Journal: eBioMedicine

Article Title: Autoantibodies against type I interferons in patients with zoonotic H7N9 influenza: an observational case–control study

doi: 10.1016/j.ebiom.2026.106387

Figure Lengend Snippet: Autoantibodies binding to IFNα 2 , IFNβ 1b and IFNω in patients with H7N9 infection and healthy controls. (A) Age and sex distribution of the three study groups. For each age group, the number of individuals positive for autoantibodies neutralising at least one tested IFN-I (IFNα 2 , IFNβ 1b , IFNω) at low concentrations is indicated in red. M, male; F, female; nAb+, positive for IFN-I-neutralising autoantibodies. (B) Detection of IgG autoantibodies binding to IFNα 2 , IFNβ 1b or IFNω in serum samples by multiplex bead-based assay. Samples with a Z-score >7 were considered positive for IFN-I-binding autoantibodies. Measurements were performed without technical replicates because of limited sample availability. (C) Prevalence of IFN-I-binding autoantibodies by IFN type and study group. IFNα 2 ± IFNβ 1b ± IFNω, positive for autoantibodies binding to at least one of the tested IFN-I; IFNα 2 + IFNω, positive for autoantibodies binding to both IFNα 2 and IFNω.

Article Snippet: Serum was diluted 1:50 in DMEM supplemented with 10% FCS and penicillin/streptomycin, and pre-incubated for 1 h at room temperature with one of the following IFN concentrations: (i) IFNα 2 (Novus Biologicals, NBP2-34971) at 10 ng/ml or 0.5 ng/ml; (ii) IFNβ 1b (PBL Assay Science, 11420-1) at 1 ng/ml or 0.25 ng/ml; (iii) IFN-ω (Novus Biologicals, NBP2-35893) at 10 ng/ml or 0.2 ng/ml.

Techniques: Binding Assay, Infection, Multiplex Assay, Bead-based Assay

Journal: eBioMedicine

Article Title: Autoantibodies against type I interferons in patients with zoonotic H7N9 influenza: an observational case–control study

doi: 10.1016/j.ebiom.2026.106387

Figure Lengend Snippet: Autoantibodies neutralising IFN-I in patients with H7N9 infection and healthy controls. (A) Luciferase-based reporter assay to assess the capacity of autoantibody positive sera to neutralise IFNα 2 (10 or 0.5 ng/ml), IFNβ 1b (1 or 0.25 ng/ml) or IFNω (10 or 0.2 ng/ml). Each sample was tested in biological duplicates and the mean values are shown. Samples were classified as neutralising if the mean of the relative luciferase activities was below 25% (dotted line) of the mean of the negative pool (four autoantibody-negative control sera). All sera positive for IFN-I-binding autoantibodies were tested; numbers are indicated above the graphs. Lines connect measurements of neutralising activity from the same serum sample at low and high IFN concentrations. (B) Prevalence of autoantibodies neutralising low IFN concentrations (IFNα 2 : 0.5 ng/ml, IFNβ 1b : 0.25 ng/ml; IFNω: 0.2 ng/ml) by IFN type and study group. IFNα 2 ± IFNβ 1b ± IFNω, positive for autoantibodies neutralising at least one tested IFN-I; IFNα 2 + IFNω, positive for autoantibodies neutralising both IFNα 2 and IFNω. (C) Area-proportional Venn diagrams illustrating the absolute numbers of samples with autoantibodies neutralising high and low concentrations of IFNα 2 (10 or 0.5 ng/ml), IFNβ 1b (1 or 0.25 ng/ml) or IFNω (10 or 0.2 ng/ml). Venn diagrams were created with BioVenn ( https://www.biovenn.nl/index.php ).

Article Snippet: Serum was diluted 1:50 in DMEM supplemented with 10% FCS and penicillin/streptomycin, and pre-incubated for 1 h at room temperature with one of the following IFN concentrations: (i) IFNα 2 (Novus Biologicals, NBP2-34971) at 10 ng/ml or 0.5 ng/ml; (ii) IFNβ 1b (PBL Assay Science, 11420-1) at 1 ng/ml or 0.25 ng/ml; (iii) IFN-ω (Novus Biologicals, NBP2-35893) at 10 ng/ml or 0.2 ng/ml.

Techniques: Infection, Luciferase, Reporter Assay, Negative Control, Binding Assay, Activity Assay

Journal: eBioMedicine

Article Title: Autoantibodies against type I interferons in patients with zoonotic H7N9 influenza: an observational case–control study

doi: 10.1016/j.ebiom.2026.106387

Figure Lengend Snippet: Association between the presence of IFN-I-neutralising autoantibodies and H7N9 infection. (A) The association between age, sex and IFN-I-neutralising autoantibodies in patients with H7N9 infection or in the two control groups combined (poultry workers + close contacts) was assessed using Firth's penalised logistic regression. Predicted probabilities for the presence of autoantibodies with 95% confidence intervals (CIs, shaded areas around the curve) are shown across participant age for men and women. To visualise the modelled probabilities in relation to the underlying data, we overlaid sex-specific age density distributions beneath the predicted probability curves. (B) Odds ratios (OR) with 95% CIs for the presence of autoantibodies neutralising low IFN concentrations in patients compared to healthy controls, adjusted for age and sex, determined by Firth’s penalised logistic regression models. See also for the results of the logistic regression analyses and for unadjusted estimates. IFNα 2 ± IFNω ± IFNβ 1b , positive for autoantibodies neutralising at least one of the tested IFN-I; IFNα 2 ± IFNω, positive for autoantibodies neutralising IFNα 2 and/or IFNω; ∗∗∗∗, p < 0.0001 (Firth’s penalised logistic regression).

