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Xhaxho, S.* ; Chen-Wichmann, L.* ; Kreissig, S.* ; Windisch, R.* ; Gottschlich, A.* ; Nandi, S.* ; Schabernack, S.* ; Kohler, I.* ; Kellner, C.* ; Kobold, S. ; Humpe, A.* ; Wichmann, C.*

Efficient chimeric antigen receptor T-cell generation starting with leukoreduction system chambers of thrombocyte apheresis sets.

Transfus. Med. Hemother. 51, 111-118 (2023)
Verlagsversion DOI PMC
Open Access Gold
Creative Commons Lizenzvertrag
Introduction: Primary human blood cells represent an essential model system to study physiology and disease. However, human blood is a limited resource. During healthy donor plateletpheresis, the leukoreduction system chamber (LRSC) reduces the leukocyte amount within the subsequent platelet concentrate through saturated, fluidized, particle bed filtration technology. Normally, the LRSC is discarded after apheresis is completed. Compared to peripheral blood, LRSC yields 10-fold mononuclear cell concentration. Methods: To explore if those retained leukocytes are attractive for research purposes, we isolated CD3+ T cells from the usually discarded LRSCs via density gradient centrifugation in order to manufacture CD19-targeted chimeric antigen receptor (CAR) T cells. Results: Immunophenotypic characterization revealed viable and normal CD4+ and CD8+ T-cell populations within LRSC, with low CD19+ B cell counts. Magnetic-activated cell sorting (MACS) purified CD3+ T cells were transduced with CD19 CAR-encoding lentiviral self-inactivating vectors using concentrated viral supernatants. Robust CD19 CAR cell surface expression on transduced T cells was confirmed by flow cytometry. CD19 CAR T cells were further enriched through anti-CAR MACS, yielding 80% CAR+ T-cell populations. In vitro CAR T cell expansion to clinically relevant numbers was achieved. To prove functionality, CAR T cells were co-incubated with the human CD19+ B cell precursor leukemia cell line Nalm6. Compared to unmodified T cells, CD19 CAR T cells effectively eradicated Nalm6 cells. Conclusion: Taken together, we can show that lymphocytes isolated from LRSCs of plateletpheresis sets can be efficiently used for the generation of functional CAR T cells for experimental purposes.
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Publikationstyp Artikel: Journalartikel
Dokumenttyp Wissenschaftlicher Artikel
Schlagwörter Cell Therapy ; Chimeric Antigen Receptor T Cell ; Leukoreduction System Chamber ; T Cell; Blood Mononuclear-cells; Quality
ISSN (print) / ISBN 1660-3796
e-ISSN 1424-5493
Zeitschrift Transfusion Medicine and Hemotherapy
Quellenangaben Band: 51, Heft: 2, Seiten: 111-118 Artikelnummer: , Supplement: ,
Verlag Karger
Verlagsort Allschwilerstrasse 10, Ch-4009 Basel, Switzerland
Begutachtungsstatus Peer reviewed
Institut(e) Unit for Clinical Pharmacology (KKG-EKLiP)
Förderungen Stiftung Transfusionsmedizin und Immunhaematologie
"i-Target: immunotargeting of cancer" ( - Elite Network of Bavaria)
Melanoma Research Alliance
Else Kroner-Fresenius-Stiftung, German Cancer Aid
Wilhelm-Sander-Stiftung
Ernst Jung Stiftung
Institutional Strategy LMU
Go-Bio-Initiative
Else Kroener-Fresenius-Stiftung
Bundesministerium fur Bildung und Forschung
European Research Council
Deutsche Forschungsgemeinschaft (DFG)
SFB-TRR
Fritz -Bender Foundation
Deutsche Jose Carreras Leukaemie Stiftung
Hector Foundation
m4-Award of the Bavarian Ministry for Economic Affairs