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Increased PRAME antigen-specific killing of malignant cell lines by low avidity CTL clones, following treatment with 5-Aza-2′-Deoxycytidine

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Abstract

The cancer testis antigen Preferentially Expressed Antigen of Melanoma (PRAME) is overexpressed in many solid tumours and haematological malignancies whilst showing minimal expression in normal tissues and is therefore a promising target for immunotherapy. HLA-A0201-restricted peptide epitopes from PRAME have previously been identified as potential immunogens to drive antigen-specific autologous CTL responses, capable of lysing PRAME expressing tumour cells. CTL lines, from 13 normal donors and 10 melanoma patients, all of whom were HLA-A0201 positive, were generated against the PRAME peptide epitope PRA100−108. Specific killing activity against PRA100−108 peptide-pulsed targets was weak compared with CTL lines directed against known immunodominant peptides. Moreover, limiting dilution cloning from selected PRAME-specific CTL lines resulted in the generation of a clone of only low to intermediate avidity. Addition of the demethylating agent 5-aza-2′-Deoxycytidine (DAC) increased PRAME expression in 7 out of 11 malignant cell lines including several B lineage leukaemia lines and also increased class I expression. Pre-treatment of target cells was associated with increased sensitivity to antigen-specific killing by the low avidity CTL. When CTL, as well as of the target cells, were treated, the antigen-specific killing was further augmented. Interestingly, one HLA-A0201-negative DAC-treated line (RAJI) showed increased sensitivity to killing by clones despite a failure of expression of PRAME or HLA-A0201. Together these data point to a general increased augmentation of cancer immunogenocity by DAC involving both antigen-specific and non-specific mechanisms.

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Acknowledgments

Supported by research grants from SPARKS, the Leukaemia Research Fund (UK), Children With Leukaemia and RICC.

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Authors and Affiliations

  1. Unit of Molecular Haematology and Cancer Biology, University College London Institute of Child Health, 30 Guilford Street, London, WC1N 1EH, UK

    Mengyong Yan, Nourredine Himoudi, B. Piku Basu, Rebecca Wallace, Edmund Poon, Fyeza Hasan, Owen Williams & John Anderson

  2. Department of Haematology, Great Ormond Street Hospital, London, UK

    Stuart Adams

  3. Department of Immunology, University College London Royal Free Campus, London, UK

    Shao-An Xue

  4. Department of Medical Oncology, St George’s Hospital Medical School, London, UK

    Natalie Wilson & Angus Dalgleish

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  1. Mengyong Yan
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Correspondence to Owen Williams or John Anderson.

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Yan, M., Himoudi, N., Basu, B.P. et al. Increased PRAME antigen-specific killing of malignant cell lines by low avidity CTL clones, following treatment with 5-Aza-2′-Deoxycytidine. Cancer Immunol Immunother 60, 1243–1255 (2011). https://doi.org/10.1007/s00262-011-1024-4

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