Introduction: Although autoimmune thyroid disease is less common in males, it is unclear whether estrogen contributes to the difference in susceptibility among males. Objective: To examine whether circulating estradiol (E 2 ) is related to thyroid autoimmunity in males. Patients and methods: One-thousand two-hundred and sixty-three males aged 15-94 years were studied. Serum levels of E 2 , TSH receptor antibody (TRAb), thyroid peroxidase antibody (TPOAb), thyroglobulin antibody (TgAb), free thyroxine (FT 4 ), and TSH were measured by ELISA. Results: Circulating E 2 varied widely in males, ranging 18.4-403.7 pmol/l with a mean value of 136.2G51.7 pmol/l. E 2 increased with age (rZ0.18, P!0.001). No relationship between E 2 and BMI was found. When comparing the difference in E 2 according to the test results of TRAb, TPOAb, and TgAb, it was found that E 2 was significantly higher in subjects with positive TRAb (TRAb positive, E 2 Z170.3G59.8 pmol/l; TRAb negative, E 2 Z134.0G50.6 pmol/l; P!0.001). No difference in E 2 was demonstrated according to the results of TPOAb or TgAb. Logistic regression analysis showed that E 2 was a determinant of positive TRAb, independent of age and BMI. There was no relationship between serum E 2 and TSH or FT 4 . However, E 2 was negatively related to TSH (rZK0.45, P!0.01) in subjects whose TSH levels fell below the reference range (0.3-4.2 mIU/l).