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Interleukin-2-stimulated natural killer cells are less susceptible to mycophenolate mofetil than non-activated NK cells: possible consequences for immunotherapy

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Abstract

In a clinical phase I/II trial, pediatric patients with high-risk malignancies were treated with ex vivo IL-2-stimulated donor natural killer (NK) cells after transplantation with haploidentical stem cells. To evaluate the potential negative effects of the immunosuppressive drug mycophenolate mofetil (MMF) used for immunotherapy, the functionality and signaling of ex vivo NK cells was investigated. Our results show that during NK cell expansion, long-term (9 days) incubation with mycophenolic acid (MPA), the active metabolite of MMF, in therapeutically relevant concentrations led to the severe inhibition of NK cell proliferation. This correlated with a significantly reduced cytokine/chemokine secretion and the inhibited acquisition of surface receptors regarding cytotoxicity (e.g., NKp30, NKp44, NKp46, NKG2D), adhesion/migration (e.g., ICAM-1/CD54, LFA-1/CD11a, CD62L, CXCR3) and activation (e.g., CD25). Moreover, MPA prevented phosphorylation of the central signaling molecules STAT-3/-4/-5, AKT and ERK1/2. In contrast, short-term (24 h) MPA incubation of IL-2-stimulated NK cells had no or only marginal effects on the activated NK cell phenotype, including receptor expression, cytokine/chemokine secretion and intracellular signaling. Further, short-term MPA incubation only moderately affected the highly cytotoxic activity of previously IL-2-stimulated NK cells. In conclusion, while long-term MPA incubation significantly compromised ex vivo NK cell functionality, previously IL-2-activated NK cells seemed to be rather resistant to short-term MPA treatment. This finding supports the use of IL-2-activated NK cells as immunotherapy, especially for patients treated with MMF after haploidentical stem cell transplantation.

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Abbreviations

BW:

Body weight

GCP:

Good clinical practice

GMP:

Good manufacturing practice

GvL/T:

Graft-versus-leukemia/tumor

GvHD:

Graft-versus-host-disease

haplo-SCT:

Haploidentical stem cell transplantation

IMPDH:

Inosine monophosphate dehydrogenase

MFI:

Mean fluorescence intensity

MMF:

Mycophenolate mofetil

MPA:

Mycophenolic acid

NB:

Neuroblastoma

NCR:

Natural cytotoxicity receptors

NK-DLI:

Natural killer cell donor lymphocyte infusion

NKG2D:

NK group 2D

PB:

Peripheral blood

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Acknowledgments

This project was supported by the “Deutsche Forschungsgemeinschaft (DFG)“(GRK-1172), “Hilfe für krebskranke Kinder Frankfurt e.V.” and the “LOEWE Center for Cell and Gene Therapy Frankfurt” funded by Hessisches Ministerium für Wissenschaft und Kunst, III L 4- 518/17.004 (2010). The authors would like to thank all patients, nurses and physicians and the members of the Laboratory of Stem Cell Transplantation and Immunotherapy for their technical support and Dr. D. Hintereder of the Central Laboratory for the quantification of total MPA in patients’ PB plasma.

Conflict of interest

The authors declare that they have no conflicts of interest.

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Authors and Affiliations

  1. Clinic for Pediatric and Adolescent Medicine, University Hospital Frankfurt, Goethe University, Frankfurt/Main, Germany

    Claudia Brehm, Sabine Huenecke, Andrea Quaiser, Sibille Betz, Olga Zimmermann, Jan Soerensen, Thomas Klingebiel, Dirk Schwabe & Peter Bader

  2. Institute of Cellular Therapeutics GMP Development Unit, IFB-Tx and Cellular Therapy Center, Medical School Hannover, Hannover, Germany

    Ruth Esser, Stephan Kloess & Ulrike Koehl

  3. Department of Hematology, University Hospital Basel, Basel, Switzerland

    Jakob R. Passweg

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  1. Claudia Brehm
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Correspondence to Claudia Brehm.

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Brehm, C., Huenecke, S., Esser, R. et al. Interleukin-2-stimulated natural killer cells are less susceptible to mycophenolate mofetil than non-activated NK cells: possible consequences for immunotherapy. Cancer Immunol Immunother 63, 821–833 (2014). https://doi.org/10.1007/s00262-014-1556-5

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