Healthy Donors Harbor Memory T Cell Responses to RAS Neo-Antigens
Abstract
:Simple Summary
Abstract
1. Introduction
2. Results
2.1. Initial Screening Against RAS-Mutant Crude Epitopes Reveal Strong and Frequent Responses to Several RAS-Mutant Neo-Antigens
2.2. T Cell Responses to Mutant RAS Protein Are Not Explained by Either Peptide Impurities or Cross-Reactivity with the Wild-Type Epitope
2.3. T Cells from Healthy Donors Demonstrate a Strong Response to Stimulation with Several Different Mutant RAS Epitopes
2.4. T Cells from Healthy Donors Respond to Stimulation with Mutant RAS Epitopes in an Ex Vivo Setting
2.5. T-Cell Clones Reactive to Mutant RAS Epitopes May Express Some Cross-Reactivity to Wt RAS Epitopes
2.6. RAS-Mutant-Specific T Cells in Healthy Donors Are Antigen-Experienced Memory T Cells
3. Discussion
4. Materials and Methods
4.1. Donors
4.2. Peptides
4.3. Enzyme-Linked Immunospot Assay
4.4. Intracellular Cytokine Staining
4.5. Establishment of T Cell Cultures Specific to Mutant RAS Epitopes
4.6. CD4+CD45RO+ Memory T Cell Cultures
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
References
- Dunn, G.P.; Old, L.J.; Schreiber, R.D. The Three Es of Cancer Immunoediting. Annu. Rev. Immunol. 2004, 22, 329–360. [Google Scholar] [CrossRef] [PubMed]
- Dunn, G.P.; Bruce, A.T.; Ikeda, H.; Old, L.J.; Schreiber, R.D. Cancer immunoediting: From immuno- surveillance to tumor escape. Nat. Immunol. 2002, 3, 991–998. [Google Scholar] [CrossRef] [PubMed]
- Posthuma, E.F.; Falkenburg, J.H.; Apperley, J.F.; Gratwohl, A.; Roosnek, E.; Hertenstein, B.; Schipper, R.F.; Schreuder, G.; D’Amaro, J.; Oudshoorn, M.; et al. HLA-B8 and HLA-A3 coexpressed with HLA-B8 are associated with a reduced risk of the development of chronic myeloid leukemia. The Chronic Leukemia Working Party of the EBMT. Blood 1999, 93, 3863–3865. [Google Scholar]
- Kuželová, K.; Brodská, B.; Fuchs, O.; Dobrovolná, M.; Soukup, P.; Cetkovský, P. Altered HLA Class I Profile Associated with Type A/D Nucleophosmin Mutation Points to Possible Anti-Nucleophosmin Immune Response in Acute Myeloid Leukemia. PLoS ONE 2015, 10, e0127637. [Google Scholar] [CrossRef] [PubMed]
- Schumacher, T.N.; Schreiber, R.D. Neoantigens in cancer immunotherapy. Science 2015, 348, 69–74. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Greiner, J.; Schneider, V.; Schmitt, M.; Götz, M.; Döhner, K.; Wiesneth, M.; Döhner, H.; Hofmann, S. Immune responses against the mutated region of cytoplasmatic NPM1 might contribute to the favorable clinical outcome of AML patients with NPM1 mutations (NPM1mut). Blood 2013, 122, 1087–1088. [Google Scholar] [CrossRef]
- Coulie, P.; Brichard, V.; Van Pel, A.; Wolfel, T.; Schneider, J.; Traversari, C.; Mattei, S.; De Plaen, E.; Lurquin, C.; Szikora, J.-P.; et al. A new gene coding for a differentiation antigen recognized by autologous cytolytic T lymphocytes on HLA-A2 melanomas. J. Exp. Med. 1994, 180, 35–42. [Google Scholar] [CrossRef]
- Pittet, M.J.; Valmori, D.; Dunbar, P.R.; Speiser, D.E.; Liénard, D.; Lejeune, F.; Fleischhauer, K.; Cerundolo, V.; Cerottini, J.-C.; Romero, P. High Frequencies of Naive Melan-a/Mart-1–Specific Cd8+ T Cells in a Large Proportion of Human Histocompatibility Leukocyte Antigen (Hla)-A2 Individuals. J. Exp. Med. 1999, 190, 705–716. [Google Scholar] [CrossRef] [Green Version]
