title: Isolation of CD4+CD25+ regulatory T cells for clinical trials.
abstract: The adoptive transfer of donor CD4+CD25+ regulatory T cells has been shown to protect from lethal graft-versus-host disease after allogeneic bone marrow transplantation in murine disease models. Efficient isolation strategies that comply with good manufacturing practice (GMP) guidelines are prerequisites for the clinical application of human CD4+CD25+ regulatory T cells. Here we describe the isolation of CD4+CD25+ T cells with regulatory function from standard leukapheresis products by using a 2-step magnetic cell-separation protocol performed under GMP conditions. The generated cell products contained on average 49.5% CD4+CD25high T cells that phenotypically and functionally represented natural CD4+CD25+ regulatory T cells and showed a suppressive activity comparable to that of CD4+CD25+ regulatory T-cell preparations purified by non-GMP-approved fluorescence-activated cell sorting.
authors.affiliation: Department of Hematology and Oncology, University Hospital Regensburg, Regensburg, Germany.
authors.forename: Petra
authors.initials: P
authors.lastname: Hoffmann
authors.forename: Tina J
authors.initials: TJ
authors.lastname: Boeld
authors.forename: Ruediger
authors.initials: R
authors.lastname: Eder
authors.forename: Julia
authors.initials: J
authors.lastname: Albrecht
authors.forename: Kristina
authors.initials: K
authors.lastname: Doser
authors.forename: Biserka
authors.initials: B
authors.lastname: Piseshka
authors.forename: Ashraf
authors.initials: A
authors.lastname: Dada
authors.forename: Claudia
authors.initials: C
authors.lastname: Niemand
authors.forename: Mario
authors.initials: M
authors.lastname: Assenmacher
authors.forename: Evelyn
authors.initials: E
authors.lastname: Orsó
authors.forename: Reinhard
authors.initials: R
authors.lastname: Andreesen
authors.forename: Ernst
authors.initials: E
authors.lastname: Holler
authors.forename: Matthias
authors.initials: M
authors.lastname: Edinger
mesh_terms: D019264:Adoptive Transfer; D000818:Animals; D016026:Bone Marrow Transplantation; D002986:Clinical Trials as Topic; D004195:Disease Models, Animal; D005434:Flow Cytometry / Q000379:methods / Q000592:standards; D006086:Graft vs Host Disease / Q000209:etiology / Q000628:therapy*; D017408:Guidelines as Topic / Q000592:standards; D006801:Humans; D007937:Leukapheresis / Q000379:methods / Q000592:standards; D051379:Mice; D050378:T-Lymphocytes, Regulatory / Q000166:cytology* / Q000637:transplantation; D014184:Transplantation, Homologous