Abstract
The ideal tumor antigen for the development of a cancer immunotherapy is one that is expressed only in tumor cells. The epidermal growth factor receptor pathway substrate 8 gene (Eps8) might be an effective antigen for cancer immunotherapy as it is overexpressed in a variety of cancer cells but not in normal tissues. In this study, the potential utility of an Eps8-derived immunotherapy was tested in vitro and in vivo. Three computer-based algorithms were used to design eight Eps8 native epitopes with potentially high binding affinity to the HLA-A2.1 molecule, which is found at a high frequency in the Chinese population. Of these eight, three peptides with a moderate affinity to the HLA-A2.1 molecule were modified at anchor residue positions to achieve stronger immunogenicity. These four modified peptides displayed stronger binding affinity to HLA-A2.1 molecules on T2 cells and a lower dissociation rate. In functional assays with human PBMCs in vitro and in HLA-A2.1/Kb transgenic mice in vivo, CTLs primed by each native and modified peptide secreted IFN-γ and were toxic to cancer cells from a variety of tissue types in an HLA-A2.1-restricted and Eps8-specific manner. p101–109-2L and p276–284-1Y9V were superior to other modified and native epitopes both in vitro and in vivo. These results indicate that employing the native and modified epitopes identified here in Eps8-based immunotherapy for HLA-A2.1 positive cancer patients may result in efficient anticancer immune responses for diverse tumor types.
Similar content being viewed by others
Abbreviations
- APC:
-
Antigen-presenting cells
- APL:
-
Altered peptide ligand
- CTL:
-
Cytotoxic T lymphocyte
- DC:
-
Dendritic cell
- DC50 :
-
Dissociation complex 50
- Eps8:
-
Epidermal growth factor receptor pathway substrate 8 gene
- EGFR:
-
Epidermal growth factor receptor
- ELISPOT:
-
Enzyme-linked immunospot
- E/T:
-
Effector/target
- FI:
-
Fluorescence index
- IFN-γ:
-
Interferon-gamma
- IMP:
-
Influenza matrix protein
- LDH:
-
Lactate dehydrogenase
- MFI:
-
Mean fluorescence intensity
- PADRE:
-
Pan DR epitope peptides
- PBMC:
-
Peripheral blood mononuclear cell
- TAA:
-
Tumor-associated antigen
- TAP:
-
Transporter-associated protein
References
Gilboa E (2004) The promise of cancer vaccines. Nat Rev Cancer 4(5):401–411
Mocellin S, Semenzato G, Mandruzzato S, Riccardo RC (2004) Part II: vaccines for haematological malignant disorders. Lancet Oncol 5(12):727–737
Vigneron N, Stroobant V, Van den Eynde BJ, van der Bruggen P (2013) Database of T cell-defined human tumor antigens: the 2013 update. Cancer Immun 13:15
Anderson LJ, Cook DR, Yamamoto TN, Berger C, Maloney DG, Riddell SR (2011) Identification of MAGE-C1 (CT-7) epitopes for T-cell therapy of multiple myeloma. Cancer Immunol Immunother 60(7):985–997
Sun Z, Lethe B, Zhang Y, Russo V, Colau D, Stroobant V, Boon T, van der Bruggen P (2006) A new LAGE-1 peptide recognized by cytolytic T lymphocytes on HLA-A68 tumors. Cancer Immunol Immunother 55(6):644–652
Matsuzaki J, Qian F, Luescher I, Lele S, Ritter G, Shrikant PA, Gnjatic S, Old LJ, Odunsi K (2008) Recognition of naturally processed and ovarian cancer reactive CD8 + T cell epitopes within a promiscuous HLA class II T-helper region of NY-ESO-1. Cancer Immunol Immunother 57(8):1185–1195
Keilholz U, Letsch A, Busse A, Asemissen AM, Bauer S, Blau IW, Hofmann WK, Uharek L, Thiel E, Scheibenbogen C (2009) A clinical and immunologic phase 2 trial of Wilms tumor gene product 1 (WT1) peptide vaccination in patients with AML and MDS. Blood 113(26):6541–6548
Emens LA (2012) Re-purposing cancer therapeutics for breast cancer immunotherapy. Cancer Immunol Immunother 61(8):1299–1305
