Human autologous tolerogenic dendritic cells impair T-cell proliferation through contact-independent mechanisms

(1)

HAL Id: inserm-02160928

https://www.hal.inserm.fr/inserm-02160928

Submitted on 20 Jun 2019

HAL is a multi-disciplinary open access

archive for the deposit and dissemination of sci-entific research documents, whether they are pub-lished or not. The documents may come from teaching and research institutions in France or abroad, or from public or private research centers.

L’archive ouverte pluridisciplinaire HAL, est destinée au dépôt et à la diffusion de documents scientifiques de niveau recherche, publiés ou non, émanant des établissements d’enseignement et de recherche français ou étrangers, des laboratoires publics ou privés.

Human autologous tolerogenic dendritic cells impair T-cell proliferation through contact-independent

mechanisms

Eros Marín, Laurence Bouchet-Delbos, Maria-Cristina Cuturi, Aurelie Moreau

To cite this version:

Eros Marín, Laurence Bouchet-Delbos, Maria-Cristina Cuturi, Aurelie Moreau. Human autologous tolerogenic dendritic cells impair T-cell proliferation through contact-independent mechanisms. 22nd NAT congress ”Immunotherapies in transplantation and Cancer”, Jun 2017, Nantes, France. �inserm-02160928�

(2)

Treg Thxkx0 KTDC CDE( KllogeneicymDC Solubleyfactors _{Solubleyfactors} TGFv CDE(mDC CDE(mDC CDE(mDC(KTDC CDE(mDC CDE(mDC(KTDC KTDC Monocytey elutriation KIMV GMvCSF RPMI GMvCSF ILvE Hydays DonoryK DonoryB

y

HumanGautologousGtolerogenicGdendriticGcellsGimpairGT2cellG

proliferationGthroughGcontact2independentGmechanisms

ATDCGimpairGCD4

)

GT2cellGproliferationGthroughG

contactGindependentGmechanisms

Background

ATDCGimpairGTh1GandGTh17GproliferationGwhileG

promoteGTreg

ATDCGimpairGCD4)T2cellGproliferationGthroughGG0/G1G

phaseGblockadeGwithoutGinducingGapoptosisGnorGanergyG

ATDCGinduceGtheGgenerationGofG/deGnovo/GTregG

partiallyGthroughGtheGproductionGofGTGF2G

Conclusion

Model

Counts Suppressionyassay Transwellyassay

y

Kutologousy tolerogenicy dendriticy cellsy BKTDCpy havey beeny demonstratedy toy bey abley toy increasey heart)yskinyandypancreaticyisletyallograftysurvivalsyinymurineymodelsyandytoybeysafeyinynonvhumany primates,yDueytoytheiryefficacyyandytheirysafety)yKTDCytherapyyhasybeenymovedytoyclinicyinyayfirsty phasey IkIIy clinicaly trialy iny kidneyy transplantationy iny they contexty ofy they ONEy studyy project,y Humany KTDCyareygeneratedyfromyelutriatedymonocytesyculturedyHydaysyinyKIMVymediumysupplementedy withy lowy dosey ofy GMvCSF,y Kftery differentiationy KTDCy displayy ay tolerogenicy dendriticy celly profiley Bi,e,y theyy displayy any immaturey phenotype)y theyy expressy immunomodulatoryy cytokinesy suchy asy ILvx.yandyTGFvyy)ytheyyareyresistantytoymaturationyandytheyyhaveysuppressiveyactivityytowardyTv cellsp,y Thisy projecty aimsy toy determiney they suppressivey mechanismsy ofy humany KTDCy towardy Tv cells,

y

xyCentreydeyRechercheyenyTransplantationyetyImmunologieyUMRx.HE)yINSERM)yUniversitéydeyNantes)yNantes)yFrance

OyInstitutydeyTransplantationyUrologieyNéphrologieyBITUNp)yCHUyNantes)yNantes)yFrance

ErosyMarin

x)O

)yLaurenceyBouchetvDelbos

x)O

)yMariayCristinayCuturi

x)O

)yKurélieyMoreau

x)O

y

FigureG1.GMA.yCDE(yTvcellyproliferationyinysuppressionyassayyandyMB.ytranswellyassay

