The impact of anti-HLA antibodies and crossmatch (CM) on liver transplantation (LT) outcome is still controversial. In this retrospective study we analyzed LT outcome according to pretransplant pre-formed anti-HLA antibodies and CM status. Serum anti-HLA antibodies were screened by ELISA assay, utilizing One Lambda antigen tray-mixed (LAT-M). CMs were performed by the complement dependent cytotoxicity test using Dithiotreitol treated sera. Anti-HLA antibodies were studied in 80 recipients; 56/80 had positive LAT-M tests (PLAT-M), whereas the remaining 24 recipients tested negative for both classes I and II (NLAT-M). Rejection episodes were more frequent in PLAT-M compared with NLAT-M group in post-LT intervals of <1 week (p = 0.05), 1 week-3 months (p = 0.035), and 3-12 months (p = 0.076). Graft and patient survival rates were better, albeit not significantly, in the NLAT-M compared with PLAT-M recipients. CM status was investigated in 62/80 recipients, 18/62 recipients had positive CM (PCM), and 44 had negative CM (NCM). Five of 18 PCM recipients (28%) experienced early graft loss compared with 1/44 (2%) with NCM (p = 0.006). Rejection episodes were more frequent within first 3 months post-LT in PCM recipients compared with NCM (p = 0.015). One-year graft survival rate was better in NCM, compared with PCM recipients (graft loss of 2/44 vs 5/18). NCM PLAT-M had a higher incidence of rejection episodes compared with the NCM NLAT-M group (p = 0.031). The presence of anti-HLA antibodies suggests a deleterious effect on LT outcome, and was associated with an increased incidence of early graft loss and rejection episodes.