- Nangalia, J.; Massie, C.; Baxter, E.; Nice, F.; Gundem, G.; Wedge, D.; Avezov, E.; Li, J.; Kollmann, K.; Kent, D.; et al. Somatic CALRMutations in Myeloproliferative Neoplasms with Nonmutated JAK2. N. Engl. J. Med. 2013, 369, 2391–2405. [Google Scholar] [CrossRef] [Green Version]
- Klampfl, T.; Gisslinger, H.; Harutyunyan, A.S.; Nivarthi, H.; Rumi, E.; Milosevic, J.D.; Them, N.C.C.; Berg, T.; Gisslinger, B.; Pietra, D.; et al. Somatic Mutations of Calreticulin in Myeloproliferative Neoplasms. N. Engl. J. Med. 2013, 369, 2379–2390. [Google Scholar] [CrossRef] [Green Version]
- Holmström, M.O.; Riley, C.H.; Svane, I.M.; Hasselbalch, H.C.; Andersen, M.H. The CALR exon 9 mutations are shared neoantigens in patients with CALR mutant chronic myeloproliferative neoplasms. Leukemia 2016, 30, 2413–2416. [Google Scholar] [CrossRef] [PubMed]
- Holmström, M.O.; Martinenaite, E.; Ahmad, S.M.; Met, Ö.; Friese, C.; Kjær, L.; Riley, C.H.; Straten, P.T.; Svane, I.M.; Hasselbalch, H.C.; et al. The calreticulin (CALR) exon 9 mutations are promising targets for cancer immune therapy. Leukemia 2017, 32, 429–437. [Google Scholar] [CrossRef] [PubMed]
- Holmström, M.O.; Ahmad, S.M.; Klausen, U.; Bendtsen, S.K.; Martinenaite, E.; Riley, C.H.; Svane, I.M.; Kjær, L.; Skov, V.; Ellervik, C.; et al. High frequencies of circulating memory T cells specific for calreticulin exon 9 mutations in healthy individuals. Blood Cancer J. 2019, 9, 8. [Google Scholar] [CrossRef]
- Prior, I.A.; Lewis, K.E.; Mattos, C. A Comprehensive Survey of Ras Mutations in Cancer. Cancer Res. 2012, 72, 2457–2467. [Google Scholar] [CrossRef] [Green Version]
- Shono, Y.; Tanimura, H.; Iwahashi, M.; Tsunoda, T.; Tani, M.; Tanaka, H.; Matsuda, K.; Yamaue, H. Specific T-cell immunity against Ki-ras peptides in patients with pancreatic and colorectal cancers. Br. J. Cancer 2003, 88, 530–536. [Google Scholar] [CrossRef] [Green Version]
- Kubuschok, B.; Neumann, F.; Breit, R.; Sester, M.; Schormann, C.; Wagner, C.; Hartmann, F.; Remberger, K.; Schilling, M.; Pfreundschuh, M. Naturally Occurring T-Cell Response against Mutated p21 Ras Oncoprotein in Pancreatic Cancer. Clin. Cancer Res. 2006, 12, 1365–1372. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Qin, H.; Chen, W.; Takahashi, M.; Disis, M.L.; Byrd, D.R.; McCahill, L.; Bertram, K.A.; Fenton, R.G.; Peace, D.J.; A Cheever, M. CD4+ T-cell immunity to mutated ras protein in pancreatic and colon cancer patients. Cancer Res. 1995, 55, 2984–2987. [Google Scholar] [PubMed]
- Gedde-Dahl, T.; Eriksen, J.A.; Thorsby, E.; Gaudernack, G. T-cell responses against products of oncogenes: Generation and characterization of human T-cell clones specific for p21 ras-derived synthetic peptides. Hum. Immunol. 1992, 33, 266–274. [Google Scholar] [CrossRef]
- Gjertsen, M.; Breivik, J.; Saeterdal, I.; Thorsby, E.; Gaudernack, G.; Bakka, A.; Reide, O.S.O.; Solheim, B. Vaccination with mutant ras peptides and induction of T-cell responsiveness in pancreatic carcinoma patients carrying the corresponding RAS mutation. Lancet 1995, 346, 1399–1400. [Google Scholar] [CrossRef]
- Gjertsen, M.K.; Bakka, A.; Breivik, J.; Saeterdal, I.; Gedde-Dahl, T.; Stokke, K.T.; Solheim, B.G.; Egge, T.S.; Søreide, O.; Thorsby, E.; et al. Ex vivo ras peptide vaccination in patients with advanced pancreatic cancer: Results of a phase I/II study. Int. J. Cancer 1996, 65, 450–453. [Google Scholar] [CrossRef]
- Rahma, O.E.; Hamilton, J.M.; Wojtowicz, M.; Dakheel, O.; Bernstein, S.; Liewehr, D.J.; Steinberg, S.M.; Khleif, S.N. The immunological and clinical effects of mutated ras peptide vaccine in combination with IL-2, GM-CSF, or both in patients with solid tumors. J. Transl. Med. 2014, 12, 55. [Google Scholar] [CrossRef] [Green Version]