Parmiani G, Russo V, Maccalli C, Parolini D, Rizzo N, Maio M. (2014) Peptide-based vaccines for cancer therapy. Hum Vaccin Immunother, 10(11). Advance online publication. doi:10.4161/hv.29418
Li YH, Xue TY, He YZ, Du JW (2013) Novel oncoprotein EPS8: a new target for anticancer therapy. Future Oncol 9:1587–1594
Fazioli F, Minichiello L, Matoska V, Castagnino P, Miki T, Wong WT, Di Fiore PP (1993) Eps8, a substrate for the epidermal growth factor receptor kinase, enhances EGF-dependent mitogenic signals. EMBO J 12(10):3799–3808
Scita G, Nordstrom J, Carbone R, Tenca P, Giardina G, Gutkind S, Bjarnegard M, Betsholtz C, Di Fiore PP (1999) EPS8 and E3B1 transduce signals from Ras to Rac. Nature 401(6750):290–293
Scita G, Tenca P, Areces LB, Tocchetti A, Frittoli E, Giardina G, Ponzanelli I, Sini P, Innocenti M, Di Fiore PP (2001) An effector region in Eps8 is responsible for the activation of the Rac-specific GEF activity of Sos-1 and for the proper localization of the Rac-based actin-polymerizing machine. J Cell Biol 154(5):1031–1044
Disanza A, Carlier MF, Stradal TE, Didry D, Frittoli E, Confalonieri S, Croce A, Wehland J, Di Fiore PP, Scita G (2004) Eps8 controls actin-based motility by capping the barbed ends of actin filaments. Nat Cell Biol 6(12):1180–1188
Shieh DC, Lin DT, Yang BS, Kuan HL, Kao KJ (1996) High frequency of HLA-A*0207 subtype in Chinese population. Transfusion 36(9):818–821
Ruppert J, Sidney J, Celis E, Kubo RT, Grey HM, Sette A (1993) Prominent role of secondary anchor residues in peptide binding to HLA-A2.1 molecules. Cell 74(5):929–937
Tourdot S, Scardino A, Saloustrou E, Gross DA, Pascolo S, Cordopatis P, Lemonnier FA, Kosmatopoulos K (2000) A general strategy to enhance immunogenicity of low-affinity HLA-A2. 1-associated peptides: implication in the identification of cryptic tumor epitopes. Eur J Immunol 30(12):3411–3421
Kreher CR, Dittrich MT, Guerkov R, Boehm BO, Tary-Lehmann M (2003) CD4 + and CD8 + cells in cryopreserved human PBMC maintain full functionality in cytokine ELISPOT assays. J Immunol Methods 278(1–2):79–93
Rammensee H, Bachmann J, Emmerich NP, Bachor OA, Stevanovic S (1999) SYFPEITHI: database for MHC ligands and peptide motifs. Immunogenetics 50(3–4):213–219
Parker KC, Bednarek MA, Coligan JE (1994) Scheme for ranking potential HLA-A2 binding peptides based on independent binding of individual peptide side-chains. J Immunol 152(1):163–175
Nielsen M, Lundegaard C, Worning P, Lauemoller SL, Lamberth K, Buus S, Brunak S, Lund O (2003) Reliable prediction of T-cell epitopes using neural networks with novel sequence representations. Protein Sci 12(5):1007–1017
Greiner J, Li L, Ringhoffer M, Barth TF, Giannopoulos K, Guillaume P, Ritter G, Wiesneth M, Dohner H, Schmitt M (2005) Identification and characterization of epitopes of the receptor for hyaluronic acid-mediated motility (RHAMM/CD168) recognized by CD8 + T cells of HLA-A2-positive patients with acute myeloid leukemia. Blood 106(3):938–945
Passoni L, Scardino A, Bertazzoli C, Gallo B, Coluccia AM, Lemonnier FA, Kosmatopoulos K, Gambacorti-Passerini C (2002) ALK as a novel lymphoma -associated tumor antigen: identification of 2 HLA-A2.1-restricted CD8 + T-cell epitopes. Blood 99(6):2100–2106
Ohminami H, Yasukawa M, Fujita S (2000) HLA class I-restricted lysis of leukemia cells by a CD8(+) cytotoxic T-lymphocyte clone specific for WT1 peptide. Blood 95(1):286–293
Greiner J, Li L, Ringhoffer M, Barth TF, Giannopoulos K, Guillaume P, Ritter G, Wiesneth M, Dohner H, Schmitt M (2005) Identification and characterization of epitopes of the receptor for hyaluronic acid-mediated motility (RHAMM/CD168) recognized by CD8 + T cells of HLA-A2-positive patients with acute myeloid leukemia. Blood 106(3):938–945
