databases), combined with the novel adjuvant CAF09b and used for the immunisation of HHDII/DR1 and HHDII/DP4 mice. Following splenocyte extraction, peptides-derived vaccines were tested for immunogenicity using IFNg ELISpot assays and killing of target cells was assessed using the same assay, co-culturing isolated CD3 + T cells with antigen-expressing cancer cells. Results Antigen expression of TNBC cell lines as revealed by qPCR suggested that treatment with 5mM DAC induced optimal upregulation, with 10mM usually leading to a plateau. These observations were confirmed by western blotting. Splenocytes derived from CAF09b-adjuvanted NY-ESO-1/HAGE/ WT1 vaccine-immunised mice showed strong IFNgresponses, and the isolated CD3 + T cells from these splenocytes could specifically recognise TNBC cells expressing all three antigens in a HLA-A2 restricted manner. Conclusions We propose the use of mutated NY-ESO-1/HAGE/ WT1-derived peptides in combination with CAF09b for the treatment of GBM, PCa and TNBC at a stage where no tumour cells can be detected to prevent or delay relapse in these malignancies.
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