Identification of an immunogenic DKK1 long peptide for immunotherapy of human multiple myeloma

Dickkopf-1 (DKK1), broadly expressed by tumor cells from human multiple myeloma (MM) and other cancers but absent from most normal tissues, may be an ideal target for immunotherapy. Our previous studies have shown that DKK1 (peptide)-specific cytotoxic T lymphocytes can effectively lyse primary MM cells in vitro. To develop DKK1-based vaccines that can be easily and inexpensively made and used by all patients, we identified a DKK1 long peptide (LP), DKK1(3-76)-LP, that contains 74 amino acids and epitopes that can potentially bind to all major MHC class I and II molecules. Using HLA-A*0201- and HLA-DR*4-transgenic mouse models, we found that DKK1-specific CD4(+) and CD8(+) T-cell responses, detected by DKK1 short peptide (P20 and P66v)-HLA-A*0201 tetramer staining and cytotoxic assay for CD8(+) T cells or by carboxyfluorescein diacetate succinimidyl ester (CSFE) dilution and IFN-g secretion for CD4(+) T cells, respectively, can be induced in vivo by immunizing mice with the DKK1(3-76)-LP. In addition, DKK1(3-76)-LP also induced anti-DKK1 humoral immunity in the transgenic mice and the DKK1 antibodies were functional. Finally, DKK1(3-76)-LP stimulated human blood T cells ex vivo to generate DKK1-specific CD4(+) and CD8(+) T-cell responses from 8 out of 10 MM patients with different MHC backgrounds. The generated DKK1-specific CD8(+) cells efficiently lysed autologous MM cells from these patients. Thus, these results confirm the immunogenicity of the DKK1(3-76)-LP in eliciting DKK1-specific CD4(+) and CD8(+) T-cell responses in vitro and in vivo, and suggest that the DKK1(3-76)-LP can be used for immunotherapy of MM and other cancers.