Olson BM, Frye TP, Johnson LE, Fong L, Knutson KL, Disis ML, McNeel DG (2010) HLA-A2-restricted T-cell epitopes specific for prostatic acid phosphatase. Cancer Immunol Immunother 59(6):943–953
Alexander J, Sidney J, Southwood S, Ruppert J, Oseroff C, Maewal A, Snoke K, Serra HM, Kubo RT, Sette A et al (1994) Development of high potency universal DR-restricted helper epitopes by modification of high affinity DR-blocking peptides. Immunity 1:751–761
Li F, Yang D, Wang Y, Liu B, Deng Y, Wang L, Shang X, Tong W, Ni B, Wu Y (2009) Identification and modification of an HLA-A*0201-restricted cytotoxic T lymphocyte epitope from Ran antigen. Cancer Immunol Immunother 58:2039–2049
Parkhurst MR, Salgaller ML, Southwood S, Robbins PF, Sette A, Rosenberg SA, Kawakami Y (1996) Improved induction of melanoma-reactive CTL with peptides from the melanoma antigen gp100 modified at HLA-A*0201-binding residues. J Immunol 157(6):2539–2548
Valmori D, Fonteneau JF, Lizana CM, Gervois N, Lienard D, Rimoldi D, Jongeneel V, Jotereau F, Cerottini JC, Romero P (1998) Enhanced generation of specific tumor-reactive CTL in vitro by selected Melan-A/MART-1 immunodominant peptide analogues. J Immunol 160(4):1750–1758
Buhrman JD, Slansky JE (2013) Improving T cell responses to modified peptides in tumor vaccines. Immunol Res 55(1–3):34–47
Pinilla-Ibarz J, May RJ, Korontsvit T, Gomez M, Kappel B, Zakhaleva V, Zhang RH, Scheinberg DA (2006) Improved human T-cell responses against synthetic HLA-0201 analog peptides derived from the WT1 oncoprotein. Leukemia 20(11):2025–2033
Stuge TB, Holmes SP, Saharan S, Tuettenberg A, Roederer M, Weber JS, Lee PP (2004) Diversity and recognition efficiency of T cell responses to cancer. PLoS Med 1(2):e28
Wieckowski S, Baumgaertner P, Corthesy P, Voelter V, Romero P, Speiser DE, Rufer N (2009) Fine structural variations of alphabetaTCRs selected by vaccination with naturalversus altered self-antigen in melanoma patients. J Immunol 183(8):5397–5406
Le Gal FA, Ayyoub M, Dutoit V, Widmer V, Jager E, Cerottini JC, Dietrich PY, Valmori D (2005) Distinct structural TCR repertoires in naturally occurring versus vaccine-induced CD8 + T-cell responses to the tumor-specific antigen NY-ESO-1. J Immunother 28(3):252–257
Speiser DE, Baumgaertner P, Voelter V, Devevre E, Barbey C, Rufer N, Romero P (2008) Unmodified self antigen triggers human CD8 T cells with stronger tumor reactivity than altered antigen. Proc Natl Acad Sci USA 105(10):3849–3854
Acknowledgments
This work was supported by the National Natural Science Foundation of China (Grants 81372249 and 81300431), the Foundation of the Ministry of Education of China for Returned Scholars, the Research Fund for the Doctoral Program of Higher Education of the Ministry of National Education, China (Grant 20114433110012), the Project of Department of Education of Guangdong Province (Grant 2012KJCX0025), the Key Project of Science and Technology of Guangzhou City (Grant 12C22121595), and the Natural Science Foundation of Guangdong Province, China (Grant S2013040014449).
Conflict of interest
The authors declare that they have no conflicts of interest.
Author information
Authors and Affiliations
Corresponding authors
Additional information
Yuhua Li and Weijun Zhou have contributed to the work equally and should be regarded as co-first authors.
Rights and permissions
About this article
Cite this article
Li, Y., Zhou, W., Du, J. et al. Generation of cytotoxic T lymphocytes specific for native or modified peptides derived from the epidermal growth factor receptor pathway substrate 8 antigen. Cancer Immunol Immunother 64, 259–269 (2015). https://doi.org/10.1007/s00262-014-1631-y
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00262-014-1631-y