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The role of anti–human leukocyte antigen (HLA) donor-specific antibodies (DSA) detected by Luminex in the development of rejection is not fully understood.A study including 369 patients who received transplants from deceased donors with a negative complement-dependent cytotoxicity crossmatch (XM) was performed. From the total of patients, 151 underwent a renal biopsy because of renal dysfunction, whereas the 218 remaining showed a stable renal function, and no rejection was assumed. Diagnosis and type of rejection was based in biopsy data.Patients with a positive virtual XMs showed more rejection episodes of any types when comparing with patients with negative virtual XMs (P<0.0001). Nevertheless, there were no significant differences between patients without anti-HLA antibodies and patients with anti-HLA no DSA. Allograft impairment was caused by a rejection episode in 84% (32/38) of patients with anti–HLA-DSA but only in 30% (34/113) of patients without anti–HLA-DSA. Regarding the type of rejection detected in the biopsy, all the patients with de novo (after transplantation) anti–HLA-DSA were diagnosed as antibody-mediated rejection (AMR) or AMR+T-cell–mediated rejection, whereas most of the patients without anti–HLA-DSA (68%) were diagnosed with T-cell–mediated rejection, and patients with preexistent anti–HLA-DSA showed a more homogeneous distribution of the different types of rejection.According to our results, patients with preformed or de novo anti–HLA-DSA showed the highest likelihood to suffer rejection episodes. Transplantation with preformed anti–HLA-DSA should be avoided, and an early detection of de novo HLA antibodies is important to treat patients before damage occurs in the graft.