title: Automated isolation of primary antigen-specific T cells from donor lymphocyte concentrates: results of a feasibility exercise.
abstract: BACKGROUND
  The safety and clinical efficacy of adoptive transfer of prospectively isolated antigen-specific T cells are well established. Several competing selection methods are available, one of which is based on immunomagnetic enrichment of T cells secreting IFNγ after incubation with the relevant antigen. The proprietary, GMP-conforming selection technology, called 'cytokine capture system' (CCS) is established in many laboratories for the CliniMACS Plus system. It is robust and efficient, but labour-intensive and incompatible with a single-shift working schedule. An automatic immunomagnetic cell processing system, CliniMACS Prodigy ('Prodigy'), including a protocol for fully automatic CCS execution was recently released.
  
  
  METHODS
  Feasibility of clinical-scale CMV-specific T-cell selection using Prodigy was evaluated using leukoapheresis products from five healthy CMV sero-positive volunteers. Clinical reagents and consumables were used throughout.
  
  
  RESULTS
  The process required no operator input beyond set-up and QC-sample collection, that is, feasibility was given. An IFNγ-secreting target T-cell population was detectable after stimulation, and >2 log-scale relative depletion of not CMV-reactive T cells in the target population was achieved. Purity, that is the frequency of CMV-reactive T cells among all CD3(+) cells ranged between 64 and 93%.
  
  
  CONCLUSIONS
  The CCS protocol on Prodigy is unrestrictedly functional. It runs fully automatically beyond set-up and thus markedly reduces labour. The quality of the products generated is similar to products generated with CliniMACS Plus. The automatic system is thus suitable for routine clinical application.
authors.affiliation: German Red Cross Blood Service Baden-Württemberg-Hesse, Department of Cellular Therapeutics (GMP), Institute Frankfurt, Frankfurt, Germany.
authors.forename: M
authors.initials: M
authors.lastname: Bunos
authors.affiliation: German Red Cross Blood Service Baden-Württemberg-Hesse, Department of Cellular Therapeutics (GMP), Institute Frankfurt, Frankfurt, Germany.
authors.forename: C
authors.initials: C
authors.lastname: Hümmer
authors.affiliation: German Red Cross Blood Service Baden-Württemberg-Hesse, Department of Cellular Therapeutics (GMP), Institute Frankfurt, Frankfurt, Germany.
authors.forename: E
authors.initials: E
authors.lastname: Wingenfeld
authors.affiliation: German Red Cross Blood Service Baden-Württemberg-Hesse, Department of Cellular Therapeutics (GMP), Institute Frankfurt, Frankfurt, Germany.
authors.forename: N
authors.initials: N
authors.lastname: Sorg
authors.affiliation: Center of Child and Adolescent Health, Department for Stem Cell Transplantation and Immunology, Goethe University Medical Center, Frankfurt, Germany.
authors.forename: V
authors.initials: V
authors.lastname: Pfirrmann
authors.affiliation: Center of Child and Adolescent Health, Department for Stem Cell Transplantation and Immunology, Goethe University Medical Center, Frankfurt, Germany.
authors.forename: P
authors.initials: P
authors.lastname: Bader
authors.affiliation: German Red Cross Blood Service Baden-Württemberg-Hesse, Department of Cellular Therapeutics (GMP), Institute Frankfurt, Frankfurt, Germany.|Institute for Transfusion Medicine and Immunohematology, Goethe University Medical Center, Frankfurt, Germany.
authors.forename: E
authors.initials: E
authors.lastname: Seifried
authors.affiliation: German Red Cross Blood Service Baden-Württemberg-Hesse, Department of Cellular Therapeutics (GMP), Institute Frankfurt, Frankfurt, Germany.|Institute for Transfusion Medicine and Immunohematology, Goethe University Medical Center, Frankfurt, Germany.|Department of Medicine, Division of Hematology, University of Washington, Seattle, WA, USA.
authors.forename: H
authors.initials: H
authors.lastname: Bönig
mesh_terms: D016207:Cytokines / Q000235:genetics / Q000378:metabolism; D005434:Flow Cytometry / Q000295:instrumentation / Q000379:methods*; D006801:Humans; D008213:Lymphocyte Activation; D013601:T-Lymphocytes / Q000276:immunology*
chemical_list: D016207:Cytokines
keywords: Adoptive transfer; CCS; CMV; CliniMACS; GMP; automation