IMMUNImmunology Meeting - Strasbourg
2016
Organized by: MUELLER GC (CNRS- IBMC) – CHAN S (CNRS-INSERM) – PhD students : EL MDAWAR M – EL HACHEM C – ALLOUSH F
Programmedoctoral internationalInternationaldoctoralprogramme
The First Immunology Meeting of Strasbourg is organized by researchers and PhD candidates of the university of Strasbourg. It aims to present scientific activity and to foster exchange of ideas and collaborations. Indeed, this meeting focuses on basic research that may alter our understanding of existing concepts in immunology.
Different research institutes and structures of the university on Strasbourg will present their research activities organized into 4 themes:
1. Adaptive and innate immunity 2. Hematopoiesis and development 3. Inflammation regulatory mechanisms 4. Therapy
Participants include post-doc researchers and PhD candidates from the following organizations:
− Institut de Biologie Moléculaire et Cellulaire "IBMC" − Institut de Génétique et de Biologie Moléculaire et cellualire
"IGBMC" − Etablissement Français de Sang "EFS" − Institute of Immunology of Strasbourg - Faculty of Medecine of
Strasbourg − Faculty of Pharmacy of Strasbourg
The organizing Committee:
− Christopher Mueller (DR, CNRS UPR 3572, IBMC) − Susan Chan (DR, IGBMC, Illkirch) − Carole El Hachem (PhD Student, IGBMC, Illkirch) − Marie-Belle El Mdawar (PhD Student, UMR 949, EFS-ALCA) − Farouk Allouch (PhD Student, CNRS UPR 3572, IBMC)
1
PROGRAM
8:30 – 9:00 Breakfast
9:00 – 9:15 Welcome Speech
9:15 – 10:45 Adaptive & Innate Immunity
Chairs: Susan Chan & Christopher Mueller Alexandre Mariotte (2nd year PhD student), supervisor: Philippe Georgel DICER and inflammation Isaque João da Silva de Faria (2nd year PhD Student), supervisors: Jean-Luc Imler and Joao T. Marques (UFMG Belo-Horizonte) DNA virus sensing through viral dsRNA recognition by Drosophila innate immune system
Delphine Bouis (2nd year PhD student), supervisor: Pauline Soulas-Sprauel Role of STING gain of function in immunity
Ramzi Nehmar (4th year PhD student), supervisor: Philippe Georgel Role of plasmacytoid dendritic cells in mouse models of experimental arthritis Marie-Belle El Mdawar (3rd year PhD student), supervisor: Henri de la Salle Cells implicated in the development of Transfusion-Related Acute Lung Injury Carole El Hachem (4th year PhD student), supervisor: Mei Li Basophils in allergic skin inflammation
10:45 – 11:15 Coffee Break – offered by StrasAIR association
11:15 – 12:30 Development and hematopoiesis talks Chair: Marie-Belle El Mdawar
Alicia Aguilar (4th year PhD student), supervisor: Catherine Léon Mimicking the bone marrow environment in 3D culture, a key to better understand megakaryopoiesis
Camille Jost (2nd year PhD student), supervisors: François Lanza et Nathalie Brouard Role of the cellular microenvironment in megakaryocyte differentiation Quentin Muller (3rd year PhD student), supervisor: Vincent Flacher Reconstruction of human immunocompetent and innervated human skin Romain Veber (2nd year PhD student), supervisor: Hélène Dumortier Lymphoid neogenesis in kidneys during lupus: fundamental mechanisms and therapeutic tracks
2
Gaëtan Maurer (3rd year PhD student), supervisors: Philippe Kastne et Céline Charvet Role of Ikaros in Th17 cell polarization
12:30 – 14:00 Buffet LUNCH (offered by Medalis LabEx)
14:00 – 14:45 Inflammation I Chair: Carole El Hachem
Blandine Maitre (University of Toronto/UMR_S949 et EFS-ALCA) Role of Vitamin A and LTβR in the homeostasis of splenic dendritic cells Camille Balbinot (4th year PhD student), supervisor: Isabelle Duluc Non-cell-autonomous tumor suppressor activity of the Cdx2 homeobox gene in the gut Matthieu Sawaf (3rd year PhD student), supervisor: Fanny Monneaux Role of BTLA in human systemic lupus erythematosus
14:45 – 15:15 Coffee & Tea Break
15:15 – 16:00 Inflammation II Chair: Carole El Hachem
Mélanie Chypre (3rd year PhD student), supervisor: Christopher Mueller Regulation of lymph node macrophage differentiation by RANKL
Virginia Delgado (Post-doc), supervisor: Pauline SOULAS-SPRAUEL Searching partners of Trib1 in B cells
Ruicheng Wei (Post-doc), supervisor: Mei Li Study of type 2 immunity in atopic dermatitis
16:00 – 16:15 Coffee & Tea Break
16:15 – 17:15 Therapy Chair: Farouk Alloush
Fengjuan Wang (Post-doc), supervisor: Sylviane Muller P140 peptide, a heat shock protein inhibitor, targets lysosomes for lupus therapy
Baihui Li (3rd year PhD student), supervisor: Sylviane Muller P140 as a modulator of autophagy in Sjögren's syndrome
François Daubeuf (Post-doc), supervisor: Nelly Frossard Mechanisms of action of an CXCL12 inhibitor in the anti-inflammation of asthma
Celia Jacoberger-Foissac (3rd year PhD student), supervisors: Béatrice Heurtault, Sylvie Fournel Conception and evaluation of liposomes-based therapeutic vaccines against cancer in a murine model
17:15 – 17:20 Concluding Remarks
DICERandinflammation
A.Mariotte1|,A.DeCauwer1,R.Veber2,G.Alsaleh1,R.Nehmar1,H.Dumortier2,S.Muller2andP.Georgerl1*.
