Objective: To evaluate the expression levels of interferon-γ (IFN-γ), interleukin-2 (IL-2) and interleukin-6 (IL-6) in elderly patients with pulmonary tuberculosis, and analyze the diagnostic value of individual and combined detection of these three indicators. Methods: A retrospective study was conducted. Clinical data were collected from 475 suspected elderly pulmonary tuberculosis patients aged ≥60 years admitted to Lanzhou Pulmonary Hospital from 2023 to 2025. The data included baseline data, imaging findings, etiological results, biochemical tests as well as serum IFN-γ, IL-2 and IL-6 results. Subjects were divided into pulmonary tuberculosis group and non-tuberculosis group based on final clinical diagnosis. Difference in expression levels of the three cytokines between two groups were compared, and the clinical auxiliary value of detecting serum IFN-γ, IL-2, and IL-6 individually or in combination for diagnosing elderly pulmonary tuberculosis was evaluated. Results: Among 475 elderly subjects, 234 were clinically diagnosed with pulmonary tuberculosis and 241 were non-tuberculosis patients. Serum expression levels of IFN-γ (23.14 (2.73, 95.52) pg/ml), IL-2 (17.64 (1.57, 115.05) pg/ml) and IL-6 (19.14 (6.85, 60.86) pg/ml) in tuberculosis group were significantly higher than those in non-tuberculosis group (1.29 (0.00, 5.31), 1.97 (0.00, 7.46), 7.00 (2.18, 16.49) pg/ml), with statistically significant differences (Z=120.866, 80.050, 31.786, Ps<0.001). The sensitivities of single testing for IFN-γ, IL-2 and IL-6 were 59.83% (140/234), 55.56% (130/234) and 56.84% (133/234), specificities were 83.40% (201/241), 84.65% (204/241) and 74.69% (180/241), accuracies were 71.79% (341/475), 70.32% (334/475) and 65.89% (313/475), and corresponding AUC values were 0.74, 0.71 and 0.69 respectively. For combined testing, the sensitivities of IFN-γ+IL-2, IFN-γ+IL-6, IL-2+IL-6 and three-marker panel were 58.97% (138/234), 73.93% (173/234), 67.52% (158/234) and 77.78% (182/234). Their specificities were 90.87% (219/241), 73.86% (178/241), 80.91% (195/241) and 83.40% (201/241), and their accuracies were 75.16% (357/475), 73.89% (351/475), 74.32% (353/475) and 80.63% (383/475). AUC values were 0.76, 0.80, 0.81 and 0.83, respectively. Conclusion: The serum expression levels of IFN-γ, IL-2, and IL-6 in elderly patients with pulmonary tuberculosis were significantly higher than those in non-tuberculosis patients. The combined testing of three markers demonstrated markedly superior diagnostic performance compared to single testing, offering significant clinical value for the auxiliary diagnosis of elderly pulmonary tuberculosis cases.