IL‐36γ is secreted in microparticles and exosomes by lung macrophages in response to bacteria and bacterial components

Journal of Leukocyte Biology: An Official Publication of the Reticuloendothelial Society

关键词: innate immune response;    pneumonia;    nonclassic secretion;    cytokines;   

DOI  :  10.1189/jlb.4A0315-087R

学科分类：生理学

来源: Federation of American Societies for Experimental Biology

【 摘 要 】

Interleukin‐36isafamilyofnovelinterleukin‐1‐likeproinflammatorycytokinesthatarehighlyexpressedinepithelialtissuesandseveralmyeloid‐derivedcelltypes.Likethoseofclassicinterleukin‐1cytokines,thesecretionmechanismsofinterleukin‐36arenotwellunderstood.Interleukin‐36γsecretionindermalepithelialcellsrequiresadenosine5'‐triphosphate,whichsuggestsanonclassicalmechanismofsecretion.Inthisstudy,murinepulmonarymacrophagesandhumanalveolarmacrophagesweretreatedwithrecombinantpathogen‐associatedmolecularpatterns(intactbacteria:KlebsiellapneumoniaeorStreptococcuspneumoniae).Celllysateswereanalyzedformessengerribonucleicacidbyquantitativereal‐timepolymerasechainreaction,andconditionedmediumwasanalyzedforinterleukin‐36γbyenzyme‐linkedimmunosorbentassay,withorwithoutsonication.Inaddition,conditionedmediumwasultracentrifugedat25,000gand100,000g,toisolatemicroparticlesandexosomes,respectively,andinterleukin‐36γproteinwasassessedineachfractionbyWesternblotanalysis.Interleukin‐36γmRNAwasinducedinbothmurineandhumanlungmacrophagesbyavarietyofpathogen‐associatedmolecularpatterns,aswellasheat‐killedandliveKlebsiellapneumoniaeandStreptococcuspneumoniae,andinductionoccurredinamyeloiddifferentiationresponsegene88–dependentmanner.Secretionofinterleukin‐36γproteinwasenhancedbyadenosine5'‐triphosphate.Furthermore,extracellularinterleukin‐36γproteindetectionwasmarkedlyenhancedbysonicationtodisruptmembrane‐boundstructures.Interleukin‐36γproteinwasdetectedbyWesternblotinmicroparticlesandexosomefractionsisolatedbyultracentrifugation.Interleukin‐36γwasinducedandsecretedfromlungmacrophagesinresponsetoGram‐negativeand‐positivebacterialstimulation.Theresultssuggestthatinterleukin‐36γissecretedinanon‐Golgi–dependentmannerbylungmacrophagesinresponsetoGram‐positiveand‐negativebacterialchallenge...

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