GMP-production of purified human B lymphocytes for the adoptive transfer in patients after allogeneic hematopoietic stem cell transplantation

Beitrag in einer Fachzeitschrift


Details zur Publikation

Autor(en): Tittlbach H, Schneider A, Strobel J, Zimmermann R, Maas S, Gebhardt B, Rauser G, Mach M, Mackensen A, Winkler T, Winkler J
Zeitschrift: Journal of Translational Medicine
Jahr der Veröffentlichung: 2017
Band: 15
Heftnummer: 1
Seitenbereich: 228
ISSN: 1479-5876


Abstract


We have recently shown that memory B cells from murine CMV immune donor animals adoptively transferred into immunodeficient mice were highly effective in protecting from a viral infection indicating a therapeutic potential of virus specific memory B cells. These preclinical data provided evidence that a cell-based strategy supporting the humoral immune response might be effective in a clinical setting of immunodeficiency after allogeneic hematopoietic stem cell transplantation. As adoptive transfer of B cells has not been used before in a clinical setting it was necessary to establish a technology for the generation of good manufacturing practice (GMP)-grade B cell products.Starting from the leukapheresis product of healthy blood donors, B cells were purified by two different separation strategies using GMP-grade microbeads and the CliniMACS system. A one-step protocol was used for positive enrichment of B lymphocytes with anti-CD19 microbeads. In a two-step enrichment protocol, first T lymphocytes were depleted by anti-CD3 microbeads and the remaining fraction was positively selected by anti-CD19 microbeads.The purity and recovery after enrichment of B lymphocytes from the leukapheresis material in both separations strategies was not statistically different. However, contamination of the B-cell product with T cells was significantly lower after the two-step protocol (0.16%, range 0.01-0.43% after two-step separation and 0.55%, range 0.28-0.85% after one-step separation, p < 0.05). Therefore, a combined CD3 depletion and CD19 enrichment was used for the production of GMP-conform B-cell products from the leukapheresis material of 17 healthy stem cell donors. The absolute B-cell numbers obtained in the final product was 4.70 ± 3.64 × 108 with a purity of 95.98 ± 3.31% B lymphocytes and a recovery of 18.9 ± 10.6%. Importantly, the contamination with CD3+ T cells was extremely low in the final B- cell products (0.10 ± 0.20%). Purified B cells exhibited normal antibody production after in vitro stimulation and showed excellent viability after cryopreservation.A GMP-grade B-cell product can be obtained with high purity and very low T-cell contamination using the two-step enrichment protocol based on CliniMACS® technology.



FAU-Autoren / FAU-Herausgeber

Mach, Michael apl. Prof. Dr.
Medizinische Fakultät
Mackensen, Andreas Prof. Dr.
Lehrstuhl für Innere Medizin V
Schneider, Andrea
Lehrstuhl für Genetik
Strobel, Joachim
Institut für Experimentelle und Klinische Pharmakologie und Toxikologie
Winkler, Jürgen Prof. Dr.
Molekular-Neurologische Abteilung in der Neurologischen Klinik
Winkler, Thomas Prof. Dr.
Professur für Genetik
Zimmermann, Robert Prof. Dr.
Medizinische Fakultät


Autor(en) der externen Einrichtung(en)
Miltenyi Biotec GmbH


Zitierweisen

APA:
Tittlbach, H., Schneider, A., Strobel, J., Zimmermann, R., Maas, S., Gebhardt, B.,... Winkler, J. (2017). GMP-production of purified human B lymphocytes for the adoptive transfer in patients after allogeneic hematopoietic stem cell transplantation. Journal of Translational Medicine, 15(1), 228. https://dx.doi.org/10.1186/s12967-017-1330-5

MLA:
Tittlbach, Hannes, et al. "GMP-production of purified human B lymphocytes for the adoptive transfer in patients after allogeneic hematopoietic stem cell transplantation." Journal of Translational Medicine 15.1 (2017): 228.

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Zuletzt aktualisiert 2018-08-10 um 08:28