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Immunogenicity of the carcinoembryonic antigen derived peptide 694 in HLA-A2 healthy donors and colorectal carcinoma patients

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Abstract

Carcinoembryonic antigen (CEACAM5) is commonly overexpressed in human colon cancer. Several antigenic peptides recognized by cytolytic CD8+ T-cells have been identified and used in colon cancer phase-I vaccination clinical trials. The HLA-A*0201-binding CEA694–702 peptide was recently isolated from acid eluted MHC-I associated peptides from a human colon tumor cell line. However, the immunogenicity of this peptide in humans remains unknown. We found that the peptide CEA694–702 binds weakly to HLA-A*0201 molecules and is ineffective at inducing specific CD8+ T-cell responses in healthy donors. Immunogenic-altered peptide ligands with increased affinity for HLA-A*0201 were identified. Importantly, the elicited cytolytic T lymphocyte (CTL) lines and clones cross-reacted with the wild-type CEA694–702 peptide. Tumor cells expressing CEA were recognized in a peptide and HLA-A*0201 restricted fashion, but high-CEA expression levels appear to be required for CTL recognition. Finally, CEA-specific T-cell precursors could be readily expanded by in vitro stimulation of peripheral blood mononuclear cell (PBMC) from colon cancer patients with altered CEA peptide. However, the CEA-specific CD8+ T-cell clones derived from cancer patients revealed low-functional avidity and impaired tumor-cell recognition. Together, using T-cells to demonstrate the processing and presentation of the peptide CEA694-702, we were able to corroborate its presentation by tumor cells. However, the low avidity of the specific CTLs generated from cancer patients as well as the high-antigen expression levels required for CTL recognition pose serious concerns for the use of CEA694-702 in cancer immunotherapy.

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Abbreviations

h:

Hour

HLA-A2:

HLA-*0201

mAb:

Monoclonal antibody

MFI:

Mean fluorescence intensity

n.d.:

Not done

PCR:

Polymerase chain reaction

References

  1. Altman JD, Moss PA, Goulder PJ, Barouch DH, McHeyzer-Williams MG, Bell JI, McMichael AJ, Davis MM (1996) Phenotypic analysis of antigen-specific T lymphocytes. Science 274:94–96

    Article  PubMed  CAS  Google Scholar 

  2. Alves PM, Faure O, Graff-Dubois S, Cornet S, Bolonakis I, Gross DA, Miconnet I, Chouaib S, Fizazi K, Soria JC, Lemonnier FA, Kosmatopoulos K (2006) Steap, a prostate tumor antigen, is a target of human CD8(+) T cells. Cancer Immunol Immunother 55:1504–1514

    Article  CAS  Google Scholar 

  3. Alves PM, Faure O, Graff-Dubois S, Gross DA, Cornet S, Chouaib S, Miconnet I, Lemonnier FA, Kosmatopoulos K (2003) Epha2 as target of anticancer immunotherapy: identification of HLA-A*0201-restricted epitopes. Cancer Res 63:8476–8480

    PubMed  CAS  Google Scholar 

  4. Ayyoub M, Migliaccio M, Guillaume P, Lienard D, Cerottini JC, Romero P, Levy F, Speiser DE, Valmori D (2001) Lack of tumor recognition by Htert peptide 540–548-specific CD8(+) T cells from melanoma patients reveals inefficient antigen processing. Eur J Immunol 31:2642–2651

    Article  PubMed  CAS  Google Scholar 

  5. Berinstein NL (2002) Carcinoembryonic antigen as a target for therapeutic anticancer vaccines: a review. J Clin Oncol 20:2197–2207

    Article  PubMed  CAS  Google Scholar 

  6. Dalerba P, Maccalli C, Casati C, Castelli C, Parmiani G (2003) Immunology and immunotherapy of colorectal cancer. Crit Rev Oncol Hematol 46:33–57

    Article  PubMed  Google Scholar 

  7. Dupont J, Latouche JB, Ma C, Sadelain M (2005) Artificial antigen-presenting cells transduced with telomerase efficiently expand epitope-specific, human leukocyte antigen-restricted cytotoxic T cells. Cancer Res 65:5417–5427

    Article  PubMed  CAS  Google Scholar 

  8. Eidelman FJ, Fuks A, DeMarte L, Taheri M, Stanners CP (1993) Human carcinoembryonic antigen, an intercellular adhesion molecule, blocks fusion and differentiation of rat myoblasts. J Cell Biol 123:467–475

