and mimic of viral infection. Methods...

Full description

Bibliographic Details
Main Authors: Gemma Laura, Yi Liu, Kieran Fernandes, Saffron A. G. Willis-Owen, Kazuhiro Ito, William O. Cookson, Miriam F. Moffatt, Youming Zhang
Format: Article
Language:English
Published: BMC 2021-05-01
Series:BMC Pulmonary Medicine
Subjects:
Online Access:https://doi.org/10.1186/s12890-021-01496-5
id doaj-8a49f98f2be248f69b70ec796017d84a
record_format Article
spelling doaj-8a49f98f2be248f69b70ec796017d84a2021-05-23T11:27:46ZengBMCBMC Pulmonary Medicine1471-24662021-05-0121111310.1186/s12890-021-01496-5ORMDL3 regulates poly I:C induced inflammatory responses in airway epithelial cellsGemma Laura0Yi Liu1Kieran Fernandes2Saffron A. G. Willis-Owen3Kazuhiro Ito4William O. Cookson5Miriam F. Moffatt6Youming Zhang7National Heart and Lung Institute, Imperial College LondonNational Heart and Lung Institute, Imperial College LondonNational Heart and Lung Institute, Imperial College LondonNational Heart and Lung Institute, Imperial College LondonNational Heart and Lung Institute, Imperial College LondonNational Heart and Lung Institute, Imperial College LondonNational Heart and Lung Institute, Imperial College LondonNational Heart and Lung Institute, Imperial College LondonAbstract Background Oroscomucoid 3 (ORMDL3) has been linked to susceptibility of childhood asthma and respiratory viral infection. Polyinosinic-polycytidylic acid (poly I:C) is a synthetic analog of viral double-stranded RNA, a toll-like receptor 3 (TLR3) ligand and mimic of viral infection. Methods To investigate the functional role of ORMDL3 in the poly I:C-induced inflammatory response in airway epithelial cells, ORMDL3 knockdown and over-expression models were established in human A549 epithelial cells and primary normal human bronchial epithelial (NHBE) cells. The cells were stimulated with poly I:C or the Th17 cytokine IL-17A. IL-6 and IL-8 levels in supernatants,  mRNA levels of genes in the TLR3 pathway and inflammatory response from cell pellets were measured. ORMDL3 knockdown models in A549 and BEAS-2B epithelial cells were then infected with live human rhinovirus (HRV16) followed by IL-6 and IL-8 measurement. Results ORMDL3 knockdown and over-expression had little influence on the transcript levels of TLR3 in airway epithelial cells. Time course studies showed that ORMDL3-deficient A549 and NHBE cells had an attenuated IL-6 and IL-8 response to poly I:C stimulation. A549 and NHBE cells over-expressing ORMDL3 released relatively more IL-6 and IL-8 following poly I:C stimulation. IL-17A exhibited a similar inflammatory response in ORMDL3 knockdown and over-expressing cells, but co-stimulation of poly I:C and IL-17A did not significantly enhance the IL-6 and IL-8 response. Transcript abundance of IFNB following poly I:C stimulation was not significantly altered by ORMDL3 knockdown or over-expression. Dampening of the IL-6 response by ORMDL3 knockdown was confirmed in HRV16 infected BEAS-2B and A549 cells. Conclusions ORMDL3 regulates the viral inflammatory response in airway epithelial cells via mechanisms independent of the TLR3 pathway.https://doi.org/10.1186/s12890-021-01496-5ORMDL3Poly I:CTLR3Epithelial cellInflammatory response
collection DOAJ
language English
format Article
sources DOAJ
author Gemma Laura
Yi Liu
Kieran Fernandes
Saffron A. G. Willis-Owen
Kazuhiro Ito
William O. Cookson
Miriam F. Moffatt
Youming Zhang
spellingShingle Gemma Laura
Yi Liu
Kieran Fernandes
Saffron A. G. Willis-Owen
Kazuhiro Ito
William O. Cookson
Miriam F. Moffatt
Youming Zhang
ORMDL3 regulates poly I:C induced inflammatory responses in airway epithelial cells