1INSERMU1109,StrasbourgUniversity;2IBMC,StrasbourgUniversity
Abstract
DICER,anRNaseIIIfamilyenzyme,isimplicatedinthebiogenesisofmicroRNAs,agroupofbonafideRNA-silencing
components demonstrated as almost linked to every biological function. Although microRNAs are crucial in
inflammatory contexts, DICER also seems to participate in innate immune modulations through its role in the
turnoverofpotentiallyimmune-activatorsequencessuchasAluRNAs.Inaddition,recentworkshavealsosuggested
thatsuchregulationsofAluRNAbreakdowncouldbeinvolvedinthepathogenesisofsystemiclupuserythematosus
(SLE),anautoimmunedisease.OuraimistostudytheroleofDICERinthissettingbyinducingimiquimod-induced
SLE-likediseasetoDICER-deficient(Dicerd/d)or-sufficientmice.Wethenmonitoredrenaldysfunctionbyquantifying
proteinuriawithurinarystripsandarticular/dermatologicsymptomswereevaluatedbyvisualscoring.Wenoticed
thatDicerd/d
micetendtodevelopamoreseverelupus-likediseasethanthewild-typecounterpart(increasedrenal
dysfunction,articularsymptomsandtypicalcutaneousrashes).Thosefindingscouldbeofparticularinterestfora
betterunderstandingofSLEphysiopathology.
|Speaker
*Correspondence:[email protected]
2
DNAvirussensingthroughviraldsRNArecognition
byDrosophilainnateimmunesystem
IsaqueJoãodaSilvadeFaria1,2|,EricRobertoGuimarãesRochaAguiar1,JoãoTrindadeMarques1,2andJean-LucImler2*
1DepartmentofBiochemistryandImmunology,UniversidadeFederaldeMinasGerais,Brazil;2CNRS-UPR9022,
InstitutdeBiologieMoléculaireetCellulaire,UniversityofStrasbourg,France
Abstract
Virus-derivedsmallinterferingRNA(vsiRNA)mediatingviralcontrolisessentialforantiviraldefenseininvertebrates.
InDrosophilamodel,vsiRNAsaregeneratedthroughDicer-2processingofviraldouble-stranded(ds)RNAandloaded
intoArgonaute2forviraltranscriptsilencing.InfectionbyInvertebrateiridescentvirus6(IIV6),alargeDNAvirus,
triggerstheproductionofvsiRNAsinDrosophilacellsandadultflies.TheanalysisofsmallRNA-seqdatafromIIV6-
infectedcells showsvsiRNAsarederived fromgenomichotspotswithhighvsiRNAdensity.Within these regions,
vsiRNAswerealignedbothingenicandintergenicregions.UsingfeaturesofDcr-2processingandvsiRNAcoverage,
wesuggest thatvsiRNAsderived from IIV6genomeareproducedas longdsRNAprecursors. Inaddition,wealso
detected sense and antisense transcripts from these dsRNAprecursors. Interestingly, inhibition of the viral RNA
polymerase does not impair production of IIV6-derived siRNAs. Altogether, our results suggest that a host RNA
polymerasetranscribesviralRNAsuchthatdsRNAisproduced,thusactivatingDicer-2andantiviralinnateimmunity.
|Speaker
*Correspondence:[email protected]
3
RoleofSTINGgainoffunctioninimmunity
DelphineBouis1|,AnneSoley1,2,ThierryMartin1,2,Anne-SophieKorganow1,2,PaulineSoulas-Sprauel1,2,in
collaborationwithFredericRieux-Laucat3(Imagine,Paris)andANRLumugenegroup3
1ICT,UPR3572,IBMC,Strasbourg;2HôpitauxUniversitairedeStrasbourg,NouvelHôpitalCivil,Strasbourg3ImagineInstitute,HôpitalNecker,Paris
Abstract
Ourcollaborator,DrFrédéricRieux-Laucat(Imagineinstitute,Paris;ANRLumugene),hasstudiedfamilialcasesof
systemiclupuserythematosus(SLE).Inoneofthesemultiplexfamilies,aheterozygousmutation(V155M)was
identifiedinTMEM173/STINGgene,encodingSting,andleadingtothedevelopmentofanautoinflammatory
diseasewithvasculopathyandpulmonaryfibrosis,describedasaninterferonopathy,andassociatedtoSLE-like
symptoms.InordertobetterunderstandtheconsequencesofSTINGgainoffunctioninthedevelopmentofthis
complexinflammatoryphenotype,wehavedevelopedanewmousemodelconsistinginaknock-in(KI)modelfor
thecorrespondingpointmutation(V154M)inmurineStinggene,byCrisprCas9technology.Thesemicestrikingly
developastrongphenotypewithlowSCIDimmunodeficiency(affectingT,BandNKcells)inaspecificpathogen
free(SPF)animalfacility.Analysesareinprogresstodissectthemechanismsofthisimmunodeficiency.Inaddition,
inordertounderstandtheroleofenvironmentintheobservedphenotype,wegeneratedourmousemodelin
specificandopportunisticpathogenfree(SOPF)conditions.WealsostartedtointercrossourmicewithIFNARKO
mice(deficientfortheIFN-Ireceptor)inordertochecktheinvolvementoftypeIIFNs.
References:
Jeremiahetal.,InheritedSTING-activatingmutationunderliesafamilialinflammatorysyndromewithlupus-likemanifestations.J.Clin.Invest,2014;1-5.
