Chronic IL9 and IL-13 Exposure Leads to an Altered Dferentiation of Ciliated Cells in a Well-Dferentiated Paediatric Bronchial Epithelial Cell Model.docVIP

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Chronic IL9 and IL-13 Exposure Leads to an Altered Dferentiation of Ciliated Cells in a Well-Dferentiated Paediatric Bronchial Epithelial Cell Model.doc

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Chronic IL9 and IL-13 Exposure Leads to an Altered Dferentiation of Ciliated Cells in a Well-Dferentiated Paediatric Bronchial Epithelial Cell Model

ChronicIL9andIL-13ExposureLeadstoanAltered DifferentiationofCiliatedCellsinaWell-Differentiated PaediatricBronchialEpithelialCellModel JeremyC.Parker1.,SurendranThavagnanam4.,GrzegorzSkibinski1,JeremyLyons3,JenniferBell1, LiamG.Heaney1*,MichaelD.Shields1,2 1CentreforInfectionandImmunity,Queen’sUniversityBelfast,Belfast,NorthernIreland,2RoyalBelfastHospitalforSickChildren,BelfastHealthandSocialCareTrust, Belfast,NorthernIreland,3TheRoyalHospitals,BelfastHealthandSocialCareTrust,Belfast,NorthernIreland,4PaediatricDepartment,UniversityMalayaMedicalCentre, KualaLumpar,Malaysia Abstract Asthmaisachronicinflammatorydiseasecharacterisedbyairwaysremodelling.InmousemodelsIL-9andIL-13havebeen implicatedinairwaysremodellingincludingmucushypersecretionandgobletcellhyperplasia.Theirrole,especiallythatof IL-9,hasbeenmuchlessstudiedinauthentichumanexvivomodelsofthebronchialepitheliumfromnormalandasthmatic children.WeassessedtheeffectsofIL-9,IL-13andanIL-9/IL-13combination,duringdifferentiationofbronchialepithelial cells from normal (n=6) and asthmatic (n=8) children. Cultures were analysed for morphological markers and factors associated with altered differentiation (MUC5AC, SPDEF and MMP-7). IL-9, IL-9/IL-13 combination and IL-13 stimulated bronchialepithelialcellsfromnormalchildrenhadfewerciliatedcells[14.8%(SD8.9),p=0.048,12.4(SD6.1),p=0.016and 7.3%(SD6.6),p=0.031]respectivelycomparedwithunstimulated[(21.4%(SD9.6)].IL-9stimulationhadnoeffectongoblet cellnumberineithergroupwhereasIL-9/IL-13combinationandIL-13significantlyincreasedgobletcellnumber[24.8%(SD 8.8), p=0.02), 32.9% (SD 8.6), p=0.007] compared with unstimulated normal bronchial cells [(18.6% (SD 6.2)]. All stimulations increased MUC5AC mRNA in bronchial epithelial cells from normal children and increased MUC5AC mucin secretion. MMP-7 localisation was dysregulated in normal bronchial epithelium stimulated with Th2 cytokines which resembledtheunstimulatedbronchialepitheliumofasthmaticchildren.Allstimulationsresultedin