Amino-acid sequences of TCRα chain CDR3 regions of Ep63K-specific CD4⁺ T cell hybridomas obtained from TCR^mini-Foxp3^GFP mice and generation of B16 tumors expressing tumor associated neo-antigen NP-Ep63K.

<p>(A) Amino-acid sequences of TCRα chain CDR3 regions of Ep63K-specific CD4⁺ T cell hybridomas obtained from TCR^mini-Foxp3^GFP mice. (B) Production of the nucleoprotein-Ep63K-GFP expression construct. Sections encoding nucleoprotein are shown as dotted rectangles and GFP is shown as vertical lines. Parts of the nucleoprotein encoding peptides binding A^b are shown as clear rectangles and Ep63K peptide is shown as horizontal lines. Numbers show the range of amino acids constituting peptides binding to A^b. (C) Flow cytometry analysis of the B16 melanoma cells expressing nucleoprotein-Ep63K-GFP. B16 melanoma cells were transduced with the LZRS-pBMN-Z retroviral vector expressing nucleoprotein-Ep63K-GFP fusion protein (lower histogram) or a control vector (upper histogram). (D) Recombinant nucleoprotein-Ep63K is processed by bone marrow derived dendritic cells and Ep63K peptide is recognized by specific CD4⁺ hybridoma 123.3. IL-2 (pg/ml) production in the supernatant of dendritic cells cultured without and with 0.9 or 1.8 µg/ml of the recombinant protein.