Enhanced interleukin-1β production of PBMCs from patients with gout after stimulation with Toll-like receptor-2 ligands and urate crystals.docVIP

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Enhanced interleukin-1β production of PBMCs from patients with gout after stimulation with Toll-like receptor-2 ligands and urate crystals.doc

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Enhanced interleukin-1β production of PBMCs from patients with gout after stimulation with Toll-like receptor-2 ligands and urate crystals

Mylonaetal.ArthritisResearchTherapy2012,14:R158 /content/14/4/R158 RESEARCH ARTICLE OpenAccess Enhancedinterleukin-1bproductionofPBMCs frompatientswithgoutafterstimulationwith Toll-likereceptor-2ligandsanduratecrystals EleniEMylona1*,MariaMouktaroudi1,TaniaOCrisan2,3,StamatoulaMakri1,AikateriniPistiki1,MariannaGeorgitsi1, AthinaSavva1,MihaiGNetea2,3,JosWMvanderMeer2,3,EvangelosJGiamarellos-Bourboulis1,2,3 and LeoABJoosten2,3 Abstract Introduction:Monosodiumuratemonohydrate(MSU)crystalssynergizewithvarioustoll-likereceptor(TLR)ligands toinducecytokineproductionviaactivationoftheNOD-likereceptor(NLR)family,pyrindomain-containing 3(NLPR3)inflammasome.Thishasbeendemonstratedinvitrousinghumancelllinesormonocytesofhealthy volunteers.Inthepresentstudy,wehaveinvestigatedtheeffectofMSUcrystalsandoftheircombinationwithTLR ligandsinperipheralbloodmononuclearcells(PBMC)ofpatientswithgout. Methods:PBMCsfrom18patientswithprimarygoutand12healthydonorswereexposedtoMSUcrystalsinthe presenceorabsenceofsaturatedfattyacidC18:0(freefattyacid,TLR2ligand),palmitoyl-3-cystein(Pam3Cys,TLR1/2 ligand)andfibroblaststimulatingfactor-1(FSL-1,TLR2/6ligand).ProductionofIL-1b,IL-6,IL-8,IL-17andtumor necrosisfactoralpha(TNFa)wasdeterminedbyELISA.mRNAtranscriptsofIL-1bweremeasuredbyreal-timePCR. Results:MSUcrystalsalonefailedtoinduceIL-1b,IL-6orTNFainbothpatientsandcontrolgroups,butastronger synergybetweenMSU/PamCysandMSU/C18:0fortheinductionofIL-1bwasfoundinpatientswithgout 3 comparedtohealthycontrols.IL-6,butnotIL-8,followedthekineticsofIL-1b.Noproductionoftheneutrophil- recruitingIL-17wasdetectableafterstimulationofthepatients’PBMCswithMSUinboththepresenceorabsenceof TLRligands.NochangeofgenetranscriptsofIL-1bafterstimulationwithMSUandPam CysorwithMSUandC18:0 3 wasfound.ApositivecorrelationwasfoundbetweensynergyinIL-1bproductionfromPBMCsofpatientsbetween C18:0andMSUcrystals,aswellastheannualnumberofattacksofacutegoutyarthritis(rs:+0.649,P :0.022). Conclusions:ThesynergybetweenMSUcry