Liuetal.,ActivatedSTINGinaVascularandPulmonarySyndrome.N.Engl.J.Med.,2014;371:507-518.
|Speaker
4
Roleofplasmacytoiddendriticcellsinmousemodelsofexperimentalarthritis
R.Nehmar1,G.Alsaleh1,B.Voisin2,V.Flacher2,P.Kastner3,S.Bluml4,C.Muller2,S.Bahram1andP.Georgel1
1INSERMU1109,Strasbourg;2IBMC,Strasbourg;3IGBMC,Illkirch;4MedicalUniversity,Vienna,Austria
Abstract
Plasmacytoiddendriticcells(pDCs)aremajortype-IIFNproducersfollowingactivation.Theyplayanimportantrole
intheinitiationoftheinflammatoryresponseandparticipatetotheetiologyofseveralchronicdiseases.However,
anti-inflammatory actions of type-I IFNwere also considered, especially IFN-β. The aimof ourwork is to better
characterizetheroleofpDCsinRAusingmousemodelstoclarifythesecontradictoryobservations.Arthritisinmice
was inducedbyarthritogenic(K/BxN)serumtransferorupon injectionofheterologouscollagen(CIA).Symptoms
were evaluated by visual scoring andmeasurements of the thickness of the joints. Cellular infiltrates and pDCs
depletionwereanalyzedbyFACS,cytokinesexpressionwithELISAandRTqPCRandboneerosionwasevaluatedby
histological staining (TRAP). A mouse genetic model (IkarosL/L
) of pDC deficiency showed exacerbation of
inflammatoryandarthriticsymptomsafterarthritogenicserumtransfer;thiswasalsoobservedintwoothersmouse
modelsofpDCsdepletion.Next,weusedtopicalapplicationofaTLR7agonistwhichinducespDCsrecruitmentatthe
inflammatoryarthriticjoints.ThistreatmentreducesarticularinflammationinK/BxNandCIAarthritismodels.Our
resultssuggestthatpharmacologicaltargetingofpDCscouldhaveabeneficialeffectinarthritis.
|Speaker
*Correspondence:[email protected]
5
CellsimplicatedinthedevelopmentofTransfusion-RelatedAcuteLungInjury
Marie-BelleEl-Mdawar1,2,3,4|,StéphanieMagnenat1,2,3,4,FlorynaLefebvre1,2,3,4,ChristianGachet1,2,3,4,BéatriceHechler1,2,3,4,HenriDeLaSalle1,2,3,4*
1UMR_S949,INSERM,Strasbourg;2EtablissementFrançaisduSang-Alsace(EFS-Alsace),Strasbourg;3UniversitédeStrasbourg;4FédérationdeMédecineTranslationnelledeStrasbourg(FMTS)
Abstract
Antibody-mediated TRALI (Transfusion-Related Acute Lung Injury) is considered a rare remaining cause of
transfusion-relatedmortality,underscoringtheneedtounderstandthefactorsimplicatedinthedisease.TRALIcan
beprovokedbythepresenceofallogeneicantibodies–e.g.anti-HLAclassI−inthetransfusedbloodproducts.A
mousemodelofantibody-mediatedTRALIhasbeendevelopedintheH-2dbackground.Inthisexperimentalmodel,
platelets and neutrophils have been reported to be implicated in the development of TRALI. However these
conclusionsarenotsupportedbyresultsfromourlaboratoryandothers.Rather,workshavesuggestedakeyroleof
monocytes/macrophages and the production of reactive oxygen species (ROS).Which monocytes/macrophages
populationsandwhichROS-producingcellsareinvolvedneedtobeclarified.Wedepletedspecificmonocytesand
macrophagesusingdifferentmethods.ExperimentsindicatethatTRALIdependsonthepresenceofmacrophages.
Experimentsareongoingtodeterminewhichmacrophagessub-populationsareinvolved.Inaddition,weassessed
the production of ROS in pulmonary cells. We found that all lung cells produce ROS during TRALI, whereas
monocytes/macrophagesdepletion inhibitsthisproductionbyallcell types.Theseresultssuggestawiderroleof
cellsandROSinTRALI.
|Speaker
*Correspondence:[email protected]
6
Basophilsinallergicskininflammation
CaroleELHachem1|andMeiLi1*
1DepartmentofFunctionalGenomicandCancer,IGBMC,1rueLaurentFries,67404Illkirch-Graffenstaden,France
Abstract
Basophils areone typeof circulatinggranulocytes thataccount for1%ofblood leucocytes. They representone
characteristiccellularcomponentinallergicskininflammation,buttheircontributiontoimmuneresponsesremain
largelyundefined,mainlyduetothelackingofreagents/toolsfortrackingandfunctionallystudyingbasophilsinvivo.
Recently, antibodies specifically detecting basophils, and reagents/tools for depleting basophils have been
developed.Iwilldiscusshowwecombinethesereagents/toolswithmousemodelsandcellularandmolecularbiology
approaches,toinvestigatebasophilrecruitmentandactivation,aswellastheircrosstalkandinterplaywithother
immunecells,includingTcells,eosinophilsandneutrophilsinallergicskininflammation.
|Speaker
*Correspondence:[email protected]
7
Mimickingthebonemarrowenvironmentin3Dculture,akeytobetter
understandmegakaryopoiesis
AliciaAguilar1,2,3,4•,FabienPertuy1,2,3,4,AnitaEckly1,2,3,4,CatherineStrassel1,2,3,4,DominiqueCollin5,Christian
Gachet1,2,3,4,FrançoisLanza1,2,3,4,CatherineLeon1,2,3,4*1UMR_S949,INSERM,Strasbourg;2EtablissementFrançaisduSang-Alsace(EFS-Alsace),Strasbourg;3UniversitédeStrasbourg;4FédérationdeMédecineTranslationnelledeStrasbourg(FMTS);5InstitutCharlesSadron,UPR22,
Strasbourg.
