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Glioblastoma cells potentiate the induction of the Th1-like profile in phosphoantigen-stimulated γδ T lymphocytes

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Abstract

Purpose

γδ T lymphocytes are non-conventional T cells that participate in protective immunity and tumor surveillance. In healthy humans, the main subset of circulating γδ T cells express the TCRVγ9Vδ2. This subset responds to non-peptide prenyl-pyrophosphate antigens such as (E)-4-hydroxy-3-methyl-but-enyl pyrophosphate (HMBPP). This unique feature of Vγ9Vδ2 T cells makes them a candidate for anti-tumor immunotherapy. In this study, we investigated the response of HMBPP-activated Vγ9Vδ2 T lymphocytes to glioblastoma multiforme (GBM) cells.

Methods

Human purified γδ T cells were stimulated with HMBPP (1 µM) and incubated with GBM cells (U251, U373 and primary GBM cultures) or their conditioned medium. After overnight incubation, expression of CD69 and perforin was evaluated by flow cytometry and cytokines production by ELISA. As well, we performed a meta-analysis of transcriptomic data obtained from The Cancer Genome Atlas.

Results

HMBPP-stimulated γδ T cells cultured with GBM or its conditioned medium increased CD69, intracellular perforin, IFN-γ, and TNF-α production. A meta-analysis of transcriptomic data showed that GBM patients display better overall survival when mRNA TRGV9, the Vγ9 chain-encoding gene, was expressed in high levels. Moreover, its expression was higher in low-grade GBM compared to GBM. Interestingly, there was an association between γδ T cell infiltrates and TNF-α expression in the tumor microenvironment.

Conclusion

GBM cells enhanced Th1-like profile differentiation in phosphoantigen-stimulated γδ T cells. Our results reinforce data that have demonstrated the implication of Vγ9Vδ2 T cells in the control of GBM, and this knowledge is fundamental to the development of immunotherapeutic protocols to treat GBM based on γδ T cells.

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Data availability

The datasets generated for this study are available on request to the corresponding author. The datasets employed for the transcriptomic meta-analysis in GBM samples were obtained from The Cancer Genome Atlas (TCGA): https://portal.gdc.cancer.gov.

Abbreviations

GBM:

Glioblastoma multiforme

LGG:

Low grade glioblastoma

HMBPP:

(E)-4-hydroxy-3-methyl-but-2-enyl pyrophosphate

IFN-γ:

Interferon gamma

TNF-α:

Tumor necrosis factor alpha

HGEC:

Human glomerular endothelial cells

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Acknowledgements

The authors thank Dr. Marianela Candolfi for providing the U251 and U373 cell lines. This work was supported by Grant from Agencia Nacional de Promoción Científica y Tecnológica (Grant No. PICT2016/700), Consejo Nacional de Investigaciones Científicas y Técnicas and Fundación JA Roemmers. The authors thank Instituto Nacional del Cáncer for the financial support (fellowship to David A. Rosso).

Funding

This work was supported by grant from Agencia Nacional de Promoción Científica y Tecnológica (PICT2016/700) and Consejo Nacional de Investigaciones Científicas y Técnicas and Fundación JA Roemmers.

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Author notes

  1. GabrielaV. Salamone and Carolina C. Jancic have contributed equally to this work.

Authors and Affiliations

  1. Instituto de Medicina Experimental – CONICET – Academia Nacional de Medicina, Buenos Aires, Argentina

    David A. Rosso, Micaela Rosato, Carolina M. Shiromizu, Irene A. Keitelman, Juan V. Coronel, Gabriela V. Salamone & Carolina C. Jancic

  2. División Neurocirugía, Instituto de Investigaciones Médicas A Lanari, Universidad de Buenos Aires, Buenos Aires, Argentina

    Juan Iturrizaga & Alejandra T. Rabadan

  3. Instituto de Investigaciones Biomédicas (INBIOMED) – Universidad de Buenos Aires – CONICET, Facultad de Medicina, Universidad de Buenos Aires, Buenos Aires, Argentina

    Nazareno González

  4. Laboratorio de Fisiopatogenia, Departamento de Fisiología, Facultad de Medicina, Universidad de Buenos Aires, Buenos Aires, Argentina

    Fernando D. Gómez & María M. Amaral

  5. Departamento de Microbiología, Parasitología e Inmunología, Facultad de Medicina, Universidad de Buenos Aires, Buenos Aires, Argentina

    Gabriela V. Salamone & Carolina C. Jancic

Authors
  1. David A. Rosso
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  2. Micaela Rosato
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Contributions

Material preparation, experiments, and analysis of the data: DAR and MR. GBM sample obtention: JI. Bioinformatic analysis: NG. Contributions to the manuscript writing and revision: DAR, MR, CMS, IAK, and JVC. HGEC culture: FDG and MMA. Critical revision of the manuscript: ATR. Data interpretation and results discussion: GVS and CCJ. Experimental design and manuscript writing: CCJ. The authors read and approved the manuscript.

Corresponding author

Correspondence to Carolina C. Jancic.

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This study was reviewed and approved by the ethical committee of the Institutos de la Academia Nacional de Medicina, Instituto de Investigaciones Médicas A. Lanari, and Universidad de Buenos Aires. The procedures used in this study adhere to the tenets of the Declaration of Helsinki.

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Rosso, D.A., Rosato, M., Iturrizaga, J. et al. Glioblastoma cells potentiate the induction of the Th1-like profile in phosphoantigen-stimulated γδ T lymphocytes. J Neurooncol 153, 403–415 (2021). https://doi.org/10.1007/s11060-021-03787-7

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