Auto-Abs neutralizing type I IFNs in the patients with IFNAR1 variants. Luciferase-based neutralization assays for detecting auto-Abs neutralizing 10 ng/ml IFN-α2, IFN-ω, or IFN-β (left panel) and 100 pg/ml IFN-α2 or IFN-ω (right panel). Plasma samples from healthy controls (gray), patients with IFNAR1 variants (black; P1, 3, 6, 9, 17, and 27), and an APS-1 patient (red, positive control) were diluted 1:10. HEK293T cells were cotransfected with a plasmid containing the firefly luciferase gene under the control of an IFN-sensitive response element (ISRE)-containing promotor and a plasmid containing the Renilla luciferase gene. The cells were then treated with type I IFNs, and relative luciferase activity (RLA) was calculated by normalizing firefly luciferase activity against Renilla luciferase activity. An RLA <15% of the value for the mock treatment was considered to correspond to neutralizing activity (dotted line; Bastard et al., 2021a).
Figure S5.

Auto-Abs neutralizing type I IFNs in the patients with IFNAR1 variants. Luciferase-based neutralization assays for detecting auto-Abs neutralizing 10 ng/ml IFN-α2, IFN-ω, or IFN-β (left panel) and 100 pg/ml IFN-α2 or IFN-ω (right panel). Plasma samples from healthy controls (gray), patients with IFNAR1 variants (black; P1, 3, 6, 9, 17, and 27), and an APS-1 patient (red, positive control) were diluted 1:10. HEK293T cells were cotransfected with a plasmid containing the firefly luciferase gene under the control of an IFN-sensitive response element (ISRE)-containing promotor and a plasmid containing the Renilla luciferase gene. The cells were then treated with type I IFNs, and relative luciferase activity (RLA) was calculated by normalizing firefly luciferase activity against Renilla luciferase activity. An RLA <15% of the value for the mock treatment was considered to correspond to neutralizing activity (dotted line; Bastard et al., 2021a).

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