Figure 4.

Analysis of gene expression. The induction of ISG was analyzed by real-time quantitative PCR. (A) CD14-positive monocytes from the patients (P1, P2) and two healthy controls were stimulated with 10³ IU/mL IFN-γ (for 2 or 6 h) and the induction of CXCL9 and IRF1 was investigated. The induction of CXCL9 was severely impaired in the patients’ cells, whereas the induction of IRF1 was only mildly impaired. *Differences were statistically significant in monocytes from the patients compared with those from healthy individuals (P<0.05). (B, C) The induction of CXCL9, IRF1 and ISG15 in EBV-B cells from the patients, Y701C/WT and L706S/WT cells, and cells from healthy individuals, in response to IFN-γ (B) or IFN-α (C) stimulation. EBV-B cells carrying Y701C or L706S mutations displayed severe impairment of CXCL9 induction in response to IFN stimulation. The induction of IRF1 and ISG15 was also impaired, but to a lesser extent than that of CXCL9. ISG expression was normalized with respect to GAPDH. The results shown are representative of three independent experiments, except for the CD14⁺ monocyte experiment (which was performed twice). Differences were statistically significant between cells carrying either the Y701C mutation*(P<0.05) or L706S mutation ^†(P<0.05) and EBV-B cells from healthy individuals.