    Article  PubMed  CAS  Google Scholar 

  9. Faure O, Graff-Dubois S, Bretaudeau L, Derre L, Gross DA, Alves PM, Cornet S, Duffour MT, Chouaib S, Miconnet I, Gregoire M, Jotereau F, Lemonnier FA, Abastado JP, Kosmatopoulos K (2004) Inducible Hsp70 as target of anticancer immunotherapy: identification of HLA-A*0201-restricted epitopes. Int J Cancer 108:863–870

    Article  PubMed  CAS  Google Scholar 

  10. Fong L, Hou Y, Rivas A, Benike C, Yuen A, Fisher GA, Davis MM, Engleman EG (2001) Altered peptide ligand vaccination with Flt3 ligand expanded dendritic cells for tumor immunotherapy. Proc Natl Acad Sci USA 98:8809–8814

    Article  PubMed  CAS  Google Scholar 

  11. Goldberg AL, Cascio P, Saric T, Rock KL (2002) The importance of the proteasome and subsequent proteolytic steps in the generation of antigenic peptides. Mol Immunol 39:147–164

    Article  PubMed  CAS  Google Scholar 

  12. Graham RA, Wang S, Catalano PJ, Haller DG (1998) Postsurgical surveillance of colon cancer: preliminary cost analysis of physician examination, carcinoembryonic antigen testing, chest X-ray, and colonoscopy. Ann Surg 228:59–63

    Article  PubMed  CAS  Google Scholar 

  13. Hammarstrom S (1999) The carcinoembryonic antigen (Cea) family: structures, suggested functions and expression in normal and malignant tissues. Semin Cancer Biol 9:67–81

    Article  PubMed  CAS  Google Scholar 

  14. Herin M, Lemoine C, Weynants P, Vessiere F, Van Pel A, Knuth A, Devos R, Boon T (1987) Production of stable cytolytic T-cell clones directed against autologous human melanoma. Int J Cancer 39:390–396

    Article  PubMed  CAS  Google Scholar 

  15. Hernandez J, Lee PP, Davis MM, Sherman LA (2000) The use of HLA A2.1/P53 peptide tetramers to visualize the impact of self tolerance on the Tcr repertoire. J Immunol 164:596–602

    PubMed  CAS  Google Scholar 

  16. Jessup JM, Petrick AT, Toth CA, Ford R, Meterissian S, O’Hara CJ, Steele GJ, Thomas P (1993) Carcinoembryonic antigen: eanhancement of liver colonisation through retention of human colorectal carcinoma cells. Br J Cancer 67:464–470

    PubMed  CAS  Google Scholar 

  17. Kreuwel HT, Sherman LA (2001) The T-cell repertoire available for recognition of self-antigens. Curr Opin Immunol 13:639–643

    Article  PubMed  CAS  Google Scholar 

  18. Kuroki M, Arakawa F, Matsuo Y, Oikawa S, Misumi Y, Nakazato H, Matsuoka Y (1991) Molecular cloning of nonspecific cross-reacting antigens in human granulocytes. J Biol Chem 266:11810–11817

    PubMed  CAS  Google Scholar 

  19. Machlenkin A, Paz A, Bar Haim E, Goldberger O, Finkel E, Tirosh B, Volovitz I, Vadai E, Lugassy G, Cytron S, Lemonnier F, Tzehoval E, Eisenbach L (2005) Human Ctl epitopes prostatic acid phosphatase-3 and six-transmembrane epithelial antigen of prostate-3 as candidates for prostate cancer immunotherapy. Cancer Res 65:6435–6442

    Article  PubMed  CAS  Google Scholar 

  20. Marshall J (2003) Carcinoembryonic antigen-based vaccines. Semin Oncol 30:30–36

    Article  PubMed  CAS  Google Scholar 

  21. Montes M, Rufer N, Appay V, Reynard S, Pittet MJ, Speiser DE, Guillaume P, Cerottini JC, Romero P, Leyvraz S (2005) Optimum in vitro expansion of human antigen-specific CD8 T cells for adoptive transfer therapy. Clin Exp Immunol 142:292–302

    Article  PubMed  CAS  Google Scholar 

  22. Nagel G, Grunert F, Kuijpers TW, Watt SM, Thompson J, Zimmermann W (1993) Genomic organization, splice variants and expression of Cgm1, a CD66-related member of the carcinoembryonic antigen gene family. Eur J Biochem 214:27–35

    Article  PubMed  CAS  Google Scholar 

  23. Nap M, Mollgard K, Burtin P, Fleuren GJ (1988) Immunohistochemistry of carcino-embryonic antigen in the embryo, fetus and adult. Tumour Biol 9:145–153

