Resistance or relapse upon chemotherapy are major determinants of treatment failure in acute myeloid leukemia (AML). Therapy-induced senescence (TIS) is a potential outcome of chemotherapy, but its immunological consequences in AML remain unclear. We show that ex-vivo chemotherapy induces senescence in a subset of therapy-naïve AML samples. TIS is marked by elevated interferon signaling, upregulation of HLA class I/II molecules, and increased presentation of leukemia and senescence-associated peptides, conferring AML blasts antigen-presenting cell-like features. Patient-derived leukemic blasts purified from AML samples were either treated with cytarabine (AraC) or left untreated. Further, samples that exhibited a significant increase in fluorescent SA-β-Gal upon treatment were classified as Senescence High (Sen High), whereas others that showed minimal change were classified as Senescence Low (Sen Low). We employed our in-house microfluidic platform, PeptiCHIP for immunoaffinity purification followed by Ligandomics LC/MS analysis, to identify HLA class I-bound peptides in Patient-derived leukemic blasts, seeking to compare the immunopeptidome repertoire of Sen Low and Sen High samples before and after cytarabine treatment. Sen High exhibited a greater number of eluted peptides compared to Sen Low samples already at baseline. This number was further increased following chemotherapy in all Sen High samples analyzed. Conversely, the number of peptides retrieved in Sen Low samples remained low regardless of chemotherapy treatment.