title: Adoptive transfer of allogeneic regulatory T cells into patients with chronic graft-versus-host disease.
abstract: OBJECTIVE
  Mouse models indicate that adoptive transfer of regulatory T cells (Treg) may suppress graft-versus-host-disease (GvHD) while preserving graft-versus-leukemia reactions. We aimed to develop a protocol for the efficient isolation and in vitro expansion of donor-derived Treg and to establish the proof-of-concept for the clinical application of ex vivo-generated Treg preparations in five patients with otherwise treatment-refractory chronic GvHD (cGvHD).
  
  
  METHODS
  Allogeneic Treg were isolated from unstimulated leukapheresis products of the corresponding human leukocyte antigen-matched donors by use of clinical-grade magnetic-activated bead sorting. To increase the amount and purity, Treg were cultivated for 7-12 days and infused after a median time of 35 months after allogeneic hematopoietic cell transplantation.
  
  
  RESULTS
  Final products contained Treg with a median purity of 84.1% CD4(+)CD25(high)CD127(low)FOXP3(+)of CD45(+) cells and a mean quantity of 2.4 × 10(6) Treg per kg body wt. All isolated cell products showed in vitro suppressive activity. On transfusion, two of five patients showed a clinical response with improvement of cGvHD symptoms. The other three patients showed stable cGvHD symptoms for up to 21 months. In four of five patients, increased counts of Treg were detectable on Treg transfusion, immunosuppressive treatment could be reduced and suppression of CD69 activation marker expression on T-effector cells was observed. However, one patient had development of malignant melanoma and another patient had Bowen skin cancer 4 months and 11 months after Treg transfusion, respectively.
  
  
  CONCLUSIONS
  We demonstrate a feasible and reproducible approach of isolating functional Treg in high quantity and purity for clinical application and show opportunities and risks of adoptive Treg transfer into patients with cGvHD.
authors.affiliation: DFG Center for Regenerative Therapies, Technische Universität Dresden, Dresden, Germany.
authors.forename: Anke
authors.initials: A
authors.lastname: Theil
authors.affiliation: Medical Clinic I, University Hospital Dresden, Technische Universität Dresden, Dresden, Germany.
authors.forename: Sebastian
authors.initials: S
authors.lastname: Tuve
authors.affiliation: Medical Clinic I, University Hospital Dresden, Technische Universität Dresden, Dresden, Germany.
authors.forename: Uta
authors.initials: U
authors.lastname: Oelschlägel
authors.affiliation: Medical Clinic I, University Hospital Dresden, Technische Universität Dresden, Dresden, Germany.
authors.forename: Anja
authors.initials: A
authors.lastname: Maiwald
authors.affiliation: Medical Clinic I, University Hospital Dresden, Technische Universität Dresden, Dresden, Germany.
authors.forename: Diana
authors.initials: D
authors.lastname: Döhler
authors.affiliation: Medical Clinic I, University Hospital Dresden, Technische Universität Dresden, Dresden, Germany.
authors.forename: Dennis
authors.initials: D
authors.lastname: Oßmann
authors.affiliation: Medical Clinic I, University Hospital Dresden, Technische Universität Dresden, Dresden, Germany.
authors.forename: Anja
authors.initials: A
authors.lastname: Zenkel
authors.affiliation: DFG Center for Regenerative Therapies, Technische Universität Dresden, Dresden, Germany.
authors.forename: Carmen
authors.initials: C
authors.lastname: Wilhelm
authors.affiliation: Medical Clinic I, University Hospital Dresden, Technische Universität Dresden, Dresden, Germany.
authors.forename: Jan Moritz
authors.initials: JM
authors.lastname: Middeke
authors.affiliation: Medical Clinic I, University Hospital Dresden, Technische Universität Dresden, Dresden, Germany.
authors.forename: Nona
authors.initials: N
authors.lastname: Shayegi
authors.affiliation: Medical Clinic I, University Hospital Dresden, Technische Universität Dresden, Dresden, Germany.
authors.forename: Karolin
authors.initials: K
authors.lastname: Trautmann-Grill
authors.affiliation: Medical Clinic I, University Hospital Dresden, Technische Universität Dresden, Dresden, Germany.
authors.forename: Malte
authors.initials: M
authors.lastname: von Bonin
authors.affiliation: Medical Clinic I, University Hospital Dresden, Technische Universität Dresden, Dresden, Germany.
authors.forename: Uwe
authors.initials: U
authors.lastname: Platzbecker
authors.affiliation: DFG Center for Regenerative Therapies, Technische Universität Dresden, Dresden, Germany; Medical Clinic I, University Hospital Dresden, Technische Universität Dresden, Dresden, Germany.
authors.forename: Gerhard
authors.initials: G
authors.lastname: Ehninger
authors.affiliation: DFG Center for Regenerative Therapies, Technische Universität Dresden, Dresden, Germany.
authors.forename: Ezio
authors.initials: E
authors.lastname: Bonifacio
authors.affiliation: DFG Center for Regenerative Therapies, Technische Universität Dresden, Dresden, Germany; Medical Clinic I, University Hospital Dresden, Technische Universität Dresden, Dresden, Germany. Electronic address: Martin.Bornhaeuser@uniklinikum-dresden.de.
authors.forename: Martin
authors.initials: M
authors.lastname: Bornhäuser
mesh_terms: D019264:Adoptive Transfer / Q000379:methods*; D000328:Adult; D000818:Animals; D002478:Cells, Cultured; D005260:Female; D006086:Graft vs Host Disease / Q000628:therapy*; D018380:Hematopoietic Stem Cell Transplantation; D006801:Humans; D007165:Immunosuppression Therapy / Q000379:methods*; D007937:Leukapheresis / Q000379:methods; D008297:Male; D051379:Mice; D008875:Middle Aged; D050378:T-Lymphocytes, Regulatory / Q000276:immunology / Q000637:transplantation*; D014019:Tissue Donors; D014184:Transplantation, Homologous
keywords: adoptive cellular therapy; allogeneic hematopoietic cell transplantation; graft-versus-host disease; regulatory T cells