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72. Jahrestagung der Deutschen Gesellschaft für Neurochirurgie (DGNC)
Joint Meeting mit der Polnischen Gesellschaft für Neurochirurgie

Deutsche Gesellschaft für Neurochirurgie (DGNC) e. V.

06.06. - 09.06.2021

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Allogeneic NKG2C-positive NK cells for immunotherapy of glioblastoma multiforme

Allogene NKG2C-positive NK-Zellen zur Immuntherapie des Glioblastoma multiforme

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  • presenting/speaker Shafiq Murad - University Hospital Carl Gustav Carus, Department of Neurosurgery, Section Experimental Neurosurgery and Tumor Immunology, Dresden, Deutschland
  • Susanne Michen - University Hospital Carl Gustav Carus, Department of Neurosurgery, Section Experimental Neurosurgery and Tumor Immunology, Dresden, Deutschland
  • Alexander Becker - University Hospital Carl Gustav Carus, Department of Neurosurgery, Section Experimental Neurosurgery and Tumor Immunology, Dresden, Deutschland
  • André Sagerer - University Hospital Carl Gustav Carus, Department of Neurosurgery, Section Experimental Neurosurgery and Tumor Immunology, Dresden, Deutschland
  • Monika Füssel - DKMS Life Science Lab GmbH, Dresden, Deutschland
  • Gabriele Schackert - University Hospital Carl Gustav Carus, Department of Neurosurgery, Section Experimental Neurosurgery and Tumor Immunology, Dresden, Deutschland; German Cancer Consortium (DKTK), partner site Dresden, Dresden, Deutschland; National Center for Tumor Diseases (NCT), DKFZ partner site Dresden, Dresden, Deutschland
  • Frank Momburg - German Cancer Research Center (DKFZ), Clinical Cooperation Unit Applied Tumor Immunity, Antigen Presentation & T/NK Cell Activation Group, Heidelberg, Deutschland
  • Achim Temme - University Hospital Carl Gustav Carus, Department of Neurosurgery, Section Experimental Neurosurgery and Tumor Immunology, Dresden, Deutschland; German Cancer Consortium (DKTK), partner site Dresden, Dresden, Deutschland; National Center for Tumor Diseases (NCT), DKFZ partner site Dresden, Dresden, Deutschland

Deutsche Gesellschaft für Neurochirurgie. 72. Jahrestagung der Deutschen Gesellschaft für Neurochirurgie (DGNC), Joint Meeting mit der Polnischen Gesellschaft für Neurochirurgie. sine loco [digital], 06.-09.06.2021. Düsseldorf: German Medical Science GMS Publishing House; 2021. DocV291

doi: 10.3205/21dgnc276, urn:nbn:de:0183-21dgnc2764

Published: June 4, 2021

© 2021 Murad et al.
This is an Open Access article distributed under the terms of the Creative Commons Attribution 4.0 License. See license information at http://creativecommons.org/licenses/by/4.0/.

Outline

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Objective: NKG2C-positive natural killer (NK) cells represents a small NK cell subset, which is found in higher frequency in human cytomegalovirus (HCMV) seropositive patients. NKG2C+ NK cells can recognize target cells displaying HLA-E loaded with intracellularly processed ER signal peptides (sp) derived from classical HLA-A,B,C alleles and from HLA-G. Of note, HLA-E and HLA-G is frequently expressed in glioblastoma multiforme (GBM). Consequently, the NKG2C+ NK subset might have an increased intrinsic capacity to recognize and kill GBM cells. This study investigated, whether allogeneic NKG2C+ NK cells have an anti-tumor effect against HLA-E+/G+ GBM cells.

Methods: GBM patients and NK donors were HLA genotyped for HLA:KIR match/mismatch analysis. HCMV IgG and IgM titers from NK cell donors were analyzed using ELISA. HLA-E+/G+ GBM cells were prepared by enzymatic/mechanic tumor dissociation and were cultivated in standard DMEM medium. Untouched NK cells were generated using negative NK cell isolation technique. The NKG2C+ NK cell fraction was expanded for 14 days using NK feeder cells, designated, PC3-PSCA-IL-2-mIL-15d-HLA-EspG, expressing an activating B2M-HLA-EspG trimeric fusion protein, IL-2 and membrane-bound IL-15. Enrichment of expanded NKG2C+/A- NK cells was accomplished by MACS-assisted depletion of NKG2A+ NK cells. For control, NK cells from same donors were expanded using a feeder cell line only expressing IL-2 and mIL-15d known to augment outgrowth of NKG2A+/C- NK cells. Freshly prepared and expanded NK cells were characterized using antibodies for CD56, NKG2A, NKG2C and KIRs. Cytotoxicity of NK cells towards GBM cells was investigated by chromium release assay.

Results: PC3-PSCA-IL-2-mIL-15d-HLA-EspG feeder cells induced a 32-fold increase in NKG2C+ NK cell numbers. Likewise, the mean frequency of NKG2C+ NK cells in the NK cell population increased from in the mean 9% to 33%. MACS-assisted depletion of NKG2A+ NK cells resulted in NKG2C+ NK cell purity >90%. In cytotoxicity assays, only NKG2C+ NK cells showed lytic activity towards HLA-E/G-positive GBM cells. Of note, NKG2C+ NK cells were reactive against C2/Bw4-mismatched but also towards KIR:HLA-matched allogeneic GBM cells.

Conclusion: Our results demonstrate the feasibility of allogeneic NKG2C+ NK cells for immunotherapy of GBM, which operates independently from KIR/HLA-setting. Further preclinical studies are required to confirm the potential of NKG2C+ NK cells for immunotherapy of glioblastoma.