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The diagnosis of humoral rejection in lung transplant recipients is sometimes elusive, involving histopathologic (capillaritis), immunopathologic (C4d staining), and serologic (donor-specific anti-HLA antibodies, DSA) parameters. These criteria are not always present in cases and, moreover, the involvement of non-HLA antibodies in lung-allograft humoral rejection remains largely unclear. The aim of the study was to assess the involvement of HLA and non-HLA antibodies in humoral rejection diagnosis after lung transplantation.A total of 60 consecutive lung transplant recipients were followed in our Institution from 2015. Protocol biopsy at day 21 post transplantation and screening for serum HLA and non-HLA antibodies by Luminex, LABScreen Autoantibody (One Lambda), before lung transplant and at the moment of the biopsy, were performed.A total of 5 lung transplant recipients (8.33%) were C4d+ on biopsy but only one of them developed de novo DSA without evidence of non-HLA antibodies. The remaining 4 patients C4d+, were serum positive for non-HLA antibodies (2 against LG3 and other 2 against myosin) prior lung transplantation (6.67%) but negative at the moment of the biopsy. The incidence observed of preformed non-HLA antibodies in patients awaiting for lung transplantation was significant and potentially could drive C4d deposition on lung tissue with subsequent humoral rejection.The screening of non-HLA and HLA antibodies could help to identify patient at risk of lung-allograft rejection.