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Low TCR avidity and lack of tumor cell recognition in CD8+ T cells primed with the CEA-analogue CAP1-6D peptide

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Abstract

The use of “altered peptide ligands” (APL), epitopes designed for exerting increased immunogenicity as compared with native determinants, represents nowadays one of the most utilized strategies for overcoming immune tolerance to self-antigens and boosting anti-tumor T cell-mediated immune responses. However, the actual ability of APL-primed T cells to cross-recognize natural epitopes expressed by tumor cells remains a crucial concern. In the present study, we show that CAP1-6D, a superagonist analogue of a carcinoembriyonic antigen (CEA)-derived HLA-A*0201-restricted epitope widely used in clinical setting, reproducibly promotes the generation of low-affinity CD8+ T cells lacking the ability to recognized CEA-expressing colorectal carcinoma (CRC) cells. Short-term T cell cultures, obtained by priming peripheral blood mononuclear cells from HLA-A*0201+ healthy donors or CRC patients with CAP1-6D, were indeed found to heterogeneously cross-react with saturating concentrations of the native peptide CAP1, but to fail constantly lysing or recognizing through IFN- γ release CEA+CRC cells. Characterization of anti-CAP1-6D T cell avidity, gained through peptide titration, CD8-dependency assay, and staining with mutated tetramers (D227K/T228A), revealed that anti-CAP1-6D T cells exerted a differential interaction with the two CEA epitopes, i.e., displaying high affinity/CD8-independency toward the APL and low affinity/CD8-dependency toward the native CAP1 peptide. Our data demonstrate that the efficient detection of self-antigen expressed by tumors could be a feature of high avidity CD8-independent T cells, and underline the need for extensive analysis of tumor cross-recognition prior to any clinical usage of APL as anti-cancer vaccines.

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Abbreviations

APL:

Altered peptide ligands

CEA:

Carcinoembriyonic antigen

CRC:

Colo-rectal carcinoma

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Acknowledgments

We wish to thank Dr Jorgelina Coppa and Dr Vincenzo Mazzaferro (Unit of Liver Transplant, Istituto Nazionale Tumori) for providing patients’ blood samples, and Dr. Italia Bongarzone (Proteomics Laboratory, Istituto Nazionale Tumori) for performing mass-spectrometry analysis of peptide preparations. We are also grateful to Gloria Sovena and Agata Cova for the excellent technical support. Grant support: Grant from the European Community, VI Program, Cancer Immunotherapy Project, n°518234; Italian Ministry of Industry, University and Research (MIUR), grant FIRB # RBNE017B4C (Rome, Italy); Italian Association for Cancer Research (AIRC, Milan, Italy).

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  1. Giorgio Parmiani

    Present address: Unit of Immunobiotherapy of Solid Tumors, San Raffaele Scientific Institute, Via Olgettina 58, 20132, Milan, Italy

Authors and Affiliations

  1. Unit of Immunotherapy of Human Tumors, IRCCS Istituto Nazionale Tumori, Via Venezian 1, 20133, Milan, Italy

    Manuela Iero, Paola Squarcina, Matteo Carrabba, Giorgio Parmiani & Licia Rivoltini

  2. Division of Clinical Onco-Immunology, Ludwig Institute for Cancer Research, Lausanne Branch, Lausanne, Switzerland

    Pedro Romero & Philippe Guillaume

  3. Molecular and Cellular Biology Department, IRBM P. Angeletti, Pomezia, Italy

    Elisa Scarselli & Raffaele Cerino

  4. Laboratoire de Cytogénétique et de Biologie Cellulaire, Hôpital Pontchaillou, Rennes, France

    Olivier Toutirais

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  1. Manuela Iero
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  2. Paola Squarcina
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Correspondence to Licia Rivoltini.

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Iero, M., Squarcina, P., Romero, P. et al. Low TCR avidity and lack of tumor cell recognition in CD8+ T cells primed with the CEA-analogue CAP1-6D peptide. Cancer Immunol Immunother 56, 1979–1991 (2007). https://doi.org/10.1007/s00262-007-0342-z

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