title: Freeze and Thaw of CD4+CD25+Foxp3+ Regulatory T Cells Results in Loss of CD62L Expression and a Reduced Capacity to Protect against Graft-versus-Host Disease.
abstract: The adoptive transfer of CD4+CD25+Foxp3+ regulatory T cells (Tregs) in murine models of allogeneic hematopoietic cell transplantation (HCT) has been shown to protect recipient mice from lethal acute graft-versus-host disease (GVHD) and this approach is being actively investigated in human clinical trials. Here, we examined the effects of cryopreservation on Tregs. We found that freeze and thaw of murine and human Tregs is associated with reduced expression of L-selectin (CD62L), which was previously established to be an important factor that contributes to the in vivo protective effects of Tregs. Frozen and thawed murine Tregs showed a reduced capacity to bind to the CD62L binding partner MADCAM1 in vitro as well as an impaired homing to secondary lymphoid organs in vivo. Upon adoptive transfer frozen and thawed Tregs failed to protect against lethal GVHD compared with fresh Tregs in a murine model of allogeneic HCT across major histocompatibility barriers. In summary, the direct administration of adoptively transferred frozen and thawed Tregs adversely affects their immunosuppressive potential which is an important factor to consider in the clinical implementation of Treg immunotherapies.
authors.affiliation: Division of Blood and Marrow Transplantation, Department of Medicine, Stanford University School of Medicine, Stanford, CA, United States of America.
authors.forename: Mareike
authors.initials: M
authors.lastname: Florek
authors.affiliation: Division of Blood and Marrow Transplantation, Department of Medicine, Stanford University School of Medicine, Stanford, CA, United States of America.|Department of Medicine II, Eberhard Karls University, Tübingen, Germany.
authors.forename: Dominik
authors.initials: D
authors.lastname: Schneidawind
authors.affiliation: Division of Blood and Marrow Transplantation, Department of Medicine, Stanford University School of Medicine, Stanford, CA, United States of America.|Hematology and Clinical Immunology, Department of Medicine, University of Perugia, Perugia, Italy.
authors.forename: Antonio
authors.initials: A
authors.lastname: Pierini
authors.affiliation: Division of Blood and Marrow Transplantation, Department of Medicine, Stanford University School of Medicine, Stanford, CA, United States of America.
authors.forename: Jeanette
authors.initials: J
authors.lastname: Baker
authors.affiliation: Division of Blood and Marrow Transplantation, Department of Medicine, Stanford University School of Medicine, Stanford, CA, United States of America.
authors.forename: Randall
authors.initials: R
authors.lastname: Armstrong
authors.affiliation: Division of Blood and Marrow Transplantation, Department of Medicine, Stanford University School of Medicine, Stanford, CA, United States of America.
authors.forename: Yuqiong
authors.initials: Y
authors.lastname: Pan
authors.affiliation: Division of Blood and Marrow Transplantation, Department of Medicine, Stanford University School of Medicine, Stanford, CA, United States of America.
authors.forename: Dennis
authors.initials: D
authors.lastname: Leveson-Gower
authors.affiliation: Division of Blood and Marrow Transplantation, Department of Medicine, Stanford University School of Medicine, Stanford, CA, United States of America.
authors.forename: Robert
authors.initials: R
authors.lastname: Negrin
authors.affiliation: Division of Blood and Marrow Transplantation, Department of Medicine, Stanford University School of Medicine, Stanford, CA, United States of America.
authors.forename: Everett
authors.initials: E
authors.lastname: Meyer
mesh_terms: D019264:Adoptive Transfer; D064591:Allografts; D000818:Animals; D015815:Cell Adhesion Molecules / Q000276:immunology; D015925:Cryopreservation; D004195:Disease Models, Animal; D005786:Gene Expression Regulation / Q000276:immunology*; D006086:Graft vs Host Disease / Q000276:immunology* / Q000473:pathology / Q000517:prevention & control; D018380:Hematopoietic Stem Cell Transplantation; D006801:Humans; D007136:Immunoglobulins / Q000235:genetics / Q000276:immunology; D019041:L-Selectin / Q000276:immunology*; D051379:Mice; D008807:Mice, Inbred BALB C; D009088:Mucoproteins / Q000235:genetics / Q000276:immunology; D050378:T-Lymphocytes, Regulatory / Q000276:immunology* / Q000473:pathology / Q000637:transplantation
chemical_list: D015815:Cell Adhesion Molecules; D007136:Immunoglobulins; C504265:MADCAM1 protein, human; C489082:Madcam1 protein, mouse; D009088:Mucoproteins; D019041:L-Selectin