BMC Pulmonary Medicine
ORMDL3
Poly I:C
TLR3
Epithelial cell
Inflammatory response
author_facet Gemma Laura
Yi Liu
Kieran Fernandes
Saffron A. G. Willis-Owen
Kazuhiro Ito
William O. Cookson
Miriam F. Moffatt
Youming Zhang
author_sort Gemma Laura
title ORMDL3 regulates poly I:C induced inflammatory responses in airway epithelial cells
title_short ORMDL3 regulates poly I:C induced inflammatory responses in airway epithelial cells
title_full ORMDL3 regulates poly I:C induced inflammatory responses in airway epithelial cells
title_fullStr ORMDL3 regulates poly I:C induced inflammatory responses in airway epithelial cells
title_full_unstemmed ORMDL3 regulates poly I:C induced inflammatory responses in airway epithelial cells
title_sort ormdl3 regulates poly i:c induced inflammatory responses in airway epithelial cells
publisher BMC
series BMC Pulmonary Medicine
issn 1471-2466
publishDate 2021-05-01
description Abstract Background Oroscomucoid 3 (ORMDL3) has been linked to susceptibility of childhood asthma and respiratory viral infection. Polyinosinic-polycytidylic acid (poly I:C) is a synthetic analog of viral double-stranded RNA, a toll-like receptor 3 (TLR3) ligand and mimic of viral infection. Methods To investigate the functional role of ORMDL3 in the poly I:C-induced inflammatory response in airway epithelial cells, ORMDL3 knockdown and over-expression models were established in human A549 epithelial cells and primary normal human bronchial epithelial (NHBE) cells. The cells were stimulated with poly I:C or the Th17 cytokine IL-17A. IL-6 and IL-8 levels in supernatants,  mRNA levels of genes in the TLR3 pathway and inflammatory response from cell pellets were measured. ORMDL3 knockdown models in A549 and BEAS-2B epithelial cells were then infected with live human rhinovirus (HRV16) followed by IL-6 and IL-8 measurement. Results ORMDL3 knockdown and over-expression had little influence on the transcript levels of TLR3 in airway epithelial cells. Time course studies showed that ORMDL3-deficient A549 and NHBE cells had an attenuated IL-6 and IL-8 response to poly I:C stimulation. A549 and NHBE cells over-expressing ORMDL3 released relatively more IL-6 and IL-8 following poly I:C stimulation. IL-17A exhibited a similar inflammatory response in ORMDL3 knockdown and over-expressing cells, but co-stimulation of poly I:C and IL-17A did not significantly enhance the IL-6 and IL-8 response. Transcript abundance of IFNB following poly I:C stimulation was not significantly altered by ORMDL3 knockdown or over-expression. Dampening of the IL-6 response by ORMDL3 knockdown was confirmed in HRV16 infected BEAS-2B and A549 cells. Conclusions ORMDL3 regulates the viral inflammatory response in airway epithelial cells via mechanisms independent of the TLR3 pathway.
topic ORMDL3
Poly I:C
TLR3
Epithelial cell
Inflammatory response
url https://doi.org/10.1186/s12890-021-01496-5
work_keys_str_mv AT gemmalaura ormdl3regulatespolyicinducedinflammatoryresponsesinairwayepithelialcells
AT yiliu ormdl3regulatespolyicinducedinflammatoryresponsesinairwayepithelialcells
AT kieranfernandes ormdl3regulatespolyicinducedinflammatoryresponsesinairwayepithelialcells
AT saffronagwillisowen ormdl3regulatespolyicinducedinflammatoryresponsesinairwayepithelialcells
AT kazuhiroito ormdl3regulatespolyicinducedinflammatoryresponsesinairwayepithelialcells
AT williamocookson ormdl3regulatespolyicinducedinflammatoryresponsesinairwayepithelialcells
AT miriamfmoffatt ormdl3regulatespolyicinducedinflammatoryresponsesinairwayepithelialcells
AT youmingzhang ormdl3regulatespolyicinducedinflammatoryresponsesinairwayepithelialcells
_version_ 1721429705198927872