Abstract
Plateletsarevitalbloodelementsastheystopbleedingandpreventhemorrhages.Plateletsarisefrom
giantbonemarrowcellscalledmegakaryocytes(MK),whichderivethemselvesfromthedifferentiationof
hematopoietic stem cell via the megakaryopoiesis process. The ultimate stage is the extension of
proplatelets through the sinusoid vessels to release platelets. The bone marrow environment is a
meshworkofextracellularmatrixproteinssurroundinghematopoieticcellsandthesoftesttissueofthe
body(~300Pa)[1].Itisnowrecognizethatcells“feel”stiffnessandtopographyoftheirlocalenvironment
andadapttheirmorphologyanddifferentiationprogrammbyaprocesscalledmechanotransduction[2].
To study the impact of stiffness and dimensionality onmegakaryopoiesis,we seed progenitor cells in
methylcellulose hydrogelmimicking the bonemarrow stiffness.We show that 3D culture contributes,
through the actomyosin and MKL1 mechanotransduction pathways, to a more favorable in vitro
environmentforMKdifferentiationwhichultimatelytranslatesintoincreasedproplateletproduction[3].
References:
[1]Choi,J.S.&Harley,B.aC.Thecombinedinfluenceofsubstrateelasticityandliganddensityontheviabilityandbiophysicalpropertiesofhematopoieticstemandprogenitorcells.Biomaterials33,4460–8(2012).[2]Engler,A.J.,Sen,S.,Sweeney,H.L.&Discher,D.E.Matrixelasticitydirectsstemcelllineagespecification.Cell126,677–89(2006).[3]Aguilar,A.etal.Importanceofenvironmentalstiffnessformegakaryocytedifferentiationandproplateletformation.Bloodblood–2016–02–699959(2016).doi:10.1182/blood-2016-02-699959
•Speaker
*Correspondence:[email protected]
8
Roleofthecellularmicroenvironmentinmegakaryocytedifferentiation
CamilleJost1,2,3,4●,NathalieBrouard1,2,3,4,CatherineStrassel1,2,3,4,ChristianGachet1,2,3,4andFrançoisLanza1,2,3,41UMR_S949,INSERM,Strasbourg;2EtablissementFrançaisduSang-Alsace(EFS-Alsace),Strasbourg;3Universitéde
Strasbourg;4FédérationdeMédecineTranslationnelledeStrasbourg(FMTS).
Abstract
Platelets are anucleate elements of the bloodwhich stop bleeding. They are produced in the bonemarrow by
megakaryocytes(MK)thatderivefromhematopoieticstemcells(HSC)duringaprocesscalledmegakaryopoiesis.A
cellularmicroenvironment composed of stromal cells interactswith hematopoietic progenitors to regulate their
proliferation and differentiation. During embryogenesis, the fetal liver (FL) is the site ofmajor proliferation and
expansionofhematopoieticcellsincludingMK.Weisolatedmultiplestromalcellularcomponentsfrommousefetal
liverandevaluatedtheircapacitytosupport/regulatemegakaryopoiesis.Hereinweco-culturedhumanHSCisolated
fromperipheralbloodwithphenotypicallydistinctmurineFLstromalcellsinalowcytokinemediumtoevaluatetheir
respective role inmegakaryopoiesis.We found that a stromal cell population, identified as CD45-TER119-CD31-
CD51+VCAM-1+PDGFRa-, supports the production of a large number of committed cells andMK fromHSC. The
mechanismsinvolvedinthisparticularinteractionremaintobeidentifiedwiththeperspectivetoimproveplatelet
productioninvitro.
●Speaker
9
Constructionofatridimensionalimmunocompetent,innervatedand
vascularizedhumanskinmodel
QuentinMuller1|,Marie-JoséeBeaudet2,EvelyneSchaeffer1,ChristopherMueller1,FrançoisBerthod2#
andVincentFlacher1*#
1Team«Immune-microenvironmentinteractionsinhealthanddisease»,CNRS-UPR3572/LabExMedalis«ImmunopathologieetChimieThérapeutique»,InstitutdeBiologieMoléculaireetCellulaire,France;2Laboratoired’OrganogénèseEXpérimentaledel’UniversitéLaval,CHUde
Québec-UniversitéLaval,Canada;#Equalcontribution.
Abstract
Immunereactionsintheskinareinitiatedbythecutaneousdendriticcells(DCs).Thepotentialsensitizingeffectofa
compoundcanbepredictedinvitrousinghumanbloodmonocytesdifferentiatedintoDCs(Mono-DCs)ormonocytic
celllines.However,thesesimplisticmodelsremaininaccuratebecausetheactivationofcutaneousDCsbysensitizers
maybetriggeredormodulatedbymicroenvironmentalinteractionswithmultipletypesofnon-immunecells[1].Our
goalistodevelopanimmunocompetenttissue-engineeredskin(TES)thatwillcombineDCswithallstructuraland
functionalelementoftheskin,i.e.anepidermalbarrierlaiduponadermiscontainingapseudo-vascularizationand
nociceptiveneurons[2].Collagen-chitosanlatticeswerefirstseededwithfibroblastsandendothelialcells,thenwith
precursorsofnervefibersderivedfromeitherhumaninducedpluripotentstemcellsormurineembryonicdorsal
rootganglia.Finally,weintroducedkeratinocytesandMono-DCs.Weobservedthatinsitudifferentiatedneurons
growaxonstowardstheepidermis.Moreover,Mono-DCssettledasexpectedbeneaththeepidermisandremained
sessileforseveralweeks.Inthenearfuture,weplantoevaluateinourmodelthesensitivityofDCs,innervationand
microvasculaturetochemicalcompoundsandotherdangersignals.
