title: Only the CD45RA+ subpopulation of CD4+CD25high T cells gives rise to homogeneous regulatory T-cell lines upon in vitro expansion.
abstract: Thymus-derived CD4+ CD25+ regulatory T cells suppress autoreactive CD4+ and CD8+ T cells and thereby protect from autoimmunity. In animal models, adoptive transfer of CD4+ CD25+ regulatory T cells has been shown to prevent and even cure autoimmune diseases as well as pathogenic alloresponses after solid organ and stem-cell transplantations. We recently described methods for the efficient in vitro expansion of human regulatory T cells for clinical applications. We now demonstrate that only CCR7- and L-selectin (CD62L)-coexpressing cells within expanded CD4+ CD25high T cells maintain phenotypic and functional characteristics of regulatory T cells. Further analysis revealed that these cells originate from CD45RA+ naive cells within the CD4+ CD25high T-cell compartment, as only this subpopulation homogeneously expressed CD62L, CCR7, cytotoxic T lymphocyte-associated antigen-4 (CTLA-4), and forkhead box P3 (FOXP3), produced no inflammatory cytokines and maintained robust suppressive activity after expansion. In contrast, cell lines derived from CD45RA- memory-type CD4+ CD25high T cells lost expression of lymph node homing receptors CCR7 and CD62L, contained interleukin-2 (IL-2) and interferon-gamma (IFN-gamma) as well as IL-10-secreting cells, showed only moderate suppression and, most importantly, did not maintain FOXP3 expression. Based on these unexpected findings, we suggest that isolation and expansion of CD45RA+ naive CD4+ CD25high T cells is the best strategy for adoptive regulatory T (Treg)-cell therapies.
authors.affiliation: Institute of Immunology, Department of Hematology & Oncology, University Hospital Regensburg, Regensburg, Germany. petra.hoffmann@klinik.uni-regensburg.de
authors.forename: Petra
authors.initials: P
authors.lastname: Hoffmann
authors.forename: Ruediger
authors.initials: R
authors.lastname: Eder
authors.forename: Tina J
authors.initials: TJ
authors.lastname: Boeld
authors.forename: Kristina
authors.initials: K
authors.lastname: Doser
authors.forename: Biserka
authors.initials: B
authors.lastname: Piseshka
authors.forename: Reinhard
authors.initials: R
authors.lastname: Andreesen
authors.forename: Matthias
authors.initials: M
authors.lastname: Edinger
mesh_terms: D015703:Antigens, CD / Q000096:biosynthesis / Q000276:immunology; D000943:Antigens, Differentiation / Q000096:biosynthesis / Q000276:immunology; D060908:CTLA-4 Antigen; D002454:Cell Differentiation / Q000276:immunology*; D049109:Cell Proliferation; D002478:Cells, Cultured; D016207:Cytokines / Q000096:biosynthesis / Q000276:immunology; D005260:Female; D051858:Forkhead Transcription Factors / Q000096:biosynthesis / Q000276:immunology; D005786:Gene Expression Regulation / Q000276:immunology*; D006801:Humans; D016219:Immunotherapy, Adoptive; D019041:L-Selectin / Q000096:biosynthesis / Q000276:immunology; D017493:Leukocyte Common Antigens / Q000096:biosynthesis / Q000276:immunology*; D008297:Male; D054400:Receptors, CCR7; D019707:Receptors, Chemokine / Q000096:biosynthesis / Q000276:immunology; D050378:T-Lymphocytes, Regulatory / Q000166:cytology / Q000276:immunology* / Q000378:metabolism; D013950:Thymus Gland / Q000166:cytology / Q000276:immunology* / Q000378:metabolism
chemical_list: D015703:Antigens, CD; D000943:Antigens, Differentiation; C515370:CCR7 protein, human; D060908:CTLA-4 Antigen; C556706:CTLA4 protein, human; D016207:Cytokines; C418974:FOXP3 protein, human; D051858:Forkhead Transcription Factors; D054400:Receptors, CCR7; D019707:Receptors, Chemokine; D019041:L-Selectin; D017493:Leukocyte Common Antigens