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.
This is a preview of subscription content, access via your institution
Open Access articles citing this article.
Tumor microenvironment antigens
Seminars in Immunopathology Open Access 29 September 2022
Subscribe to this journal
Receive 12 digital issues and online access to articles
$119.00 per year
only $9.92 per issue
Rent or buy this article
Get just this article for as long as you need it
Prices may be subject to local taxes which are calculated during checkout
Mullen, A. C. & Wrana, J. L. TGF-β family signaling in embryonic and somatic stem-cell renewal and differentiation. Cold Spring Harb. Perspect. Biol. 9, a027987 (2017).
Batlle, E. & Massagué, J. Transforming grown factor-β signaling in immunity and cancer. Immunity 50, 924–940 (2019).
Flavell, R. A., Sanjabi, S., Wrzesinski, S. H. & Licona-Limón, P. The polarization of immune cells in the tumour environment by TGFβ. Nat. Rev. Immunol. 10, 554–567 (2010).
Massagué, J. TGFβ in cancer. Cell 134, 215–230 (2008).
Elliott, R. L. & Blobe, G. C. Role of transforming growth factor beta in human cancer. J. Clin. Oncol. 23, 2078–2093 (2005).
Hao, Y., Baker, D. & Dijke, P. T. TGF-β-mediated epithelial-mesenchymal transition and cancer metastasis. Int. J. Mol. Sci. 20, 2767 (2019).
Kobayashi, H. et al. Cancer-associated fibroblasts in gastrointestinal cancer. Nat. Rev. Gastroenterol. Hepatol. 16, 282–295 (2019).
Chen, X. & Song, E. Turning foes to friends: targeting cancer-associated fibroblasts. Nat. Rev. Drug Discov. 18, 99–115 (2019).
Andersen, M. H. Anti-regulatory T cells. Semin. Immunopathol. 39, 317–326 (2017).
Munir, S. et al. HLA-restricted CTL that are specific for the immune checkpoint ligand PD-L1 occur with high frequency in cancer patients. Cancer Res. 73, 1764–1776 (2013).
Ahmad, S. M. et al. The inhibitory checkpoint, PD-L2, is a target for effector T cells: Novel possibilities for immune therapy. Oncoimmunology https://doi.org/10.1080/2162402X.2017.1390641 (2017).
Sørensen, R. B. et al. Spontaneous cytotoxic T-cell reactivity against indoleamine 2,3-dioxygenase-2. Cancer Res. 71, 2038–2044 (2011).
Martinenaite, E. et al. Frequent adaptive immune responses against arginase-1. Oncoimmunology 7, e1404215 (2017).
Weis-Banke, S. E. et al. The metabolic enzyme arginase-2 is a potential target for novel immune modulatory vaccines. Oncoimmunology 9, 1771142 (2020).
Andersen, M. H. The T-win® technology: immune-modulating vaccines. Semin. Immunopathol. 41, 87–95 (2019).
Iversen, T. Z. et al. Long-lasting disease stabilization in the absence of toxicity in metastatic lung cancer patients vaccinated with an epitope derived from indoleamine 2,3 dioxygenase. Clin. Cancer Res. 20, 221–232 (2014).
Svane, I.-M., Kjeldsen, J. W., Lorentzen, C. L., Martinenaite, E. & Andersen, M. H. Clinical efficacy and immunity of combination therapy with nivolumab and IDO/PD-L1 peptide vaccine in patients with metastatic melanoma: a phase I/II trial. Ann. Oncol. 31, S1176 (2020).
Moodie, Z. et al. Response definition criteria for ELISPOT assays revisited. Cancer Immunol. Immunother. 59, 1489–1501 (2010).
Holmstrom, M. O. et al. The calreticulin (CALR) exon 9 mutations are promising targets for cancer immune therapy. Leukemia 32, 429–437 (2018).
Andersen, M. H. et al. Phosphorylated peptides can be transported by TAP molecules, presented by class I MHC molecules, and recognized by phosphopeptide-specific CTL. J. Immunol. 163, 3812–3818 (1999).
Schneider, C. A., Rasband, W. S. & Eliceiri, K. W. NIH Image to ImageJ: 25 years of image analysis. Nat. Methods 9, 671–675 (2012).
Martinenaite, E. et al. CCL22-specific T cells: modulating the immunosuppressive tumor microenvironment. Oncoimmunology 5, e1238541 (2016).
Met, Ö., Balslev, E., Flyger, H. & Svane, I. M. High immunogenic potential of p53 mRNA-transfected dendritic cells in patients with primary breast cancer. Breast Cancer Res. Treat. 125, 395–406 (2011).
Hjortsø, M. D. et al. Tryptophan 2,3-dioxygenase (TDO)-reactive T cells differ in their functional characteristics in health and cancer. Oncoimmunology 4, 968480 (2015).
Gnjatic, S. et al. Cross-presentation of HLA class I epitopes from exogenous NY-ESO-1 polypeptides by nonprofessional APCs. J. Immunol. 170, 1191–1196 (2003).
