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. 2022 Sep 27;10(9):e005187. doi: 10.1136/jitc-2022-005187

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© Author(s) (or their employer(s)) 2022. Re-use permitted under CC BY-NC. No commercial re-use. See rights and permissions. Published by BMJ.

This is an open access article distributed in accordance with the Creative Commons Attribution Non Commercial (CC BY-NC 4.0) license, which permits others to distribute, remix, adapt, build upon this work non-commercially, and license their derivative works on different terms, provided the original work is properly cited, appropriate credit is given, any changes made indicated, and the use is non-commercial. See http://creativecommons.org/licenses/by-nc/4.0/.

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High-level GD2 expression in glioblastoma (GBM) tumor tissues and glioma neural stem (GNS) cell lines, but not in normal brain. (A) Sections of surgical specimens from GBM (n=16) or surrounding non-involved brain (n=4) were stained by immunofluorescence using an anti-GD2 primary antibody (clone 14g2a) and IgG2a isotype control antibody (top row insets). Representative staining for regions of high (top) and low (middle) GD2 expression, and matched adjacent normal brain tissue (bottom). (B) Summary of GD2 staining intensity measured by ImageJ. Dotted lines at y-axis mark average staining intensity for (1) adjacent normal brain tissue (n=4) removed by neurosurgeon to access the tumor, and (2) the isotype control. (C) Summary of GD2 expression on GNS cell lines, which were generated from patients with GBM and DIPG’s tumors and maintained in culture for <25 passages. (D) Representative histograms showing the three distinct GD2 expression patterns observed. Cells were analyzed by flow cytometry for GD2 expression using anti-GD2 primary mAb (clone 14g2a; black histograms) or isotype-matched control antibodies (red histograms). (E) The CCB-G5C GNS cell line was implanted in the brains of NOD-SCID-gamma-null (NSG) mice via stereotactic intracranial injection. Representative image of H&E staining (left) and GD2 immunofluorescence (right) of coronal section of mouse brain at time of humane killing because of neurological signs n=10, for full analysis of groups see figures 4–5. Asterisk marks side of tumor inoculation. CCB, Centre for Cancer Biology; DIPG, diffuse intrinsic pontine glioma.