References:
[1]Riol-BlancoL,Ordovas-MontanesJ,PerroM,NavalE,ThiriotA,AlvarezD,etal.Nociceptivesensoryneuronsdriveinterleukin-23-mediatedpsoriasiformskininflammation.Nature.2014;510(7503):157-61.[2]CadauS,Leoty-OkombiS,PainS,BechetoilleN,Andre-FreiV,BerthodF.Invitroglycationofanendothelializedandinnervatedtissue-engineeredskintoscreenanti-AGEmolecules.Biomaterials.2015;51:216-25.
|Speaker
*Correspondence:[email protected]
10
Lymphoidneogenesisinkidneysduringlupus:fundamentalmechanismsand
therapeutictracks
RomainVeber1|,CaroleLeCoz1,SophiaLecomte1,FannyMonneaux1,KristinA.FentonandHélèneDumortier1*
1CNRS,UPR3572ImmunopathologieetChimieThérapeutique,InstitutdeBiologieMoléculaireetCellulaire,67084Strasbourg,France;2RNAandMolecularPathologyResearchGroup,InstituteofMedicalBiology,FacultyofHealthSciences,UniversityofTromsø,9037Tromsø,Norway
Abstract
TertiaryLymphoidOrgans(TLOs)arestructuressimilartolymphnodes,whichdevelopduringchronicinflammation.
They can be observed in pathological situations such as autoimmune diseases and participate to local harmful
immuneresponses.Ourlaboratoryworksonlupus,achronicandsystemicautoimmunediseasethatleadstomultiple
organ failures amongwhich kidney failure.We have demonstrated that TLOs are present in the kidneys of the
spontaneous NZB/W lupus mouse model. In this context, the goals of my current work are i) to elucidate the
mechanisms leadingtoTLOneogenesis,and2)todeveloptherapeuticstrategiesbasedonthetargetingofthese
mechanisms.First,smallleukocyteinfiltratesarevisualizedveryearlyinthekidneysofyoungNZB/Wmiceandinthe
absence of glomerular deposits and of detectable autoantibodies in the serum, suggesting that immune cell
infiltrationoccursbeforetheautoantibody-inducedkidneyinflammation.Moreover,amongthefirstcellsentering
kidneys,wefoundamajorityofactivated/memoryTcellsthatmaybetheinitiatorsofTLOformation.Altogether,
ourdatasuggestthatTLOneogenesistakesplaceataveryearlystageofdiseasedevelopmentandthatblockingT
cellinfiltrationcouldimpairTLOgenerationandpreventkidneydysfunction.
|Speaker
*Correspondence:[email protected]
11
InvestigatingtheroleofIkarosinTH17cellsGaëtanMaurer,1|CélineCharvet1*andPhilippeKastner1*.
1DepartmentofFunctionalGenomicandCancer,IGBMC,1rueLaurentFries,67404Illkirch-Graffenstaden,France
Abstract
CD4Thelper17(TH17)lymphocytesarecriticalplayersinthedefenseagainstextracellularbacteriaor
fungibuttheirderegulationcanleadtoautoimmuneinflammatorydiseasessuchasmultiplesclerosisor
rheumatoidarthritis.Understandingthemolecularmechanismsregulatingtheswitchfromnon-
pathogenictopathogenicTH17is,therefore,ofmajorimportance.Ikarosisazinc-fingertranscription
factor,havingacrucialfunctioninBandTcellmalignanciesanddifferentiationprocesses.Theobjective
ofourprojectistoexploretheroleofIkarosinTH17differentiation.Wewillpresentourresultsobtained
byglobalexperimentalapproaches:(i)transcriptomeand(ii)ChIP-seqanalysisonTH17cells.Thesedata
suggestthatIkaroscanimpactasetofgenesdefiningthepathogenicTH17signaturenotbydirectly
bindingontheirregulatorysequences,butratherviaanindirectmechanism.Together,theseglobal
approacheswillhelpusgainnewinsightsontheroleofIkarosinthegeneration,stabilityand
pathogenicityofTH17lymphocytes
|Speaker
*Correspondence:[email protected]@igbmc.fr
12
RoleofvitaminAandLTβRinthehomeostasisofsplenicdendriticcells
BlandineMaître1,AlbertNguyen1,TianSun1,MarieIrondelle2,JeanSalamero2andJenniferL.Gommerman1*
1DepartmentofImmunology,UniversityofToronto,Canada;2InstitutCurie,UMR144,Paris,France
Abstract
An efficient response to blood-borne antigens requires the appropriate homeostasis and positioning of splenic
dendriticcells(DCs).SplenicCD11b+CD8-ESAM+DCsarelocalizedwithinthemarginalzonebridgingchannels(MZ
BC),aregionwheretheTcellzoneconnectstotheredpulpandwhereDCscantakeupblood-borneantigens.Among
thefactorsinvolvedinDChomeostasis,weexamineheretherespectivecontributionofretinoicacid(RA),avitamin
Aderivative,andDC-intrinsicLymphotoxinβ_receptor(LTβR)signaling.Byusingchimericmiceanddifferentvitamin
Adiets,wedescribehowRAandtheLTpathwayarebothinvolvedinthehomeostasisandthepositioningofsplenic
DCs.Moreover,weshowthattheadditionorthedeprivationofRAdonotaffectthehomeostasisofsplenicDCsin
LTβR-/-chimericmice,ascomparedtoLTβR+/+chimericmice,suggestingthat thepresenceofLTβRsignaling is
requiredtomediatetheeffectsofRAonsplenicDC.Finally,wereportthatvitaminAaffectsalsotheaccumulation
ofcollagenintheMZBC,amatrixproteinwhichcouldbeinvolvedintheaccumulationofCD11b+CD8-ESAM+DCs
in this specific region.Overall, these findings provide a further understandingof themechanisms regulating the
homeostasisofsplenicDCsinordertoprovideafirstlineofdefenseagainstblood-bornepathogens.
