Deciphering Innate Immunity And Tolerance From Single Molecules To Systemic Levels – NKTOLERANCE

Selfversusnon-selfdiscrimination isacentralthemein biology,extending fromplantsto vertebrates.Naturalkiller(NK)cellsin themammalian immunesystemconstitutean experimentalmodelsystemforinvestigating themolecularand cellularmechanismsinvolvedin thesecomplex biologicalprocesses.Indeed,NKcellsaretolerantto normalselftissues,butkilland producecytokinesin responseto cellsin distress(tumors,infected cells...).Studiesinhumansand micehaveshown thatself-recognition isrequired forNKcellsto acquirefunctionalcompetence.However,littleisknownaboutthisdevelopmentalprocess.Wethereforeproposeto elucidatethemechanismsofNKcell"education"to selfversusnon-selfdiscrimination.Understanding such complex system,fromthemolecularscaleto higherleveltissuearchitectureand beyond,to thephysiology ofwholeorganisms,requiresmultipleapproachesatvariouslevels.Wethusproposeto work upfromthewholeorganismto themolecularscale.In particular,newmousemutantswillbegenerated by randomgermlinemutagenesisusing ENU(N-ethyl-N-nitrosourea).Wewillalsotry to understand thesystemasawhole,usinggenome-widetranscriptomeanalysisand NKcellimaging in tissues.Finally,thereal-timeobservation ofprotein dynamicsin living cellsusing aFCS(FluorescenceCorrelation Spectroscopy)method willallowto measureparameters,suchasthenumberofmolecules,diffusion coefficients,spatialdistributionsand temporalfluctuations,essentialforthegeneration ofquantitativemodels.By determining themechanismsallowing bothNKcellself-toleranceand theirability torespond todiseased cells,weshould improveourunderstanding ofbiologicalprocessesthatmay operatebroadly in thevertebrateinnateimmunesystem.Thisinter-disciplinary projectshould also contributetodefinetherationalebasisofmanipulating NKcellsin ongoing and forthcoming innovativeanti-cancerclinicaltrials