- Toubaji, A.; Achtar, M.; Provenzano, M.; Herrin, V.E.; Behrens, R.; Hamilton, M.; Bernstein, S.; Venzon, D.; Gause, B.; Marincola, F.; et al. Pilot study of mutant ras peptide-based vaccine as an adjuvant treatment in pancreatic and colorectal cancers. Cancer Immunol. Immunother. 2008, 57, 1413–1420. [Google Scholar] [CrossRef] [PubMed]
- Abrams, S.I.; Khleif, S.N.; Bergmann-Leitner, E.S.; Kantor, J.A.; Chung, Y.; Hamilton, J.; Schlom, J. Generation of Stable CD4+and CD8+T Cell Lines from Patients Immunized withrasOncogene-Derived Peptides Reflecting Codon 12 Mutations. Cell. Immunol. 1997, 182, 137–151. [Google Scholar] [CrossRef]
- Gjertsen, M.; Sæterdal, I.; Sæbøe-Larssen, S.; Gaudernack, G. HLA-A3 restricted mutant ras specific cytotoxic T-lymphocytes induced by vaccination with T-helper epitopes. J. Mol. Med. 2003, 81, 43–50. [Google Scholar] [CrossRef]
- Jung, S. Human T lymphocytes recognize a peptide of single point-mutated, oncogenic ras proteins. Trends Genet. 1991, 7, 78. [Google Scholar] [CrossRef]
- Fossum, B.; Olsen, A.C.; Thorsby, E.; Gaudernack, G. CD8+ T cells from a patient with colon carcinoma, specific for a mutant p21-Ras-derived peptide (Gly13-->Asp), are cytotoxic towards a carcinoma cell line harbouring the same mutation. Cancer Immunol. Immunother. 1995, 40, 165–172. [Google Scholar]
- Gjertsen, M.K.; Bjørheim, J.; Saetardal, I.; Myklebust, J.; Gaudernack, G. Cytotoxic CD4 + and CD8 + T lymphocytes, generated by mutant p21-ras (12Val) peptide vaccination of a patient, recognize 12Val-dependent nested epitopes present within the vaccine peptide and kill autologous tumour cells carrying this mutation. Int. J. Cancer 1997, 72, 784–790. [Google Scholar] [CrossRef]
- Moodie, Z.; Price, L.; Gouttefangeas, C.; Mander, A.; Janetzki, S.; Löwer, M.; Welters, M.J.P.; Ottensmeier, C.H.; Van Der Burg, S.H.; Britten, C.M. Response definition criteria for ELISPOT assays revisited. Cancer Immunol. Immunother. 2010, 59, 1489–1501. [Google Scholar] [CrossRef] [Green Version]
- Prior, I.A.; Hood, F.E.; Hartley, J.L. The Frequency of Ras Mutations in Cancer. Cancer Res. 2020, 80, 2969–2974. [Google Scholar] [CrossRef] [Green Version]
- Wedén, S.; Klemp, M.; Gladhaug, I.P.; Møller, M.; Eriksen, J.A.; Gaudernack, G.; Buanes, T. Long-term follow-up of patients with resected pancreatic cancer following vaccination against mutant K-ras. Int. J. Cancer 2010, 128, 1120–1128. [Google Scholar] [CrossRef]
- Almoguera, C.; Shibata, D.; Forrester, K.; Martin, J.; Arnheim, N.; Perucho, M. Most human carcinomas of the exocrine pancreas contain mutant c-K-ras genes. Cell 1988, 53, 549–554. [Google Scholar] [CrossRef] [Green Version]
- Mahnke, Y.D.; Brodie, T.M.; Sallusto, F.; Roederer, M.; Lugli, E. The who’s who of T-cell differentiation: Human memory T-cell subsets. Eur. J. Immunol. 2013, 43, 2797–2809. [Google Scholar] [CrossRef]
- Fossum, B.; Gedde-Dahl, T., III; Breivik, J.; Eriksen, J.A.; Spurkland, A.; Thorsby, E.; Gaudernack, G. p21-ras-peptide-specific T-cell responses in a patient with colorectal cancer. CD4+ and CD8+ T cells recognize a peptide corresponding to a common mutation (13Gly → Asp). Int. J. Cancer 1994, 56, 40–45. [Google Scholar] [CrossRef] [PubMed]
