title: Generation and flow cytometric quality control of clinical-scale TCRαβ/CD19-depleted grafts.
abstract: The depletion of TCRαβ+T cells and CD19+B cells is a graft purification method for haploidentical stem cell transplantation (HSCT) retaining stem cells, NK cells and TCRγδ+T cells. To avoid treatment-related occurrence of severe GvHD a precise quantification of residual TCRαβ+T cells in the graft is of essential importance.
  
  
  
  Nine stem cell grafts were purified immunomagnetically on a CliniMACS device and flow cytometric quality control (QC) was performed before and after TCRαβ/CD19-depletion.
  
  
  
  As a challenge a new 10-color QC-panel was established, which enables accurate quantification of the graft composition. The binding sites of residual TCRαβ+T and CD19+B cells were at least partly occupied by depletion antibodies impeding flow cytometric analysis. Based on respective controls and an assumed variation coefficient of 18%, the detection limit of residual TCRαβ T cells was 1 cell/µl. and 0.002% of CD45+cells. Log-depletion of TCRαβ and CD19 cells was -3.9 and -3.3, respectively. The recovery was 82.1%, 67.1% and 72.7% for stem cells, NK cells and for TCRγδ+T cells.
  
  
  
  In clinical use this method may help to improve transplantation outcome, due to the correct application of the desired stem cell and the limited T cell dose. The panel is designed for the QC following TCRαβ/CD19-depletion but is adaptable to other depletion strategies as well. © 2015 International Clinical Cytometry Society.
authors.affiliation: Clinic for Pediatric and Adolescent Medicine, University Hospital, Frankfurt, Germany.
authors.forename: Melanie
authors.initials: M
authors.lastname: Bremm
authors.affiliation: Clinic for Pediatric and Adolescent Medicine, University Hospital, Frankfurt, Germany.
authors.forename: Claudia
authors.initials: C
authors.lastname: Cappel
authors.affiliation: Clinic for Pediatric and Adolescent Medicine, University Hospital, Frankfurt, Germany.
authors.forename: Stephanie
authors.initials: S
authors.lastname: Erben
authors.affiliation: Clinic for Pediatric and Adolescent Medicine, University Hospital, Frankfurt, Germany.
authors.forename: Andrea
authors.initials: A
authors.lastname: Jarisch
authors.affiliation: Department of Hematology/Oncology, Children's University Hospital, Tübingen, Germany.
authors.forename: Michael
authors.initials: M
authors.lastname: Schumm
authors.affiliation: Miltenyi Biotec GmbH, Bergisch Gladbach, Germany.
authors.forename: Andreas
authors.initials: A
authors.lastname: Arendt
authors.affiliation: Institute for Transfusion Medicine and Immunohematology, Goethe-University Frankfurt/Main, Division for Cell Processing, German Red Cross Blood Donor Service Baden-Württemberg-Hessen, Frankfurt/Main, Germany.
authors.forename: Halvard
authors.initials: H
authors.lastname: Bonig
authors.affiliation: Clinic for Pediatric and Adolescent Medicine, University Hospital, Frankfurt, Germany.
authors.forename: Thomas
authors.initials: T
authors.lastname: Klingebiel
authors.affiliation: Institute of Cellular Therapeutics, GMP Development Unit, IFB-TX; Hannover Medical School, Hannover, Germany.
authors.forename: Ulrike
authors.initials: U
authors.lastname: Koehl
authors.affiliation: Clinic for Pediatric and Adolescent Medicine, University Hospital, Frankfurt, Germany.
authors.forename: Peter
authors.initials: P
authors.lastname: Bader
authors.affiliation: Clinic for Pediatric and Adolescent Medicine, University Hospital, Frankfurt, Germany.
authors.forename: Sabine
authors.initials: S
authors.lastname: Huenecke
mesh_terms: D018941:Antigens, CD19 / Q000276:immunology*; D001402:B-Lymphocytes / Q000166:cytology* / Q000276:immunology; D017252:CD3 Complex / Q000276:immunology; D002452:Cell Count / Q000379:methods; D005434:Flow Cytometry; D006086:Graft vs Host Disease / Q000276:immunology*; D018380:Hematopoietic Stem Cell Transplantation / Q000379:methods; D006801:Humans; D007694:Killer Cells, Natural / Q000166:cytology* / Q000276:immunology; D011786:Quality Control; D016693:Receptors, Antigen, T-Cell, alpha-beta / Q000276:immunology*; D013601:T-Lymphocytes / Q000166:cytology* / Q000276:immunology
chemical_list: D018941:Antigens, CD19; C000604811:CD19 molecule, human; D017252:CD3 Complex; D016693:Receptors, Antigen, T-Cell, alpha-beta
keywords: TCRαβ/CD19-depletion; children; flow cytometry; stem cell transplantation