Specific recognition and inhibition of Ewing tumour growth by antigen-specific allo-restricted cytotoxic T cells

Background:

The development of a successful immunotherapy is hampered by an ineffective T-cell repertoire against tumour antigens and the inability of the patient''s immune system to overcome tolerance-inducing mechanisms. Here, we test the specific recognition and lytical potential of allo-restricted CD8⁺ T cells against Ewing tumour (ET) associated antigens Enhancer of Zeste, Drosophila Homolog 2 (EZH2), and Chondromodulin-I (CHM1) identified through previous microarray analysis.

Methods:

Following repetitive CHM1³¹⁹ (VIMPCSWWV) and EZH2⁶⁶⁶ (YMCSFLFNL) peptide-driven stimulations with HLA-A^*0201⁺ dendritic cells (DC), allo-restricted HLA-A^*0201⁻ CD8⁺ T cells were stained with HLA-A^*0201/peptide multimers, sorted and expanded by limiting dilution.

Results:

Expanded T cells specifically recognised peptide-pulsed target cells or antigen-transfected cells in the context of HLA-A^*0201 and killed HLA-A^*0201⁺ ET lines expressing the antigen while HLA-A^*0201^– ET lines were not affected. Furthermore, adoptively transferred T cells caused significant ET growth delay in Rag2^−/−γ_C^−/− mice. Within this context, we identified the CHM1³¹⁹ peptide as a new candidate target antigen for ET immunotherapy.

Conclusion:

These results clearly identify the ET-derived antigens, EZH2⁶⁶⁶ and CHM1³¹⁹, as suitable targets for protective allo-restricted human CD8⁺ T-cell responses against non-immunogenic ET and may benefit new therapeutic strategies in ET patients treated with allogeneic stem cell transplantation.