Fig. 4.

Conserved IFN-γ immunity in ZNFX1-deficient patients. PBMCs from controls (green dots), age-matched controls (blue dots), and patients with AR ZNFX1 deficiency (red dots) were left unstimulated or were stimulated with IL-12, or IL-23, with or without bacillus Calmette–Guérin activation. (A) IFN-γ levels in the supernatant of PBMCs with and without stimulation with IL-12, IL-23, bacillus Calmette–Guérin, bacillus Calmette–Guérin+IL-12, and bacillus Calmette–Guérin+IL-23, as assessed by intracellular flow cytometry. Proportion of IFN-γ−producing lymphocytes of various subsets involved in innate (NK, iNKT, and MAIT cells), adaptive (CD4⁺ and CD8⁺ T cells), and both adaptive and innate (γδ T cells including Vδ1⁺, Vδ2⁺, and Vδ1⁻Vδ2⁻ subsets) immunity after stimulation with (B) bacillus Calmette–Guérin, (C) IL-12, (D) bacillus Calmette–Guérin+ IL-12, (E) IL-23, or (F) bacillus Calmette–Guérin+IL-23. (G) Secretion of IL-12p40 by whole blood from daily controls (n = 69), travel controls (n = 14), P1, with and without stimulation with bacillus Calmette–Guérin alone, or bacillus Calmette–Guérin and IFN-γ. ELISA was used to determine the levels of this cytokine. (H) Secretion of IL-12p40 by EBV-B cells from controls, patients with AR complete IL-12p40 deficiency (IL-12p40^−/−), or P1, after stimulation with 10⁻⁷ M PDBu for 24 h. ELISA was used to determine the levels of these cytokines. (I) Phosphorylation of STAT1 after 20 min of stimulation with IFN-γ (10⁴ IU/mL) or IFN-α (10⁴ IU/mL) in EBV-B cells from a healthy control (C+), P1, and a patient with AR complete STAT1 deficiency (STAT1^−/−), as determined by flow cytometry. (J) Induction of HLA-DR expression in SV40 fibroblasts from controls (C+), P1, P2, and a patient with AR complete IFN-γR1 deficiency (IFN-γR1^−/−) after 48 h of stimulation with IFN-γ (10⁴ IU/mL), as determined by flow cytometry. The results of I and J are representative of two independent experiments.