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Technical choices significantly alter the adaptive immune response against immunocompetent murine gliomas in a model-dependent manner

  • Laboratory Investigation
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Journal of Neuro-Oncology Aims and scope Submit manuscript

Abstract

Purpose

Due to the recent rise in immunotherapy research to treat glioblastoma (GBM), immunocompetent mouse models have become increasingly crucial. However, the character and kinetics of the immune response against the most prevalent immunocompetent GBM models, GL261 and CT2A, have not been well studied, nor has the impact of commonly-used marker proteins and foreign antigens.

Methods

In this study, we compared the immune response in these models using flow cytometry and immunohistochemistry as well as investigated several factors that influence the immune response, including kinetics, tumor size, and expression of commonly-used marker proteins and foreign antigens. We hypothesize that these factors influence the immune response enough to warrant consideration when studying new immunotherapeutic approaches for GBM.

Results

CT2A-Luc, but not GL261-Luc2, drastically increased the number of T cells in the brain compared with wild-type controls, and significantly altered CT2A’s responsiveness to anti-PD-1 antibody therapy. Additionally, a larger cell inoculum size in the GL261 model increased the T cell response’s magnitude at day 28 post-injection. CT2A and GL261 models both stimulate a peak T cell immune response at day 21 post-injection.

Conclusions

Our results suggest that the impact of foreign proteins like luciferase on the intracranial immune response is dependent upon the model, with CT2A being more sensitive to added markers. In particular, luciferase expression in CT2A could lead to meaningful misinterpretations of results from immune checkpoint inhibitor (ICI) studies.

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Data availability

Datasets generated during the current study are available from the corresponding author on reasonable request.

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Acknowledgements

Thanks to the UVA Molecular Imaging Core, the Flow Cytometry Core, Biorepository and Tissue Research Core, and the Research Histology Core for their assistance.

Funding

Thanks to the Schiff Foundation, National Institute of Health Grants F99CA234954, R01CA180699, and R21NS108057 for funding.

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Authors and Affiliations

  1. Department of Pathology, University of Virginia, Charlottesville, VA, USA

    Breanna Noffsinger, Alexandra Witter, Laryssa Manigat & Tim Bullock

  2. Department of Biomedical Engineering, University of Virginia, Charlottesville, VA, USA

    Natasha Sheybani & Richard Price

  3. Department of Neurology, University of Virginia, Charlottesville, VA, USA

    Aizhen Xiao, Qing Zhong & Benjamin Purow

  4. Department of Neuroscience, University of Virginia, Charlottesville, VA, USA

    Suchet Taori & Tajie Harris

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  1. Breanna Noffsinger
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Contributions

All authors contributed to the study conception and design. Material preparation, data collection and analysis were performed by BN, AW, NS, AX, LM, QZ, and ST. The first draft of the manuscript was written by BN and all authors commented on previous versions of the manuscript. All authors read and approved the final manuscript.

Corresponding author

Correspondence to Benjamin Purow.

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Mouse protocols were approved by the IACUC committee at the University of Virginia (UVA).

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Noffsinger, B., Witter, A., Sheybani, N. et al. Technical choices significantly alter the adaptive immune response against immunocompetent murine gliomas in a model-dependent manner. J Neurooncol 154, 145–157 (2021). https://doi.org/10.1007/s11060-021-03822-7

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