Aim: Hashimoto's thyroiditis is part of the spectrum of autoimmune thyroid diseases characterized by the destruction of thyroid cells by antibody-mediated immune processes. Although there are some studies about the association of macroprolactinaemia and interleukin-21 (IL-21) levels with other autoimmune diseases, few studies have been found in association with the Hashimoto's thyroiditis in the literature. In this study, we aimed to see if there is any association between IL-21 and macroprolactin levels with antithyroid antibodies. Subjects and Method: The study included 70 patients with Hashimoto's thyroiditis as the study group and 48 healthy volunteers as the control group. Interleukin-21, prolactin, free thyroxine, thyroid-stimulating hormone (TSH), antibodies against thyroglobulin (TG) and anti-thyroid peroxidase (anti-TPO) were determined in all participants. The presence of macroprolactinaemia was manifested by a polyethylene glycol precipitation method. Results: There was no significant difference between the study and the control groups for free triiodothyronine (T3), free thyroxine (T4), prolactin and macroprolactin levels. We could not find any relationship between prolactin and macroprolactin levels with anti-TPO and anti-thyroglobulin (anti-TG) levels. Although IL-21 levels of the control group were significantly higher than the study group, there was also no correlation between IL-21 and anti-TPO and anti-TG levels. Conclusion: Our results showed that neither macroprolactin nor prolactin levels have any relationship with anti-thyroid antibodies. Contrary to other studies, IL-21 levels of the control group were higher than the study group. We think that should be done with larger groups to better understand if there was any relationship of IL-21 and macroprolactin levels with Hashimoto's thyroiditis.