Fig. 1: Short-term exposure to IL-35 inhibits human NK cell activation and secretion of pro-inflammatory cytokines/chemokines. | Nature Communications

Fig. 1: Short-term exposure to IL-35 inhibits human NK cell activation and secretion of pro-inflammatory cytokines/chemokines.

From: Interleukin-35 impairs human NK cell effector functions and induces their ILC1-like conversion with tissue residency features

Fig. 1: Short-term exposure to IL-35 inhibits human NK cell activation and secretion of pro-inflammatory cytokines/chemokines.The alternative text for this image may have been generated using AI.

a NK cells were cultured for 24 h with IL-12, IL-15, and IL-18 (10 ng/mL), IL-2 (1000 UI/ml), and IL-35 (100 ng/ml) or their combination. Representative flow cytometry plots for CD25 and IFN-γ expression (upper) and quantification (%) (lower) of CD25+ IFN-γ+ NK cells. Mean values ± S.D are shown (n = 4 individual donors). b Representative raw histograms of IFN-γ, CD25, 4-1BB, PD-L1, LAG3, and CD69 in NK cells (left) and cumulative histogram of % positive NK cells in different culture conditions (right). Mean values ± S.D are shown (n = 5 to 6 individual donors). c Two-dimensional T-sne plots showing NK cell clustering based on surface marker profiles (IFN-γ/CD25/4-1BB/PD-L1/LAG3/CD69), after culture as in Fig. 1b. Representative T-sne (left) and quantification (%) (right) of activated NK cells. Mean values ± S.D are shown (n = 6 individual donors). d Cytokines and chemokines were quantified by ECLIA assay in supernatants from NK cells cultured for 24 h in the presence of IL-12 and IL-18 with or without IL-35. Results are expressed as fold change relative to control without IL-35. Mean values ± S.D are shown (n = 4 to 6 individual donors). e Representative flow cytometry plots (left) and quantification (right) of T-BET and EOMES expression in NK cells cultured as in Fig. 1d. Results are expressed as MFI expression in IL-35 condition relative to control without IL-35. Mean values ± S.D are shown (n = 7 individual donors). f VEGF-A, IL-8, and CCL5 were quantified by ECLIA assay in supernatants from NK cells cultured for 24 h as in Fig. 1b. Mean values ± S.D are shown (n = 3 to 6 individual donors). g Representative raw histograms (upper) and cumulative histograms (lower) of % positive NK cells for GP130, IL-12Rβ2, IL-27Rα after culture as indicated for 24 h. Mean values ± S.D are shown (n = 3 individual donors). Statistical significance was determined using paired T test. Source data are provided as a Source Data file.

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