Article Snippet: Serum was diluted 1:50 in DMEM supplemented with 10% FCS and penicillin/streptomycin, and pre-incubated for 1 h at room temperature with one of the following IFN concentrations: (i) IFNα 2 (Novus Biologicals, NBP2-34971) at 10 ng/ml or 0.5 ng/ml; (ii) IFNβ 1b (PBL Assay Science, 11420-1) at 1 ng/ml or 0.25 ng/ml; (iii) IFN-ω (Novus Biologicals, NBP2-35893) at 10 ng/ml or 0.2 ng/ml.

Techniques: Infection, Control

Journal: eBioMedicine

Article Title: Autoantibodies against type I interferons in patients with zoonotic H7N9 influenza: an observational case–control study

doi: 10.1016/j.ebiom.2026.106387

Figure Lengend Snippet: Neutralising sera block the antiviral effect of IFNα 2 in cell culture infected with IAV. Antiviral activity of IFNα 2 (5 ng/ml) against IAV (PR8-GFP, MOI 1) alone or in the presence of serially diluted IFN-I-neutralising sera (n = 19), autoantibody-negative sera (n = 4), or a monoclonal anti-IFNα 2 antibody in A549 cells. Infection rates (GFP + /DAPI + cells) at 7 h post-infection were normalised to untreated, infected cells. The dotted line indicates the reduction of infected cells after IFN treatment alone. If possible, the mean of two independent experiments is shown. Sufficient material was available for 12 out of 19 samples.

Article Snippet: Serum was diluted 1:50 in DMEM supplemented with 10% FCS and penicillin/streptomycin, and pre-incubated for 1 h at room temperature with one of the following IFN concentrations: (i) IFNα 2 (Novus Biologicals, NBP2-34971) at 10 ng/ml or 0.5 ng/ml; (ii) IFNβ 1b (PBL Assay Science, 11420-1) at 1 ng/ml or 0.25 ng/ml; (iii) IFN-ω (Novus Biologicals, NBP2-35893) at 10 ng/ml or 0.2 ng/ml.

Techniques: Blocking Assay, Cell Culture, Infection, Activity Assay

Journal: The Journal of Experimental Medicine

Article Title: Interferon-λ drives renal fibrosis by coordinating epithelial–fibroblast crosstalk

doi: 10.1084/jem.20251858

Figure Lengend Snippet: Type I and II IFNs differ from type III IFN in their regulation of TGF-β expression and the ERK–JNK pathway in renal fibroblasts during kidney fibrosis. (A–C) Primary renal fibroblasts from WT mice were treated for 24 h with 100 ng/ml of various IFNs subtypes (IFN-α, IFN-β, IFN-γ, or IFN-λ2) or PBS. (A and B) Isg15 , Mx1 , Ifit1 , and Tgf-β mRNA levels were assessed by RT-qPCR ( n = 6), and (C) TGF-β protein in the culture supernatants was quantified by ELISA ( n = 4). (D–F) Primary skin fibroblasts from WT mice were treated for 24 h with 100 ng/ml of various IFNs subtypes (IFN-α, IFN-β, IFN-γ, or IFN-λ2) or PBS. (D and E) Isg15 , Mx1 , Ifit1 , and Tgf-β mRNA levels were detected by RT-qPCR ( n = 6), and (F) TGF-β protein in the culture supernatants was measured by ELISA ( n = 4). (G–J) WT, Ifnar –/– , and Ifngr1 −/− mice were subjected to sham or UUO surgery, and kidneys were collected on day 7. n = 6 per group. (G) Representative images and quantitative analysis of fibrotic areas with Masson’s trichrome and PSR staining (scale bars = 50 μm). (H) RT-qPCR analysis of Acta2 , fibronectin, and vimentin mRNA levels in kidneys. TGF-β mRNA and protein levels in kidneys were measured by RT-qPCR (I) and western blot (J). (K and L) Primary kidney fibroblasts were treated with 100 ng/ml IFN-α (K) or IFN-β (L) for the indicated times. (K and L) Western blot analysis of phosphorylated and total ERK and JNK protein levels. Data in A–I are pooled from two independent experiments. Data in J–L are representative of three independent experiments. Data are presented as mean ± SEM. *P < 0.05, ****P < 0.0001, by two-way ANOVA with Tukey’s multiple-comparison test (A–I). ns, no significant difference. Source data are available for this figure: .

Article Snippet: Primary renal fibroblasts isolated from WT and Ifnlr1 −/− mice were stimulated with or without 100 ng/ml IFN-λ2 (250-33; PeproTech) for 1 h or 24 h. In separate experiments, primary renal and skin fibroblasts from WT mice were treated with 100 ng/ml of IFN-λ2 (250-33; PeproTech), IFN-α (CK83; Novoprotein), IFN-β (HY- P73130 ; MedChemExpress), or IFN-γ (315-05; PeproTech) for different times.

Techniques: Expressing, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Staining, Western Blot, Comparison