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NKG2D ligands in glioma stem-like cells: expression in situ and in vitro

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Abstract

Glioblastoma multiforme (GBM) is a highly malignant brain tumor. Tumor stem cells have a major influence on tumor malignancy, and immunological escape mechanisms, involving the Natural Killer Group 2, member D (NKG2D) receptor-ligand-system, are key elements in tumor immuno-surveillance. We analyzed the expression profile and localization of NKG2D ligands (NKG2DL) and embryonic and neural stem cell markers in solid human GBM and stem-like cells isolated from glioma cell lines by qRT-PCR and immunohistochemistry, including quantitative analysis. We also evaluated the effect of Temozolomide (TMZ), the standard chemotherapeutic agent used in GBM therapy, on NKG2DL expression. NKG2DL-positive cells were mostly found scattered and isolated, were detectable in glial fibrillary acidic protein (GFAP)-positive tumor regions and partly in the penumbra of tumor vessels. NKG2DL were found in a distinct tumor stem-like cell subpopulation and were broadly costained with each other. Quantitative analysis revealed, that dependent on the individual NKG2DL investigated, cell portions costained with different stem cell markers varied between small (Musashi-1) and high (KLf-4) amounts. However, a costaining of NKG2DL with CD3γ, typically found in T cells, was also observable, whereas CD11b as a marker for tumor micoglia cells was only rarely costained with NKG2DL. Stem-like cells derived from the glioma cell lines T98G and U251MG showed a distinct expression pattern of NKG2DL and stem cell markers, which seemed to be balanced in a cell line-specific way. With differentiation, T98G displayed less NKG2DL, whereas in U251MG, only expression of most stem cell markers decreased. In addition, stimulation with TMZ led to a significant upregulation of NKG2DL in stem-like cells of both lines. As stem-like glioma cells tend to show a higher expression of NKG2DL than more differentiated tumor cells and TMZ treatment supports upregulation of NKG2DL, the NKG2D system might play an important role in tumor stem cell survival and in GBM therapy.

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Acknowledgements

We thank Fereshteh Ebrahim and Brigitte Rehmke for expert technical support.

Funding

This work was supported by a sponsorship of the University Medical Center Schleswig–Holstein UKSH (Forschungsförderung 2016), which was given to CF, by the RTG2154 [project P7 (KHK) and P8 (JHF)] and by the popgen 2.0 network [P2N; supported by a grant from the German Ministry for Education and Research (01EY1103)].

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Author notes

  1. Charlotte Flüh and Guranda Chitadze have contributed equally.

Authors and Affiliations

  1. Department of Neurosurgery, University Medical Center Schleswig-Holstein, Campus Kiel, Arnold-Heller-Straße 3, Haus 41, 24105, Kiel, Germany

    Charlotte Flüh, Vivian Adamski, Michael Synowitz & Janka Held-Feindt

  2. Institute of Immunology, University Medical Center Schleswig-Holstein, Campus Kiel, Arnold-Heller-Straße 3, Haus 17, 24105, Kiel, Germany

    Guranda Chitadze & Dieter Kabelitz

  3. Institute of Anatomy, Christian-Albrechts-University, Olshausenstraße 40, 24098, Kiel, Germany

    Kirsten Hattermann

Authors
  1. Charlotte Flüh
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  2. Guranda Chitadze
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Correspondence to Charlotte Flüh.

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Flüh, C., Chitadze, G., Adamski, V. et al. NKG2D ligands in glioma stem-like cells: expression in situ and in vitro. Histochem Cell Biol 149, 219–233 (2018). https://doi.org/10.1007/s00418-018-1633-5

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