    Article  PubMed  CAS  Google Scholar 

  24. Overwijk WW, Tsung A, Irvine KR, Parkhurst MR, Goletz TJ, Tsung K, Carroll MW, Liu C, Moss B, Rosenberg SA, Restifo NP (1998) Gp100/Pmel 17 is a murine tumor rejection antigen: induction of “self”-reactive, tumoricidal T cells using high-affinity, altered peptide ligand. J Exp Med 188:277–286

    Article  PubMed  CAS  Google Scholar 

  25. Parham P, Brodsky FM (1981) Partial purification and some properties of Bb7.2. A cytotoxic monoclonal antibody with specificity for HLA-A2 and a variant of HLA-A28. Hum Immunol 3:277–299

    Article  PubMed  CAS  Google Scholar 

  26. 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 152:163–175

    PubMed  CAS  Google Scholar 

  27. Parkhurst MR, Riley JP, Igarashi T, Li Y, Robbins PF, Rosenberg SA (2004) Immunization of patients with the Htert:540–548 peptide induces peptide-reactive T lymphocytes that do not recognize tumors endogenously expressing telomerase. Clin Cancer Res 10:4688–4698

    Article  PubMed  CAS  Google Scholar 

  28. Princiotta MF, Finzi D, Qian SB, Gibbs J, Schuchmann S, Buttgereit F, Bennink JR, Yewdell JW (2003) Quantitating protein synthesis, degradation, and endogenous antigen processing. Immunity 18:343–354

    Article  PubMed  CAS  Google Scholar 

  29. Ramakrishna V, Ross MM, Petersson M, Gatlin CC, Lyons CE, Miller CL, Myers HE, McDaniel M, Karns LR, Kiessling R, Parmiani G, Flyer DC (2003) Naturally occurring peptides associated with HLA-A2 in ovarian cancer cell lines identified by mass spectrometry are targets of HLA-A2-restricted cytotoxic T cells. Int Immunol 15:751–763

    Article  PubMed  CAS  Google Scholar 

  30. Rammensee H, Bachmann J, Emmerich NP, Bachor OA, Stevanovic S (1999) SYFPEITHI: database for MHC ligands and peptide motifs. Immunogenetics 50:213–219

    Article  PubMed  CAS  Google Scholar 

  31. Rodeberg DA, Nuss RA, Elsawa SF, Celis E (2005) Recognition of six-transmembrane epithelial antigen of the prostate-expressing tumor cells by peptide antigen-induced cytotoxic T lymphocytes. Clin Cancer Res 11:4545–4552

    Article  PubMed  CAS  Google Scholar 

  32. Scardino A, Gross DA, Alves P, Schultze JL, Graff-Dubois S, Faure O, Tourdot S, Chouaib S, Nadler LM, Lemonnier FA, Vonderheide RH, Cardoso AA, Kosmatopoulos K (2002) Her-2/Neu and HTERT cryptic epitopes as novel targets for broad spectrum tumor immunotherapy. J Immunol 168:5900–5906

    PubMed  CAS  Google Scholar 

  33. Schirle MKW, Weber B, Gouttefangeas C, Dumrese T, Becker HD, Stevanovic S, Rammensee HG (2000) Identification of tumor-associated MHC class I ligands by a novel T cell-independent approach. Eur J Immunol 30:2216–2225

    Article  PubMed  CAS  Google Scholar 

  34. Screaton RA, Penn LZ, Stanners CP (1997) Carcinoembryonic antigen, a human tumor marker, cooperates with Myc and Bcl-2 in cellular transformation. J Cell Biol 137:939–952

    Article  PubMed  CAS  Google Scholar 

  35. Shaw AR, Chan JK, Reid S, Seehafer J (1985) Hla-Dr synthesis induction and expression in HLA-Dr-negative carcinoma cell lines of diverse origins by interferon-gamma but not by interferon-beta. J Natl Cancer Inst 74:1261–1268

    PubMed  CAS  Google Scholar 

  36. Skipper JC, Gulden PH, Hendrickson RC, Harthun N, Caldwell JA, Shabanowitz J, Engelhard VH, Hunt DF, Slingluff CLJ (1999) Mass-spectrometric evaluation of HLA-A*0201-associated peptides identifies dominant naturally processed forms of Ctl epitopes from Mart-1 and Gp100. Int J Cancer 82:669–677

    Article  PubMed  CAS  Google Scholar 

  37. Speiser DE, Pittet MJ, Guillaume P, Lubenow N, Hoffman E, Cerottini JC, Romero P (2004) Ex vivo analysis of human antigen-specific CD8(+) T-cell responses: quality assessment of fluorescent HLA-A2 multimer and interferon-gamma elispot assays for patient immune monitoring. J Immunother 27:298–308

