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Original Research ARTICLE Provisionally accepted The full-text will be published soon. Notify me

Front. Immunol. | doi: 10.3389/fimmu.2019.01218

Improving clinical manufacturing of IL-15 activated cytokine-induced killer (CIK) cells

 Melanie Bremm1, 2*, Claudia Cappel1, 2, Verena Katzki1, 2, Stephanie Erben1, 2, Sibille Betz1, 2, Andrea Quaiser3,  Halvard Bonig4, Michael Schmidt4,  Lisa-Marie Pfeffermann1, 2,  Thomas E. Klingebiel1, 2,  Peter Bader1, 2,  Sabine Huenecke1, 2 and  Eva Rettinger1, 2
  • 1University Hospital Frankfurt, Germany
  • 2Klinik für Kinder und Jugendmedizin, Universitätsklinikum Frankfurt, Germany
  • 3Fraunhofer-Institut für Zelltherapie und Immunologie (IZI), Germany
  • 4Institute for Transfusion Medicine and Immunohematology, German Red Cross Blood Donor Services, Germany

Cytokine-induced killer (CIK) cells are an immunotherapeutic approach to combat relapse following allogeneic hematopoietic stem cell transplantation (HSCT) in acute leukemia or myelodysplastic syndrome (MDS) patients. Prompt and sequential administration of escalating cell doses improves the efficacy of CIK cell therapy without exacerbating graft versus host disease (GVHD).
This study addresses manufacturing-related issues and aimed to develop a time-, personal- and cost-saving good manufacturing process (GMP)-compliant protocol for the generation of ready-for-use therapeutic CIK cell doses starting from one unstimulated donor-derived peripheral blood (PB) or leukocytapheresis (LP) products. Culture medium with or without the addition of either AB serum, fresh frozen plasma (FFP) or platelet lysate (PL) was used for culture. Fresh and cryopreserved CIK cells were compared regarding expansion rate, viability, phenotype, and ability to inhibit leukemia growth.
Cell numbers increased by a median factor of 10-fold in the presence of FFP, PL or AB serum, whereas cultivation in FFP/PL-free or AB serum-free medium failed to promote adequate CIK cell proliferation (p<0.01) needed to provide clinical doses of 1x106 T cells/kG, 5x106 T cells/kG, 1x107 T cells/kG, and 1x108 T cells/kG recipient body weight. CIK cells consisting of T cells, T- natural killer (T-NK) cells and a minor fraction of NK cells were not significantly modified by different medium supplements. Moreover, neither cytotoxic potential against leukemic THP-1 cells nor cell activation shown by CD25 expression were significantly influenced. Moreover, overnight and long-term cryopreservation had no significant effect on the composition of CIK cells, their phenotype or cytotoxic potential. A viability of almost 93% (range: 89 - 96) and 89.3% (range: 84 – 94) was obtained after freeze-thawing procedure and long-term storage, respectively, whereas viability was 96% (range: 90 – 97) in fresh CIK cells.
Altogether, GMP-complaint CIK cell generation from an unstimulated donor-derived PB or LP products was feasible. Introducing FFP, which is easily accessible, into CIK cell cultures was time- and cost-saving without loss of viability and potency in a 10-12 day batch culture. The feasibility of cryopreservation enabled storage and delivery of sequential highly effective ready-for-use CIK cell doses and therefore reduced the number of manufacturing cycles.

Keywords: CIK cells, Immunotherapy, allogeneic stem cell transplantation, Cryopreservation, AB-serum, Fresh frozen plasma, Platelet lysate

Received: 15 Feb 2019; Accepted: 13 May 2019.

Edited by:

Aurore Saudemont, GlaxoSmithKline (United Kingdom), United Kingdom

Reviewed by:

Dario Sangiolo, University of Turin, Italy
Ingo Schmidt-Wolf, Universitätsklinikum Bonn, Germany  

Copyright: © 2019 Bremm, Cappel, Katzki, Erben, Betz, Quaiser, Bonig, Schmidt, Pfeffermann, Klingebiel, Bader, Huenecke and Rettinger. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

* Correspondence: Dr. Melanie Bremm, University Hospital Frankfurt, Frankfurt, Hesse, Germany,