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Therapeutic vaccination targeting CD40 and TLR3 controls melanoma growth through existing intratumoral CD8 T cells without new T cell infiltration

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Abstract

Dendritic cells are potently activated by the synergistic action of CD40 stimulation in conjunction with signaling through toll like receptors, subsequently priming T cells. Cancer vaccines targeting the activation of dendritic cells in this manner show promise in murine models and are being developed for human patients. While the efficacy of vaccines based on CD40 and toll like receptor stimulation has been established, further investigation is needed to understand the mechanism of tumor control and how vaccination alters tumor infiltrating immune cells. In this study we vaccinated mice bearing established murine melanoma tumors with agonistic anti-CD40, polyI:C, and tumor antigen. Vaccination led to increased intratumoral T cell numbers and delayed tumor growth, yet did not require trafficking of T cells from the periphery. Pre-existing intratumoral T cells exhibited an acute burst in proliferation but became less functional in response to vaccination. However, the increased intratumoral T cell numbers yielded increased numbers of effector T cells per tumor. Together, our data indicate that the existing T cell response and intratumoral dendritic cells are critical for vaccination efficacy. It also suggests that circulating T cells responding to vaccination may not be an appropriate biomarker for vaccine efficacy.

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Data availability

All data relevant to the study are included in the article or uploaded as online supplementary information. Additional information regarding data may be obtained from the authors on reasonable request.

Abbreviations

BFA:

Brefeldin A

DC:

Dendritic cells

polyI:C:

Polyinosinic-polycytidylic acid

TLR:

Toll like receptor

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Acknowledgements

The authors would like to thank Dr. Alexandra Witter and Ms. Marissa Gonzales for assistance and discussions and the Beirne B. Carter Center for Immunology Research for use of the flow cytometry instruments.

Funding

This work was funded by National Institutes of Health grant R01CA166458 to Timothy N.J. Bullock. Aaron D. Stevens was supported by National Institutes of Health training Grant T32AI007496.

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

    Aaron D. Stevens & Timothy N. J. Bullock

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  1. Aaron D. Stevens
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Contributions

All authors conceived the project, designed experiments, interpreted data, and wrote the manuscript. A.D.S. performed experiments and analyzed data. T.N.J.B. supervised and acquired funding.

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Correspondence to Timothy N. J. Bullock.

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All mice were treated in accordance with policies established by the University of Virginia Animal Care and Use Committee (protocol 3292).

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Stevens, A.D., Bullock, T.N.J. Therapeutic vaccination targeting CD40 and TLR3 controls melanoma growth through existing intratumoral CD8 T cells without new T cell infiltration. Cancer Immunol Immunother 70, 2139–2150 (2021). https://doi.org/10.1007/s00262-020-02841-z

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