Panel A shows the percentage of interferon gamma-positive mucosal-associated invariant T cells after stimulation with interleukin-23 and interleukin-18. The horizontal axis represents different stimulation conditions, including no stimulation, interleukin-18, interleukin-23, and interleukin-18 plus interleukin-23, while the vertical axis represents the percentage of interferon gamma-positive cells. Panel B shows the percentage of interferon gamma-positive V delta 2 T cells under the same stimulation conditions. The horizontal axis represents the different stimulation conditions, and the vertical axis represents the percentage of interferon gamma-positive cells. Panel C shows the percentage of interferon gamma-positive natural killer cells. The horizontal axis represents the different stimulation conditions, and the vertical axis represents the percentage of interferon gamma-positive cells. Panel D shows the fold change of interferon gamma production in peripheral blood mononuclear cells after stimulation with interleukin-23, interleukin-1 beta, or interleukin-23 plus interleukin-1 beta. The horizontal axis represents the stimulation conditions, and the vertical axis represents the fold change in interferon gamma production. Panel E shows the production of interferon gamma and tumor necrosis factor after stimulation of peripheral blood mononuclear cells with phorbol 12-myristate 13-acetate and ionomycin. The horizontal axis represents the different conditions, and the vertical axis represents the concentration of the cytokines. Panel F shows the production of interferon gamma and tumor necrosis factor after interleukin-23 stimulation of peripheral blood mononuclear cells. The horizontal axis represents the different conditions, and the vertical axis represents the concentration of the cytokines. Each panel includes data from different genotypes, distinguished by different colors and symbols.
Impaired IFN-γ induction in PBMCs from individuals homozygous for hypomorphic IL23R variants. (A–C) Freshly thawed PBMCs of the indicated genotypes were stimulated with IL-23 (100 ng/ml) in the presence or absence of IL-18 (25 ng/ml) for 48 h. Percentages of IFN-γ+ cells were assessed: (A) among MAIT cells, (B) among Vδ2+ γδ T cells, (C) among NK cells. Percentages of IFN-γ+ and TNF+ cells after PMA/ionomycin were evaluated as controls and are shown in Fig. S5, G–I. Healthy (n = 16) controls, IL23RG149R/G149R (n = 1), IL23RG300V/G300V (n = 5), IL23RR381Q/R381Q (n = 6), IL12Rβ1−/− (n = 1), and TYK2P1104A/P1104A (n = 5) individuals were investigated. Nonparametric Mann–Whitney U tests were used for analysis, with *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001. (D) Fresh PBMCs of the indicated genotypes were left unstimulated (NS) or were stimulated with IL-23, IL-1β, or both for 48 h, or with PMA + ionomycin for 24 h. IFN-γ levels in the supernatants were assessed by LEGENDplex multiplex ELISA. Data were normalized against levels in the absence of stimulation. Healthy (n = 21) controls, IL23RG149R/G149R (n = 2), IL23RG300V/G300V (n = 2), IL23RR381Q/R381Q (n = 11), IL12Rβ1−/− (n = 1), and IL23R−/− (n = 3) individuals were investigated. Nonparametric Mann–Whitney U tests were used for analysis, with *P < 0.05, **P < 0.01, and ***P < 0.001. (E and F) The production of IFN-γ and TNF after PMA-ionomycin stimulation was monitored as a control, as shown in Fig. 4, D and E.