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Cytotoxic T cells isolated from healthy donors and cancer patients kill TGFβ-expressing cancer cells in a TGFβ-dependent manner

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Abstract

Transforming growth factor-beta (TGFβ) is a highly potent immunosuppressive cytokine. Although TGFβ is a tumor suppressor in early/premalignant cancer lesions, the cytokine has several tumor-promoting effects in advanced cancer; abrogation of the antitumor immune response is one of the most important tumor-promoting effects. As several immunoregulatory mechanisms have recently been shown to be targets of specific T cells, we hypothesized that TGFβ is targeted by naturally occurring specific T cells and thus could be a potential target for immunomodulatory cancer vaccination. Hence, we tested healthy donor and cancer patient T cells for spontaneous T-cell responses specifically targeting 38 20-mer epitopes derived from TGFβ1. We identified numerous CD4+ and CD8+ T-cell responses against several epitopes in TGFβ. Additionally, several ex vivo responses were identified. By enriching specific T cells from different donors, we produced highly specific cultures specific to several TGFβ-derived epitopes. Cytotoxic CD8+ T-cell clones specific for both a 20-mer epitope and a 9-mer HLA-A2 restricted killed epitope peptide were pulsed in HLA-A2+ target cells and killed the HLA-A2+ cancer cell lines THP-1 and UKE-1. Additionally, stimulation of THP-1 cancer cells with cytokines that increased TGFβ expression increased the fraction of killed cells. In conclusion, we have shown that healthy donors and cancer patients harbor CD4+ and CD8+ T cells specific for TGFβ-derived epitopes and that cytotoxic T cells with specificity toward TGFβ-derived epitopes are able to recognize and kill cancer cell lines in a TGFβ-dependent manner.

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Acknowledgements

This project was supported by the Danish Health Authority grant “Empowering Cancer Immunotherapy in Denmark”, grant number 4-1612-236/8, the Copenhagen University Hospital, Herlev, and Gentofte, and it was also supported through a research funding agreement between IO Biotech ApS and the National Center for Cancer Immune Therapy (CCIT-DK). We thank Anne Rahbech and Sara Ram Petersen for assistance to MOH in performing western blotting. We greatly appreciate the tremendous technical support from Merete Jonassen.

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M.O.H. performed experiments, interpreted the data, and wrote the manuscript, R.E.J.M. performed experiments and interpreted the data, M.A.P. performed experiments and interpreted the data, E.M. performed experiments and interpreted the data, S.E.W.-B. performed experiments and interpreted the data, M.A.-J. performed experiments and interpreted the data, S.K.B. performed experiments and interpreted the data, Ö.M. provided critical reagents, interpreted data, and wrote the manuscript, A.W.P. interpreted the data, M.D. interpreted the data, I.M.S. interpreted the data, and M.H.A. conceived the project, interpreted the data, and wrote the manuscript.

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Correspondence to Mads Hald Andersen.

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Competing interests

It should be noted, however, that Mads Hald Andersen has generated an invention based on the use of TGFβ for vaccinations. The rights to the invention have been transferred to Copenhagen University Hospital Herlev according to the Danish Law of Public Inventions at Public Research Institutions. The capital region has licensed the rights to the company IO Biotech ApS. The patent application was filed by IO Biotech ApS. Mads Hald Andersen is a board member, consultant, and shareholder in IO Biotech. Evelina Martinenaite and Ayako Wakatsuki Pedersen are employees at IO Biotech ApS. Inge Marie Svane is a consultant and shareholder in IO Biotech. The remaining authors declare no competing interests.

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Holmström, M.O., Mortensen, R.E.J., Pavlidis, A.M. et al. Cytotoxic T cells isolated from healthy donors and cancer patients kill TGFβ-expressing cancer cells in a TGFβ-dependent manner. Cell Mol Immunol 18, 415–426 (2021). https://doi.org/10.1038/s41423-020-00593-5

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