OBJECTIVE This study aimed to evaluate the prevalence and significance of serum autoantibodies in alcoholic liver disease (ALD) patients. METHODS In total, 90 male patients diagnosed with ALD or ALD overlapping with chronic hepatitis B (CHB) were included and their medical records were retrospectively reviewed. Clinical data and laboratory findings were collected. Anti-nuclear antibody (ANA), anti-mitochondrial antibody (AMA) and anti-smooth muscle antibody (SMA) were detected by indirect immunofluorescent assay. RESULTS Autoantibodies were found in 69.6% (48/69) of patients with ALD and 66.7% (14/21) of those with ALD overlapping with CHB. The prevalence of total ANA in ALD patients was 63.8% (44/69). High titers of autoantibodies and multi-autoantibodies were found to be more often associated with cirrhosis than non-cirrhosis. There was a significant difference in the levels of globulin, white blood cell and platelet count (P < 0.05) between patients with positive and negative autoantibodies. However, the values of the other parameters were similar in the two groups. Patients with ALD more frequently had positive autoantibodies than those with CHB alone (69.6% vs. 37.5%, P < 0.01). And 10.4% of ALD patients with positive autoantibodies had systemic autoimmune or vascular diseases, which was not found in the CHB patients. CONCLUSIONS Autoantibodies are frequently present in patients with ALD and correlate to advanced liver disease. A high prevalence of autoantibodies in ALD may indicate that alcoholic-associated immune disturbance occurs during the development of the disease.