Clark, R. E. et al. Direct evidence that leukemic cells present HLA-associated immunogenic peptides derived from the BCR-ABL b3a2 fusion protein. Blood https://doi.org/10.1182/blood.V98.10.2887 (2001).
Wang, X. F. et al. The role of indoleamine 2,3-dioxygenase (IDO) in immune tolerance: Focus on macrophage polarization of THP-1 cells. Cell. Immunol. 289, 42–48 (2014).
Rammensee, H.-G., Bachmann, J., Emmerich, N. N., Bachor, O. A. & Stevanovic, S. SYFPEITHI: database for MHC ligands and peptide motifs. www.syfpeithi.de. Accessed 30 Aug 2019.
Thomas, D. A. & Massagué, J. TGF-β directly targets cytotoxic T cell functions during tumor evasion of immune surveillance. Cancer Cell 8, 369–380 (2005).
Rook, A. H. et al. Effects of transforming growth factor beta on the functions of natural killer cells: depressed cytolytic activity and blunting of interferon responsiveness. J. Immunol. 136, 3916–3920 (1986).
Torroella-Kouri, M. et al. Identification of a subpopulation of macrophages in mammary tumor-bearing mice that are neither M1 nor M2 and are less differentiated. Cancer Res. 69, 4800–4809 (2009).
Gabrilovich, D. I., Ostrand-Rosenberg, S. & Bronte, V. Coordinated regulation of myeloid cells by tumours. Nat. Rev. Immunol. 12, 253–268 (2012).
Pang, Y. et al. TGF-β Signaling in myeloid cells is required for tumor metastasis. Cancer Discov. 3, 936–951 (2013).
Yang, L. et al. Abrogation of TGFβ signaling in mammary carcinomas recruits Gr-1+CD11b+ myeloid cells that promote metastasis. Cancer Cell 13, 23–35 (2008).
Keilholz, U. et al. Immunologic monitoring of cancer vaccine therapy: results of a workshop sponsored by the Society for Biological Therapy. J. Immunother. 25, 97–138 (2002).
Rodon, J. et al. First-in-human dose study of the novel transforming growth factor-β receptor I kinase inhibitor LY2157299 monohydrate in patients with advanced cancer and glioma. Clin. Cancer Res. 21, 553–560 (2015).
Melisi, D. et al. Galunisertib plus gemcitabine vs. gemcitabine for first-line treatment of patients with unresectable pancreatic cancer. Br. J. Cancer 119, 1208–1214 (2018).
Giaccone, G. et al. A phase III study of belagenpumatucel-L, an allogeneic tumour cell vaccine, as maintenance therapy for non-small cell lung cancer. Eur. J. Cancer 51, 2321–2329 (2015).
Formenti, S. C. et al. Focal irradiation and systemic TGFb blockade in metastatic breast cancer. Clin. Cancer Res. 24, 2493–2504 (2018).
Mascarenhas, J. et al. Anti-transforming growth factor-β therapy in patients with myelofibrosis. Leuk. Lymphoma 55, 450–452 (2014).
Takaku, S. et al. Blockade of TGF-β enhances tumor vaccine efficacy mediated by CD8 + T cells. Int. J. Cancer 126, 1666–1674 (2010).
Terabe, M. et al. Synergistic enhancement of CD8+ T cell-mediated tumor vaccine efficacy by an anti-transforming growth factor-β monoclonal antibody. Clin. Cancer Res. 15, 6560–6569 (2009).
Ueda, R. et al. Systemic inhibition of transforming growth factor-β in glioma-bearing mice improves the therapeutic efficacy of glioma-associated antigen peptide vaccines. Clin. Cancer Res. 15, 6551–6559 (2009).
Ma, Y. et al. Targeting TGF-β1 by employing a vaccine ameliorates fibrosis in a mouse model of chronic colitis. Inflamm. Bowel Dis. 16, 1040–1050 (2010).
Chu, X. et al. Combined immunization against TGF-β1 enhances HPV16 E7-specific vaccine-elicited antitumour immunity in mice with grafted TC-1 tumours. Artif. Cells Nanomed. Biotechnol. 46, 1199–1209 (2018).
Ødum, N. Anti-regulatory T cells are natural regulatory effector T cells. Cell Stress 3, 310–311 (2019).
Munir Ahmad, S. et al. PD-L1 peptide co-stimulation increases immunogenicity of a dendritic cell-based cancer vaccine. Oncoimmunology 5, e1202391 (2016).
Rahma, O. E., Gammoh, E., Simon, R. M. & Khleif, S. N. Is the ‘3+3’ dose-escalation Phase i Clinical trial design suitable for therapeutic cancer vaccine development? A recommendation for alternative design. Clin. Cancer Res. 20, 4758–4767 (2014).
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.
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.
Rights and permissions
About this article
Cite this article
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
- Immune regulation
- T cells
- adaptive immunity
This article is cited by
Novel immune modulatory vaccines targeting TGFβ
Cellular & Molecular Immunology (2023)
Tumor microenvironment antigens
Seminars in Immunopathology (2023)
Characterization of TGFβ-specific CD4+T cells through the modulation of TGFβ expression in malignant myeloid cells
Cellular & Molecular Immunology (2021)