This study aims to investigate the histopathological expression of Interferon-gamma (IFN-gamma) and CD-56 in pediatric patients who underwent surgery for adenoid hypertrophy, and to evaluate their association with coexisting otitis media with effusion (OME) and tonsillar hypertrophy. A total of 100 children who underwent adenoidectomy, adenotonsillectomy, adenoidectomy with ventilation tube insertion, or adenotonsillectomy with ventilation tube insertion were included and divided into four equal groups. Surgical specimens were stained with hematoxylin-eosin, CD-56, and IFN-gamma. Clinical data including age, sex, comorbidities, adenoid volume, tonsil grade, and presence of OME were analyzed. Immunohistochemical staining rates were compared among the groups. The rate of IFN-gamma staining was significantly higher in the group that underwent adenotonsillectomy with ventilation tube insertion compared to other groups (p < 0.05). A statistically significant increase in IFN-gamma staining was also observed in patients with OME (p < 0.05). CD-56 staining did not show significant differences among the groups (p > 0.05). The elevated IFN-gamma expression in patients with OME and tonsillar hypertrophy may indicate an active inflammatory response. These findings suggest that IFN-gamma may play a role in the pathogenesis of OME in children with adenoid hypertrophy and could be considered as a potential therapeutic target.