Media type: E-Article Title: Generation and flow cytometric quality control of clinical‐scale TCRαβ/CD19‐depleted grafts Contributor: Bremm, Melanie; Cappel, Claudia; Erben, Stephanie; Jarisch, Andrea; Schumm, Michael; Arendt, Andreas; Bonig, Halvard; Klingebiel, Thomas; Koehl, Ulrike; Bader, Peter; Huenecke, Sabine Published: Wiley, 2017 Published in: Cytometry Part B: Clinical Cytometry, 92 (2017) 2, Seite 126-135 Language: English DOI: 10.1002/cyto.b.21328 ISSN: 1552-4957; 1552-4949 Origination: Footnote: Description: BackgroundThe 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.MethodsNine stem cell grafts were purified immunomagnetically on a CliniMACS device and flow cytometric quality control (QC) was performed before and after TCRαβ/CD19‐depletion.ResultsAs 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.ConclusionsIn 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 Access State: Open Access