FigureG3.GBA.GPercentageyofyproliferatingyCDE(tvbet(IFNvyy(yThxycells)y MB.yCDE(ILx0K(yThx0ycells)yandyMC.yCDE(CDOjhiFoxPNhiyTregycellsyy

afterysuppressionyassay

FigureG 4.G MAG andG B.G Suppressiony assayy startingy withy CDE(CDOjvy Tvcellsy

insteady ofy totaly CDE(y Tvcells,G MA.y Percentagey ofy proliferatingy CDE (CDOjhiFoxPNhiyTregycellsyandyMB.yafteryaddingyTGFvyyyblockingyantibody,y

FigureG2.GMA.yCDE(Tvcellycellycycleyandyy MB.G apoptosisy iny suppressiony assay,y G G MC.y Knergyy assay,y First)y CDE(y Tvcellsy

werey culturedy withy allogeneicy KTDCy ory MoDC,y Kftery Hyday)yCDE(yTvcellsywerey recoveredy andy culturedy withy allogeneicy mDC,

Thesey resultsy altogethery showedy thaty lowy dosey GMvCSFvinducedy KTDCy arey ableytoysuppressyCDE(TvcellyproliferationyandytoyinduceyVdeynovoVyTregyinyparty throughytheyexpressionyofyTGFvyy,yMoreoveryweyevincedythatyKTDCyimpairyTv celly proliferationy byy contactvindependenty mechanismsy withouty assotiationy withy apoptosisy nory anergyy buty withy they blockadey ofy celly cycley iny G.kGxy phase,y Inv depthyinvestigationsywillybeyperformedytoydetermineytheymolecularymechanismsy underlyingyKTDC, FigureG1.GBA.ySchematicyrepresentationyofyKTDCyandyMoDCydifferentiation,yInythisy studyyweyusedyMoDCyasycontrolycells,yMB.yModelyofysuppressionyassayyusedyinythisy study,yCDE(TvcellsyareyculturedywithyallogeneicymatureydendriticycellsyBmDCpywithy orywithoutyKTDCyoryMoDCyBautologousytoyCDE(yTvcellsp KTDC Tvcells Or LPS CDE( mDC y MoDC KTDC v v v ( ( v v ( ( v ( ( ( ( v ( j)0O _N)H. O)éé U0)0 x.. x.x x.O x.N x.E x.j x.. x.x x.O x.N x.E x.j N)xx O)éU O)é0 éx). x.. x.x x.O x.N x.E x.j x.. x.x x.O x.N x.E x.j Gx_G. 0U)N GO_M é)0E S xx)j . j.K x..K xj.K O..K Oj.K . j.. x).K x)jK O).K Gx_G. éO)x GO_M O)00 S E)0U . j.K x..K xj.K O..K Oj.K . x).K O).K N).K CDE( mDC KTDC v v ( ( v ( ( ( ( mDC CDE(BKTDCp ( v v ₍ ₍ CDE(BMoDCp v ( v ₍ v ₍ v KnnexinvV DKPI PropidiumyIodide CDE(Tvcellycellycycleyafterysuppressionyassay CDE(Tvcellyapoptosisyafterysuppressionyassay Knergyyassay CDE(mDC CDE(mDC(KTDC CDE(mDC CDE(mDC(KTDC VDeynovoVyTregyinduction VDeynovoVyTregyinductionyiny presenceyofyantivTGFv

(

v

CDE(mDC(KTDC Hx)x O.)0 N N E j . vx.N x.N x.E x.j . vx. x. x. x. E)xj N N E j . vx.N x.N x.E x.j x)xH . vx.N x.N x.E x.j . vx.N x.N x.E x.j N)Hj . vx.N x.N x.E x.j . vx.N x.N x.E x.j xU). . vx.N x.N x.E x.j . vx.N x.N x.E x.j Nj)E Ej)x N N E j . vx.N x.N x.E x.j IFNv tvbet ILvx0K CDxHx CDOj FoxPN