Fig. 4.
IP-10 induction after the stimulation of whole blood from patients with impaired type I IFN-dependent immunity. (A) IP-10 induction after the stimulation of whole blood from five APS-1 patients and nine healthy donors with 10 ng/mL glycosylated IFN-α2, IFN-β, or IFN-ω for 16 h. IP-10 levels were measured in plasma supernatants by ELISA. Whole blood from three APS-1 patients and five healthy donors was also stimulated with 1,000 U/mL IFN-γ as a control. IP-10 levels were compared between the HD and APS-1 groups by implementing nonparametric Mann–Whitney tests in GraphPad Prism. Ns: not significant; **P-value < 0.001. (B) IP-10 induction after the stimulation of whole blood from five APS-1 patients and nine healthy donors with 1 ng/mL glycosylated IFN-α2, IFN-β, or IFN-ω for 16 h. IP-10 levels were measured in plasma supernatants by ELISA. Whole blood from three APS-1 patients and five healthy donors was also stimulated with 100 U/mL IFN-γ as a control. Multiple Mann–Whitney tests were performed to compare the HD and APS-1 groups for each set of stimulation conditions. IP-10 levels were compared between the HD and APS-1 groups in nonparametric Mann–Whitney tests implemented in GraphPad Prism. Ns: not significant; **P-value < 0.001. (C) IP-10 induction in plasma supernatants after the stimulation of whole blood from a patient neutralizing low concentrations of IFN-α2 (1 ng/mL to 100 pg/mL) in a luciferase assay. Whole-blood samples from the patient and three healthy donors were stimulated with glycosylated type I (10 ng/mL) or type II (1,000 U/mL) IFNs for 16 h. IP-10 levels were then assessed in plasma supernatants by ELISA. (D) IP-10 induction in plasma supernatants after the stimulation of whole blood from a patient neutralizing low concentrations of IFN-α2 (1 ng/mL to 100 pg/mL) in a luciferase assay. Whole-blood samples from the patient and three healthy donors were stimulated with glycosylated type I (1 ng/mL) or type II (100 U/mL) IFNs for 16 h. IP-10 levels were then assessed in plasma supernatants by ELISA. (E) IP-10 induction in plasma supernatants after the stimulation of whole blood from a patient with a NFKB2 p52LOF/IκBδGOF mutation, a female patient with NEMO deficiency (incontinentia pigmenti), and a patient with auto-Abs neutralizing type I IFNs with no genetic diagnosis. TC: Travel control. Whole-blood samples were stimulated with glycosylated type I (1 ng/mL) or type II (100 U/mL) IFNs for 16 h. IP-10 levels were then assessed in plasma supernatants by ELISA. (F) IP-10 induction in plasma samples from patients with autosomal recessive IFNAR1, IFNAR2, TYK2, or IRF9 deficiency. Whole-blood samples from the patients and three healthy donors were stimulated with glycosylated type I (1 ng/mL) or type II (100 U/mL) IFNs for 16 h. IP-10 levels were then assessed in plasma supernatants by ELISA. A TC blood sample from a healthy individual was available for one of the IFNAR2−/− patients. These samples were stimulated >48 h after blood sampling. The blood of the TYK2−/− patient was stimulated 24 h after blood sampling. The blood of the IRF9−/− patient was stimulated 6 h after blood sampling.