TNF-α expression by BM macrophages from AA patients and impact of TNF-α on human T-cell IFN-γ expression. (A) Comparison of the frequencies of BM macrophages (CD16⁺CD68⁺ in viable CD3⁻ population) in AA patients (n = 8) and healthy controls (HC; n = 7). (B) CD14 and CD16 expression in CD16⁺CD68⁺ population. (C) Frequencies of TNF-α–producing cells in CD16⁺CD68⁺, CD4⁺, and CD8⁺ populations in AA patients detected by intracellular staining. Representative dot plots are shown. (D) Peripheral blood mononuclear cells (2 × 10⁶/mL) from AA patients or healthy controls were stimulated with PMA (5 ng/mL) plus ionomycin (1 μM) in the presence or absence of human recombinant TNF-α (100 ng/mL) overnight. Intracellular IFN-γ levels in CD4⁺ T cells and CD8⁺ T cells from healthy controls (n = 7) and AA patients (n = 8) were examined by flow cytometry. *P < .05; **P < .01.