|Speaker
*Correspondence:[email protected]
13
Non-cell-autonomoustumorsuppressoractivityoftheCdx2homeoboxgene
inthegutthroughthemicroenvironment
CamilleBalbinot,1|OlivierArmant,2ElisabethMartin,1Jean-NoelFreund,1andIsabelleDuluc1*
1InsermUMR_S1113,Strasbourg,France
2KIT,Karlsruhe,Germany
Abstract
ThehomeotictranscriptionfactorCdx2isamajorregulatorofthehomeostasisoftheconstantly-renewingintestinal
epithelium (Stringer et al, 2012). Human colon cancers with a strong reduction of Cdx2 expression are of bad
prognosis.Here,weanalyzedthepathologicalrelevanceofthelossoffunctionofCdx2inthegut.Mosaic lossof
functionofCdx2inthemouseintestineinducedtheoutgrowthofincompletegastric-typemetaplasiasatthelevelof
thecaecum.Thesemetaplasiasarenon-cancerousas theydidnotspontaneouslyprogress incancer,even inold
animals.However, inapremalignantcontext, they indirectly facilitatedthemalignanttransformationofadjacent
Cdx2-intactpremalignantepithelialcells,leadingtoadenocarcinomadevelopmentfromthesurfaceofthelesionsin
atop-downprocess.Thisindirecteffectislinkedtoprofoundmodificationsofthemicroenvironment,includingthe
increased expression of extracellular matrix components, of cytokines and the infiltration of the stroma by
macrophagesandThandTreglymphocytes.ThedatahighlightanovelandoriginalactivityoftheCdx2homeobox
geneinthegut:itsnon-cell-autonomoustumorsuppressoractivity.
Reference
StringerEJ,DulucI,SaandiT,DavidsonI,BialeckaM,SatoT,BarkerN,CleversH,PritchardCA,WintonDJ,WrightNA,FreundJN,DeschampsJ,BeckF.(2012).Cdx2determinesthefateofpostnatalintestinalendoderm.Development139(3):465-74
|Speaker
*Correspondence:[email protected]
14
RoleofBTLAinhumansystemiclupuserythematosus
MatthieuSawaf1|,RenaudFelten
1,2,Jean-DanielFauny
1,Jacques-EricGottenberg
1,2,HélèneDumortier
1andFanny
Monneaux1
1ImmunopathologieetChimieThérapeutique,UPR3572,CNRS,IBMC(InstitutdeBiologieMoléculaireetCellulaire),UniversitédeStrasbourg,Strasbourg,France.
2ServicedeRhumatologie,HôpitalUniversitairedeHautepierre,Strasbourg,France
Abstract
Thebalancebetweenco-stimulatoryandco-inhibitoryreceptorsdeterminesthefateofimmuneresponses.
Co-inhibitoryreceptorssuchasCTLA-4,PD-1andBTLAcanlimitT-cellactivationandmaydirecttheimmuneresponse
towardtolerance,andthusplayanimportantroleforthepreventionofautoimmunity.Recently,inhibitoryreceptors
havedrawnmuchattentionaspotentialtargetsforimmunotherapiesinautoimmunediseases.Moreover,promising
results obtainedwith the use of Abatacept (CTLA4-Ig) in various autoimmune diseases, highlight the interest of
targeting such receptors. Themaingoalof thisproject is tounderstand the involvementof thenewlydescribed
coinhibitoryreceptorBTLA(BandTlymphocyteattenuator)insystemiclupuserythematosus(SLE).Thespecificaims
ofthisprojectaretoanalyzethecellsurfaceexpressionofBTLAonhumanlymphocytesubsetsandtoevaluateBTLA
regulationofTandBcellactivationinlupussettings.
|Speaker
*Correspondence:[email protected]
15
RegulationoflymphnodemacrophagedifferentiationbyRANKL
MélanieChypre1,2|,OlgaCordeiro1,FaroukAlloush1,TobyLawrence3,ChristopherG.Mueller1*
1CNRSUPR3572,IBMC,UniversityofStrasbourg;2PrestwickChemical,Illkirch;3CNRS-INSERM,CIML,Aix-MarseilleUniversity,France
Abstract
The TNF superfamily member RANKL functions in osteoclastogenesis by activating RANK in myeloid osteoclast
precursors.However,whetheritalsoplaysaroleinthedifferentiationofothermacrophagesubsetsisnotknown.
We addressed this question by conditionally deleting RANKL from marginal reticular stromal cells (MRCs) that
constitutively express RANKL in the lymph node.Weobserved impaired differentiation of the subcapsular sinus
macrophages (SSMs) leading to dysfunctional antigen transport to B cells and viral infection. To understand the
mechanisms behind SSMs regulation by RANKL, we generated mice with conditional deficiency of RANK in
macrophagesandMRCs.However,theSSMpopulationwasnormal,excludingdirecteffectofRANKLonlymphnode
macrophagesandMRCs.WehaverecentlyshownthatRANKLactivatedRANK[1].WewereabletorecoverITGA2B
expression on LEC after recombinant RANKL administration to mice deficient in stromal RANKL. We are now
investigatingifthereisacross-talkbetweenRANKL-activatedLECsandSSMs.+
lymphaticendothelialcells(LECs)to
expressITGA2b
Reference
CordeiroOG,ChypreM,BrouardN,RauberS,AlloushF,Romera-HernandezM,etal.Integrin-AlphaIIbIdentifiesMurineLymphNodeLymphaticEndothelialCellsResponsivetoRANKL.PLOSONE.2016;11:e0151848.doi:10.1371/journal.pone.0151848
|Speaker
*Correspondence:[email protected]
16
RoleofTrib1inBcells
SimoniL.1,DelgadoV.
1|,Ruer-LaventieJ.
1,SoleyA.
1,2,DuvalM.
1,PasqualiJL.
1,2,MartinT.
1,2,KorganowAS.
1,2,Soulas-
SprauelP1,2.
