The present study was undertaken to develop a relatively quantitative enzyme-linked immunosorbent assay (ELISA) in-house using human leukocyte antigen class II-restricted epitopes in order to test circulating autoantibodies to human forkhead/winged helix transcription factor (FOXP3) as a biomarker for esophageal cancer. A total of 97 patients with esophageal squamous cell carcinoma (ESCC) and 227 healthy subjects were recruited for this study, and their plasma samples were collected for antibody analysis with the ELISA approach. Student's t test showed that the anti-FOXP3 IgG antibody levels were significantly higher in the patient group than the control group (t=6.23, P<0.0001). Based on a cutoff value determined by the mean+3SD of control IgG levels, the positive rate was 5.15 % in patients with ESCC as compared to 0.88 % in control subjects (X (2) =6.53, P=0.019, OR=5.85, 95 % CI 1.12-30.67), in which patients at stage I had the highest positivity (11.54 %, X (2) =12.15, P=0.0005, OR=13.10, 95 % CI 2.09-82.04). The sensitivity against >95 % specificity was 22.7 % for the IgG assay with an inter-assay deviation of 13.35 %. This work suggests that circulating IgG autoantibody to FOXP3 may be a potential biomarker for early diagnosis of esophageal cancer.