Platelet transfusion is important in the prevention and treatment of bleeding in patients with acute myeloid leukemia (AML) after receiving intensive chemotherapy. However, platelet transfusion refractoriness (PTR) is an intractable clinical issue occurred in these patients. And its clinical and immunological features remain largely unknown. The potential causes and clinical features of PTR were retrospectively analyzed in 560 patients who were diagnosed as de novo AML in Tongji Hospital from June 2012 through June 2018. A high-throughput antibody screening for the detection of human leukocyte antigen (HLA) and its serotypes was performed in 133 newly diagnosed AML patients. PTR occurred in 11.8% of the de novo AML patients. The median age for patients with PTR was 46 years (range, 15-70). It frequently manifested in female patients and in patients with splenomegaly, M4 subtype, c-Kit gene mutation, and rearrangements of RUNX1-RUNX1T1 or CBFB-MYH11, commonly referred to as core binding factor AML (CBF-AML). Notably, CBF-AML was independently associated with the occurrence of PTR. PTR predominantly developed in patients who had CBF-AML (P < .001) and in patients who further had better minimal residual disease (MRD) reduction (>= 3-log) before the second consolidation chemotherapy (P = .007). HLA-I antibodies were detected in the serum of 9.0% of AML patients and markedly enriched in patients with PTR (P < .001) and in patients with CBF-AML (P = .018). HLA-B was the most frequently identified serum epitope in PTR patients. Patients with CBF-AML had higher tendency to develop HLA-I antibodies and PTR, which depicted novel features of PTR in AML and might provide insights into its efficient managements.