SummaryThe interpretation of the role of HLA‐DPB1 in unrelated haematopoietic stem cell transplantation (HSCT) is subject to discussion. We have investigated the role of HLA‐DPB1 allele matching in HSCT outcomes in 161 recipients who were HLA‐A, ‐B, ‐C, ‐DRB1 and ‐DQB1‐matched with their unrelated donors at the allelic level (10/10). In addition, we analysed the association of polymorphic amino acid mismatches of DPB1 molecule with HSCT end‐points, and a previously published permissiveness concept. HLA‐DPB1 allele mismatches were significantly associated with an increased incidence of acute graft‐versus‐host disease (aGvHD) and worse overall survival (OS). The mismatch at amino acid position 69 significantly increased the risk for transplant‐related mortality (TRM). Risk factors for aGvHD also included mismatches at positions 8, 9, 35, 76 and 84. This is to our knowledge, the first report of an in vivo effect of single amino acid mismatches on HSCT outcomes. In this study, grouping of allelic mismatches into permissive and non‐permissive categories and their association with transplantation end‐points was relevant for TRM but not for other clinical end‐points.