Identification of novel myeloma-specific XBP1 peptides able to generate cytotoxic T lymphocytes: A potential therapeutic application in multiple myeloma

The purpose of these studies was to identify HLA-A2+ immunogenic peptides derived from XBP1 antigens to induce a multiple myeloma (MM)-specific immune response. Six native peptides from non-spliced XBP1 antigen and three native peptides from spliced XBP1 antigen were selected and evaluated for their HLA-A2 specificity. Among them,\( XBP1_{184–192}\), XBP1 \(SP_{196–204}\) and XBP1 \(SP_{367–375}\) peptides showed the highest level of binding affinity, but not stability to HLA-A2 molecules. Novel heteroclitic XBP1 peptides, YISPWILAV or YLFPQLISV, demonstrated a significant improvement in HLA-A2 stability from their native \(XBP1_{184–192}\) or XBP1 \(SP_{367–375}\) peptide, respectively. Cytotoxic T lymphocytes generated by repeated stimulation of CD3+ T cells with each HLA-A2-specific heteroclitic peptide showed an increased percentage of CD8+ (cytotoxic) and CD69+/CD45RO+ (activated memory) T cells and a lower percentage of CD4+ (helper) and CD45RA+/CCR7+ (naïve) T cells, which were distinct from the control T cells. Functionally, the CTLs demonstrated MM-specific and HLA-A2-restricted proliferation, IFN-γ secretion and cytotoxic acivity in response to MM cell lines and importantly, cytotoxicty against primary MM cells. These data demonstrate the distinct immunogenic characteristics of unique heteroclitic XBP1 peptides which induce MM-specific CTLs and highlights their potential application for immunotherapy to treat the patients with MM or its pre-malignant condition