- Gjertsen, M.; Buanes, T.; Rosseland, A.R.; Bakka, A.; Gladhaug, I.; Eriksen, J.A.; Lothe, R.A.; Gaudernack, G. Intradermal ras peptide vaccination with granulocyte-macrophage colony-stimulating factor as adjuvant: Clinical and immunological responses in patients with pancreatic adenocarcinoma. Int. J. Cancer 2001, 92, 441–450. [Google Scholar] [CrossRef]
- Keilholz, U.; Weber, J.S.; Finke, J.H.; Gabrilovich, D.I.; Kast, W.M.; Disis, M.L.; Kirkwood, J.M.; Scheibenbogen, C.; Schlom, J.; Maino, V.C.; et al. Immunologic Monitoring of Cancer Vaccine Therapy: Results of a Workshop Sponsored by the Society for Biological Therapy. J. Immunother. 2002, 25, 97–138. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Gjertsen, M.; Gaudernack, G. Mutated Ras Peptides as Vaccines in Immunotherapy of Cancer. Vox Sang. 1998, 74, 489–495. [Google Scholar] [CrossRef]
- 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. 2010, 10, 554–567. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Falini, B.; Mecucci, C.; Tiacci, E.; Alcalay, M.; Rosati, R.; Pasqualucci, L.; La Starza, R.; Diverio, D.; Colombo, E.; Santucci, A.; et al. Cytoplasmic Nucleophosmin in Acute Myelogenous Leukemia with a Normal Karyotype. N. Engl. J. Med. 2005, 352, 254–266. [Google Scholar] [CrossRef]
- Cobbold, M.; De La Peña, H.; Norris, A.; Polefrone, J.M.; Qian, J.; English, A.M.; Cummings, K.L.; Penny, S.; Turner, J.E.; Cottine, J.; et al. MHC Class I-Associated Phosphopeptides Are the Targets of Memory-like Immunity in Leukemia. Sci. Transl. Med. 2013, 5, 203ra125. [Google Scholar] [CrossRef] [Green Version]
- Yu, W.; Jiang, N.; Ebert, P.J.R.; Kidd, B.A.; Müller, S.; Lund, P.J.; Juang, J.; Adachi, K.; Tse, T.; Birnbaum, M.E.; et al. Clonal Deletion Prunes but Does Not Eliminate Self-Specific αβ CD8+ T Lymphocytes. Immunity 2015, 42, 929–941. [Google Scholar] [CrossRef] [Green Version]
- Kim, J.Y.; Hong, S.-M. Precursor Lesions of Pancreatic Cancer. Oncol. Res. Treat. 2018, 41, 603–610. [Google Scholar] [CrossRef] [PubMed]
- Retsinformation. Available online: https://www.retsinformation.dk/eli/lta/2017/1083 (accessed on 16 September 2020).
CODON 12 PEPTIDES | Amino Acid Sequence |
---|---|
G12A: | EYKLVVVGAAGVGKSALTI |
G12C: | EYKLVVVGACGVGKSALTI |
G12D: | EYKLVVVGADGVGKSALTI |
G12R: | EYKLVVVGARGVGKSALTI |
G12S: | EYKLVVVGASGVGKSALTI |
G12V: | EYKLVVVGAVGVGKSALTI |
G12wt: | EYKLVVVGAGGVGKSALTI |
CODON 13 PEPTIDES | |
G13A: | YKLVVVGAGAVGKSALTIQ |
G13C: | YKLVVVGAGCVGKSALTIQ |
G13D: | YKLVVVGAGDVGKSALTIQ |
G13R: | YKLVVVGAGRVGKSALTIQ |
G13S: | YKLVVVGAGSVGKSALTIQ |
G13V: | YKLVVVGAGVVGKSALTIQ |
G13wt: | YKLVVVGAGGVGKSALTIQ |
CODON 61 PEPTIDES | |
Q61E: | LLDILDTAGEEEYSAMRDQ |
Q61H: | LLDILDTAGHEEYSAMRDQ |
Q61K: | LLDILDTAGKEEYSAMRDQ |
Q61L: | LLDILDTAGLEEYSAMRDQ |
Q61P: | LLDILDTAGPEEYSAMRDQ |
Q61R: | LLDILDTAGREEYSAMRDQ |
Publisher’s Note: MDPI stays neutral with regard to jurisdictional claims in published maps and institutional affiliations. |
© 2020 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
Share and Cite
Holmström, M.O.; Andersen, M.H. Healthy Donors Harbor Memory T Cell Responses to RAS Neo-Antigens. Cancers 2020, 12, 3045. https://doi.org/10.3390/cancers12103045
Holmström MO, Andersen MH. Healthy Donors Harbor Memory T Cell Responses to RAS Neo-Antigens. Cancers. 2020; 12(10):3045. https://doi.org/10.3390/cancers12103045
Chicago/Turabian StyleHolmström, Morten Orebo, and Mads Hald Andersen. 2020. "Healthy Donors Harbor Memory T Cell Responses to RAS Neo-Antigens" Cancers 12, no. 10: 3045. https://doi.org/10.3390/cancers12103045