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01.12.2018 | Methodology | Ausgabe 1/2018 Open Access

Journal of Translational Medicine 1/2018

Clinical-grade generation of peptide-stimulated CMV/EBV-specific T cells from G-CSF mobilized stem cell grafts

Journal of Translational Medicine > Ausgabe 1/2018
Regina Gary, Michael Aigner, Stephanie Moi, Stefanie Schaffer, Anja Gottmann, Stefanie Maas, Robert Zimmermann, Jürgen Zingsem, Julian Strobel, Andreas Mackensen, Josef Mautner, Andreas Moosmann, Armin Gerbitz
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Electronic supplementary material

The online version of this article (https://​doi.​org/​10.​1186/​s12967-018-1498-3) contains supplementary material, which is available to authorized users.
Regina Gary and Michael Aigner contributed equally to this work
Andreas Moosmann and Armin Gerbitz contributed equally to this work



A major complication after allogeneic hematopoietic stem cell transplantation (aSCT) is the reactivation of herpesviruses such as cytomegalovirus (CMV) and Epstein–Barr virus (EBV). Both viruses cause significant mortality and compromise quality of life after aSCT. Preventive transfer of virus-specific T cells can suppress reactivation by re-establishing functional antiviral immune responses in immunocompromised hosts.


We have developed a good manufacturing practice protocol to generate CMV/EBV-peptide-stimulated T cells from leukapheresis products of G-CSF mobilized and non-mobilized donors. Our procedure selectively expands virus-specific CD8+ und CD4+ T cells over 9 days using a generic pool of 34 CMV and EBV peptides that represent well-defined dominant T-cell epitopes with various HLA restrictions. For HLA class I, this set of peptides covers at least 80% of the European population.


CMV/EBV-specific T cells were successfully expanded from leukapheresis material of both G-CSF mobilized and non-mobilized donors. The protocol allows administration shortly after stem cell transplantation (d30+), storage over liquid nitrogen for iterated applications, and protection of the stem cell donor by avoiding a second leukapheresis.


Our protocol allows for rapid and cost-efficient production of T cells for early transfusion after aSCT as a preventive approach. It is currently evaluated in a phase I/IIa clinical trial.
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