Graves' disease (GD) represents one of the most prevalent autoimmune disorders affecting the thyroid gland. Our investigation aimed to evaluate serum levels and the DNA methylation in the promoter regions of INF-γ in patients with GD compared to healthy controls. Methods: Blood samples were collected to measure serum levels of TSH, TPO antibodies, and cytokines (INF-γ), followed by DNA extraction and bisulfite conversion for methylation analysis using Methylation-Specific PCR (MSP). Results: Non-significant differences were noted between groups regarding age distribution or gender. Serum analysis revealed significantly lower mean TSH levels in GD patients, Additionally, there is statistically significant increase in INF-γ among GD patients compared to controls. Analysis of DNA methylation patterns showed hypomethylated INF-γ promoter regions with statistically non-significant difference between the two groups. Conclusion: INF-γ is elevated significantly in GD patients and promoter regions were hypomethylated, although the difference in methylation did not reach statistical significance. This finding indicates that methylation in INF-γ promoter may has a critical role in the gene regulation.