Abstract: Objective Aplastic anemia (AA) is considered as an immune-mediated bone marrow failure syndrome with abnormal immune cells and cytokines. In clinic, immunosuppressive therapy (IST) is the first choice for the treatment of severe or very severe aplastic anemia (SAA/VSAA) without suitable hematopoietic stem cell donors. So we aimed to investigate the impact of IST on expression of cytokines in children with acquired SAA. Methods From October 2017 to December 2018, 25 cases of newly diagnosed children with SAA/VSAA was selected in our department, and 37 children for moderate aplastic anemia (MAA). Flow cytometry was used to measure the levels of IFN-γ, TNF-α, IL-2, IL-4, IL-6 and IL-10 in peripheral blood from all, and the changes in the expression of the above cytokines after IST in children with SAA/VSAA in order to evaluate the abnormal immunity in AA indirectly. Results Compared with MAA, children with newly diagnosed acquired SAA/VSAA showed higher levels of IFN-γ (0.84 pg·mL-1 vs 0.37 pg·mL-1, P=0.009 7) and IL-6 (47.26 pg·mL-1 vs 13.54 pg·mL-1, P=0.046 8). After IST, total 25 cases of complete remission and partial remission were obtained, and the effective rate of IST was 100%. And six months after IST, the expressions of IFN-γ (0.02 pg·mL-1 vs 0.56 pg·mL-1), TNF-α (0.88 pg·mL-1 vs 11.51 pg·mL-1) and IL-6 (9.60 pg·mL-1 vs 71.44 pg·mL-1) were decreased significantly in children with SAA/VSAA (P<0.05). One year after IST, the concentrations of above cytokines were decreased to some extent. Conclusion Children with newly diagnosed acquired SAA/VSAA showed abnormal levels of cytokines in serum. However, IST could significantly improve the expression of these negative hematopoietic regulatory factors.