    Article  PubMed  CAS  Google Scholar 

  38. Thompson JA, Grunert F, Zimmermann W (1991) Carcinoembryonic antigen gene family: molecular biology and clinical perspectives. J Clin Lab Anal 5:344–366

    Article  PubMed  CAS  Google Scholar 

  39. 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 30:3411–3421

    Article  PubMed  CAS  Google Scholar 

  40. 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 160:1750–1758

    PubMed  CAS  Google Scholar 

  41. Valmori D, Gileadi U, Servis C, Dunbar PR, Cerottini JC, Romero P, Cerundolo V, Levy F (1999) Modulation of proteasomal activity required for the generation of a cytotoxic T lymphocyte-defined peptide derived from the tumor antigen Mage-3. J Exp Med 189:895–906

    Article  PubMed  CAS  Google Scholar 

  42. Van den Eynde B, Hainaut P, Herin M, Knuth A, Lemoine C, Weynants P, van der Bruggen P, Fauchet R, Boon T (1989) Presence on a human melanoma of multiple antigens recognized by autologous CTL. Int J Cancer 44:634–640

    Article  PubMed  Google Scholar 

  43. Van Der Bruggen P, Zhang Y, Chaux P, Stroobant V, Panichelli C, Schultz ES, Chapiro J, Van Den Eynde BJ, Brasseur F, Boon T (2002) Tumor-specific shared antigenic peptides recognized by human T cells. Immunol Rev 188:51–64

    Article  Google Scholar 

  44. Vonderheide RH, Hahn WC, Schultze JL, Nadler LM (1999) The telomerase catalytic subunit is a widely expressed tumor-associated antigen recognized by cytotoxic T lymphocytes. Immunity 10:673–679

    Article  PubMed  CAS  Google Scholar 

  45. Yee C, Savage PA, Lee PP, Davis MM, Greenberg PD (1999) Isolation of high avidity melanoma-reactive Ctl from heterogeneous populations using peptide-Mhc tetramers. J Immunol 162:2227–2234

    PubMed  CAS  Google Scholar 

  46. Yewdell JW (2001) Not such a dismal science: the economics of protein synthesis, folding, degradation and antigen processing. Trends Cell Biol 11:294–297

    Article  PubMed  CAS  Google Scholar 

  47. Zhu MZ, Marshall J, Cole D, Schlom J, Tsang KY (2000) Specific cytolytic T-cell responses to human Cea from patients immunized with recombinant Avipox-Cea vaccine. Clin Cancer Res 6:24–33

    PubMed  CAS  Google Scholar 

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Acknowledgments

This study was supported in part by Swiss National Science Foundation special program NCCR Molecular Oncology. We would like to acknowledge the technical support of Estelle Devevre and Frederic Grosjean and Nicole Montandon.

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Authors and Affiliations

  1. Division of Clinical Onco-Immunology, Ludwig Institute for Cancer Research, Hôpital Orthopédique, HO-05, Rue Pierre-Decker, 4, 1005, Lausanne, Switzerland

    Pedro M. S. Alves, Gabriel Bricard, Daniel E. Speiser & Pedro Romero

  2. Ludwig Institute for Cancer Research, Lausanne Branch, University of Lausanne, Epalinges, Switzerland

    Pedro M. S. Alves, Sebastien Viatte, Theres Fagerberg, Olivier Michielin, Frédéric Lévy & Jean-Charles Cerottini

  3. Molecular Oncology, National Center for Competence in Research (NCCR), Lausanne, Switzerland

    Pedro M. S. Alves & Sebastien Viatte

  4. Institut Universitaire de Pathologie, Centre Hospitalier Universitaire Vaudois, Lausanne, Switzerland

    Hanifa Bouzourene

  5. Department of Surgery, Centre Hospitalier Universitaire Vaudois, Lausanne, Switzerland

    Henri Vuilleumier, Thorsten Kruger & Jean-Claude Givel

  6. Swiss Institute of BioInformatics, Basel, Switzerland

    Theres Fagerberg & Olivier Michielin

  7. Multidisciplinary Oncology Center, Lausanne University Hospital, Lausanne, Switzerland

    Olivier Michielin

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  1. Pedro M. S. Alves
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  2. Sebastien Viatte
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  3. Theres Fagerberg
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  11. Daniel E. Speiser
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  13. Pedro Romero
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Correspondence to Pedro Romero.

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Alves, P.M.S., Viatte, S., Fagerberg, T. et al. Immunogenicity of the carcinoembryonic antigen derived peptide 694 in HLA-A2 healthy donors and colorectal carcinoma patients. Cancer Immunol Immunother 56, 1795–1805 (2007). https://doi.org/10.1007/s00262-007-0323-2

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  • DOI: https://doi.org/10.1007/s00262-007-0323-2

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