1ICT,UPR3572,IBMC,Strasbourg;2HôpitauxUniversitairedeStrasbourg,NouvelHôpitalCivil,Strasbourg
Abstract
TRIB1isapseudokinasethatbelongstothetribblesproteinsfamilyandisimplicatedinvarioushumandiseases.In
thisprojectweshowhowTrib1isinvolvedininBcellfunction,studyingitseffectonhuman,miceandthemurineB
celllineCH12F3.Agenome-widetranscriptomeanalysisofCD19+Bcellscarriedouton10healthycontrolsand17
SystemicLupusErythematosus (SLE)patients in remissionphasehas revealed thatTRIB1 isoverexpressed inSLE
patients.WeproducedmiceoverexpressingTrib1inBcellsfromapro-Bstage:theyhaveadefectinthesecretionof
immunoglobulins,withasignificantdecreaseofserumIgG1productionattheageof6months,aswellasinthetotal
IgGandalsoIgG1afterstimulationoftotalsolenocyteswithLPS+IL-4invitro.InCH12F3Bcellstheoverexpression
ofTrib1inducesareductionintheproductionofIgAafterstimulationwithα-CD40,TGF-βandIL-4.Inthiscellular
modelwehaveanalyzedbymassspectrometrythepartnersofTrib1,identifyingCOP1andCD72astwoofthemains
partners.TheseresultshavebeenvalidatedbyimmunoprecipitationfollowedbyWestern-Blot.Nowadays,weare
developing functional assays to verify the roleof Trib1 in thedefectof immunoglobulinproduction through the
bindingtothesetwopartners.
| Speaker
17
Studyoftype2immunityinatopicdermatitis
RuichengWei|1
1DepartmentofFunctionalGenomicandCancer,IGBMC,Illkirch
Abstract
Atopicdermatitis(AD)isoneofthemostcommoninflammatoryskindiseases,characterizedbychroniccutaneous
inflammation,hyperIgEandThelpertype2(Th2)response.TheprevalenceofADhasincreased,withthenumber
of children suffering fromAD tripled in industrialized nations in the past 30 years, nowaffecting 15-30%of the
childrenand2-10%ofadult.Moreover,ADoftenprogressestootheratopicdiseasessuchasasthma.Ithasbeen
thusurgedtoachieveabetterunderstandingofADandtodevelopeffectivepreventionandtreatmentstrategies.
Ithasbeenrecognizedthattype2immuneresponseiscriticallyimplicatedinthepathogenesisofAD.Usingmouse
modelsandgenetictools,thepreviousstudiesinmylabhaveestablishedacentralroleofcytokinethymicstromal
lymphopoietin(TSLP),whichisexpressedbyepidermalkeratinocytes,inpromotingTh2cellularresponseanddriving
thepathogenesisAD.HereIwilltalkaboutourrecentexplorationonthedifferentiationandregulationofTfollicular
helpercell,anewCD4helperTcellsubsetthathasemergedtobeacriticalplayerinhumoralimmunity,duringAD
pathogenesis.
| Speaker
18
P140peptide,aheatshockproteininhibitor,targetslysosomes
forlupustherapy
FengjuanWang,1| InmaculadaTasset,2AnaMariaCuervo,2andSylvianeMuller1,3*
1CNRS,Immunopathologyandtherapeuticchemistry/LaboratoryofexcellenceMEDALIS,InstitutdeBiologieMoléculaireetCellulaire,
Strasbourg,France;2Departmentofdevelopmentalandmolecularbiology,EinsteinCancerCenter,AlbertEinsteinCollegeofmedicine,
Bronx,NY,USA;3UniversityofStrasbourgInstituteforAdvancedStudy,Strasbourg,France
Abstract
Lysosomesplaypivotalrolesinimmunecellfunctionsandlysosomaldysfunctionhasbeenproposedtocontribute
to the mechanisms of systemic lupus erythematosus, a multifactorial disease characterized by autoimmune
responses where massive amount of autoantibodies target self tissues and organs. Targeting lysosomes has
thereforebeenconsideredasapromisingtherapeuticstrategyforlupus(WangandMuller,2015).P140/LupuzorTM
isaphosphorylated21-merpeptidewhichexhibitssignificantprotectivepropertiesinpatientsandmicemodels
withlupus.ItiscurrentlyunderphaseIIIclinicaltrialinUS,EuropeandcountriesoftheWestIndianOcean.We
havepreviouslydemonstratedthatP140bindsandinhibitsHSPA8/HSC70heatshockprotein(Pageetal.,2011;
Macrietal.,2015).WenowdemonstratethatP140accumulatestolysosomesandmodulateschaperonemediated
autophagy(CMA),aselectivelysosomaldegradativepathway,whichisderegulatedinlupus.Wepostulatethat
theinhibitioneffectofP140onCMAisrelatedtoitshamperingeffectonHSPA8,whichisakeycomponentinCMA.
Reference
WangandMuller(2015)Manipulatingautophagicprocessesinautoimmunediseases:aspecialfocusonmodulatingchaperone-mediatedautophagy,anemergingtherapeutictarget.FrontImmunol.6:252.
Pageetal.(2011)HSC70blockadebythetherapeuticpeptideP140affectsautophagicprocessesandendogenousMHCIIpresentationinmurinelupus.AnnRheumDis,70:837–843.
