Generation of CD8+ and CD4+ Regulatory T Cells from Human Pluripotent Stem Cells

Wednesday, June 19

2:05 PM - 2:20 PM

Location: Salon H-K, 4th Floor

Presenting Author(s)

LF

Lea Flippe

Ph.D Student
Centre de Recherche en Transplantation et Immunologie UMR 1064, Inserm, Nantes, France; Institut de Transplantation Urologie Néphrologie, Nantes, France

Co-Author(s)

AG

Anne Gaignerie

iPSC corefacility, University of Nantes; INSERM UMS 016; CNRS 3556; CHU de Nantes, France
SB

Séverine Bézie

Centre de Recherche en Transplantation et Immunologie UMR 1064, Inserm, Nantes, France; Institut de Transplantation Urologie Néphrologie, Nantes, France; LabEx IGO "Immunotherapy, Graft, Oncology", Nantes, France.
IA

Ignacio Anegon

Centre de Recherche en Transplantation et Immunologie UMR 1064, Inserm, Nantes, France; Institut de Transplantation Urologie Néphrologie, Nantes, France; LabEx IGO "Immunotherapy, Graft, Oncology", Nantes, France.
LD

Laurent David

Centre de Recherche en Transplantation et Immunologie UMR 1064, Inserm, Nantes, France; Institut de Transplantation Urologie Néphrologie, Nantes, France, iPSC corefacility, University of Nantes; INSERM UMS 016; CNRS 3556; CHU de Nantes, France
Carole Guillonneau

Centre de Recherche en Transplantation et Immunologie UMR 1064, Inserm, Nantes, France; Institut de Transplantation Urologie Néphrologie, Nantes, France; LabEx IGO

Organ or cell transplantation is the only therapeutic solution for pathologies causing an irreversible loss of vital organs function. The development of novel specific and non-toxic anti-rejection immunotherapies is a major goal in transplantation. Strategies based on regulatory T cells (Tregs) are promising. However, Tregs cell-based therapies have been hampered by the technical limitation of obtaining large batches of functional Tregs. The project aim is to obtain an unlimited number of Tregs from human pluripotent stem cells (hPSCs). We have developed a new differentiation protocol in two separate steps: the first step is to generate hematopoietic stem cells (HSCs) from hPSCs and the second is to engage HSCs towards the lymphoid lineage. We have generated HSCs thought embryoid bodies (EBs) formation for 9 days. After 9 days, we obtained 50% of cells expressing CD34⁺, a key marker of HSCs. Then, we dissociated EBs and co-cultured them onto OP9-DLL1 to induce T-lymphoid differentiation for 26 days. At day 10 of co-culture, we transduced our cells with a lentivirus encoding FOXP3, a known master transcription factor of Tregs. FOXP3 transduction at day 10 of co-culture resulted in significant differentiation of Foxp3⁺CD3⁺TCRαβ⁺CD8⁺ or CD4⁺Tregs cells until day 26. To our knowledge, it’s the first time that CD4+ and CD8+ Tregs have been differentiated from hPSCs. Moreover, our result support that human CD4⁺ and CD8⁺Tregs differentiation would be under the control of Foxp3. Our findings open new possibilities for cell therapy.