Macrietal.(2015)Modulationofderegulatedchaperone-mediatedautophagybyaphosphopeptide.Autophagy,11:472-86.
| Speaker*Correspondence:[email protected]
19
P140asamodulatorofautophagyinSjögren'ssyndrome
BaihuiLi1|andSylvianeMuller1,2*
1CNRS,Immunologyandtherapeuticchemistry/LaboratoryofexcellenceMEDALIS,InstitutdeBiologieMoléculaireetCellulaire(IBMC),Strasbourg;2UniversityofStrasbourgInstituteforAdvancedStudy,Strasbourg
Abstract
Sjögren'ssyndrome(SjS)isasystemicautoimmunediseasethataffectsexocrineglandsandcausesinflammation
and dysfunction in glandular tissues. Salivary glands and lacrimal glands are predominantly affected, leading to
symptomsofdryeyesanddrymouthrespectivelyinSjS.HereweuseMRL/lprmousemodeltostudyautophagy,a
conserveddegradationpathway,insalivaryglandsofthesemice.Weinducedautophagybystarvationofsalivary
glandcellsandmeasuredautophagicmarkerssuchasMAP1LC3B-IIandSQSTM1/p62.Ourresultsshowedthatthere
isnoautophagic flux in starvedsalivaryglandcellsofMRL/lprmice,while in cellsof thecontrolgroup (healthy
C57BL/6mice),thereisanactivefluxwithaccumulationofMAP1LC3B-IIandSQSTM1/p62proteinsafterautophagy
induction. This suggests a deficiency of autophagy in the SjS setting in MRL/lpr mice. Upon intravenous
administration of P140 peptide, MRL/lpr mice restored autophagy and partially ameliorated SjS-related
extraglandularmanifestations.TheseresultssuggestthatautophagyisinvolvedinthepathogenesisofSjSandthat
P140couldbeapromisingtreatmentforSjS.
Reference:Esteban-Martínez,L.andP.Boya,Autophagicfluxdeterminationinvivoandexvivo.Methods,2015.75:79-86.
Macri,C.,etal.,Modulationofderegulatedchaperone-mediatedautophagybyaphosphopeptide.Autophagy,2015.
11(3):472-86.
|Speaker*Correspondence:[email protected]
20
MechanismsofactionofanCXCL12inhibitorintheanti-inflammationofasthma
FDaubeuf1|,DBonnet1,VBeckaert3,AOuadi3,PMarchand3,DBrasse3,PGizzi2,MHibert1,JLGalzi2,NFrossard1*
1UMR7200;2UMR7242CNRS-UDS,ILLKIRCH;3Imabio,IPHC,STRASBOURG,France
AbstractTheCXCL12chemokineanditsreceptorsCXCR4-CXCR7are involvedinnormaltissuepatterning,aswellas inthe
physiopathologyof inflammatorydiseases, includingallergicasthma.Werecently reported inamurinemodelof
airway hypereosinophilia the anti-inflammatory action of a non-peptidic CXCL12 neutraligand, chalcone 4. We
synthesized an 123I-chalcone 4, exhibiting similar activities to chalcone 4, to visualize its bioavailability when
administeredintranasally.Weshowrapideliminationof123I-chalcone4fromthelunginlessthan30min(95±5%),
andeliminationinthebile,fecesandurine.PKanalysisconfirmstheeliminationhalf-life(T½<5min)inthelung.This
isconcomitanttoadecreaseinCXCL12inlung(-20%),anditsincreaseinblood(x1000),suggestingdrainingofthe
neutraligand and CXCL12 from the lung. Although chalcone 4 is eliminated, we report the inhibition of airway
hyperresponsiveness(-45%),inflammatorycellrecruitment,inparticulareosinophils(-54%)andM1macrophages(-
65%), and remodeling, mucus hypersecretion (-84%) and collagen deposition (-78%). In addition, we show an
inhibitionofthealveolarmacrophageM1-M2polarizationaccompaniedbyinhibitionofcytokinereleaseexvivoin
response to ovalbumin (TNF-α: -99%, IL-5: -91%) or to CXCL12 itself (TNF-α: -98%), suggesting a role of the
macrophageinmodulatingtherecruitmentofeosinophils.Inconclusion,theCXCL12neutraligandbindsendogenous
CXCL12 to prevent binding to its receptor and glycosaminoglycans, thereby collapsing the extracellular density
gradient of CXCL12 to escape the immune response, and inhibits macrophage activation and differentiation in
responsetoallergen.
|Speaker*Correspondence:[email protected]
21
DevelopmentofanovelimmunotherapywithTLR/NLRsynergyfor
targetedcancertherapy
CéliaJacoberger-Foissac,1| ,BenoitFrisch1BéatriceHeurtault,1*SylvieFournel,1*
1EquipedeBiovectorologie,LaboratoiredeConceptionetApplicationdeMoléculesBioactives,UMR7199CNRS/Unistra,Facultéde
Pharmacie,Illkirch.
Abstract
Currently, a challenging goal in the area of cancer treatment is the development of innovative targeted
antitumoralimmunotherapieswithalong-termefficiency.Inthiscontext,myteamtookadvantageofliposomal
nanoparticlespropertiesfortheconceptionofcancervaccines.Inapreviousstudy,thecombinationonliposomal
surfaceof threeelements(twopeptideepitopes includingonethat isspecifictotumorcellsandanadjuvant,
ligandofaToll-likereceptor)crucialforimmuneresponsehasdemonstratedtoinducecompleteregressionof
tumor growth after prophylactic or therapeutic treatments in mice grafted with a tumor [1]. However, the
therapeutictreatmentefficiencyquicklydecreasedwiththeincreaseofthetimespentbetweentumorgrafting
andtreatmentstart[2].
Inordertooptimizeourtreatmentandshowtheuniversalityofournanoparticleapproach,weproposedto
validateourstrategyinanothertumormousemodelandtotryforthefirsttimeacombinationofadjuvants(ligand
ofTLR2/6andNOD1). The combinationof these twomolecules ina liposome formulation containingpeptide
epitopesallowedusto increasethetherapeuticwindowofourtreatment. Indeed,weobservedadecrease in
tumorgrowthof60% incomparisontobothadjuvantsalone (5%and40%respectively), if the liposomesare
injectedondays8and10afterthetumorimplantation.
References[1]ThomannJ.S.,etal.Biomaterials2011,32:4574-4583[2]RothA.,etal.BritishJournalofCancer2005,92:1421-1429
| Speaker*Correspondence:[email protected]/[email protected]
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