Journal: Biology Direct

Article Title: Single-cell analysis highlights the role of CD4 + IFN-I-related T cells in Behcet’s uveitis during Interferonα-2a therapy

doi: 10.1186/s13062-026-00810-7

Figure Lengend Snippet: Single-cell landscape of PBMCs from active BD patients (aBD), BD patients after IFNα-2a therapy (postBD) and HCs. (a) Study design. (b) UMAP plot showing the distribution of different cell clusters after annotation. (c) Expression of marker genes in different cell types. (d) Proportions of total cell clusters in 14 individual donors, including healthy donors (HC_1 to HC_4, n = 4), active BD patients (aBD_1 to aBD_4 and aBU_1 to aBU_3, n = 7), and BD patients post IFNα-2a therapy (postBD_1 to postBD_3, n = 3). (e) Proportions of total cell clusters from HCs ( n = 4), active BD ( n = 7) and postBD ( n = 3). ( f & g ) Differential abundance UMAP and beeswarm plots from PBMCs between aBD and HC groups (f) , and between postBD and aBD groups (g) by cell type. Each dot is a neighbourhood of cells calculated using miloR. Neighbourhoods that reach significance (spatial FDR < 0.05) and are coloured, by log fold-change. UMAP, uniform manifold approximation and projection

Article Snippet: The extracted PBMCs were cultured in Roswell Park Memorial Institute (RPMI)-1640 complete medium, and treated with IFNα-2a (2000 U/ml; MedChem Express, United States) for 48 h. For STAT2 siRNA knockdown assays, PBMCs were transfected with Rfect SP V2 siRNA Transfection Reagent (BioGenerator Biotechnology) according to the manufacturer’s instructions.

Techniques: Single Cell, Expressing, Marker

Journal: Biology Direct

Article Title: Single-cell analysis highlights the role of CD4 + IFN-I-related T cells in Behcet’s uveitis during Interferonα-2a therapy

doi: 10.1186/s13062-026-00810-7

Figure Lengend Snippet: Characterization of T cells in terms of enriched pathways, differential trajectories and transcription factors. (a) The average GSVA scores of some classical pathways in CD4 + IFN-I-related T cells. (b) The UMAP distribution of the AUCell score of the “Responds to interferon-α” pathway in T cells. (c) The AUCell score of the “Responds to interferon-α” pathway in different T-cell types, P < 0.05. (d) The AUCell score of the “Responds to interferon-α” pathway in CD4 + IFN-I-related T cells among the HCs, aBD and postBD patients. P < 0.05 (e & f) Pseudotime analysis showing two fates of CD4 + T-cell differentiation. (g) Significantly enriched pathways between the two fates. (h) Different transcription factors of all T-cell subtypes according to SCENIC analysis. (i) The mRNA relative expression of STAT2 in PBMCs treated with PBS, IFNα-2a, transfected with STAT2 -siRNA or control-siRNA (Each group: n = 5). (j & k) Representative flow cytometry plot and analysis proportion of CD4 + MX1 + T cells in lymphocytes in PBMC treated with PBS or IFNα-2a, and transfected with STAT2 -siRNA-1 or control-siRNA (Each group: n = 5). The data (i, j & k) are representative of five independent experiments. Median, quartiles and samples were indicated. The significance of differences was set as P < 0.05 in (i, j & k) determined by independent-samples t test

Article Snippet: The extracted PBMCs were cultured in Roswell Park Memorial Institute (RPMI)-1640 complete medium, and treated with IFNα-2a (2000 U/ml; MedChem Express, United States) for 48 h. For STAT2 siRNA knockdown assays, PBMCs were transfected with Rfect SP V2 siRNA Transfection Reagent (BioGenerator Biotechnology) according to the manufacturer’s instructions.

Techniques: Cell Differentiation, Expressing, Transfection, Control, Flow Cytometry

Journal: Biology Direct

Article Title: Single-cell analysis highlights the role of CD4 + IFN-I-related T cells in Behcet’s uveitis during Interferonα-2a therapy

doi: 10.1186/s13062-026-00810-7

Figure Lengend Snippet: The inhibitory effects of CD4 + IFN-I-related T cells on the activation of NK cells through the CD161 receptor. (a) The intensity of cell-cell interactions between CD4 + IFN-I-related T cells and other main cell types in PBMCs. (b) The result of GO enrichment based on the top 50 upregulated DEGs of NK cells in aBD patients compared with HCs. (c) The relative mRNA expression of IFN-γ and TNF-α in NK cells was measured using RT-qPCR (Each group: n = 5) (d) CD4 + T cells were stimulated with IFNα-2a (2000 U/ml) or PBS for 24 h, and the relative mRNA expression of ISG15 , MX1 and IFI6 was measured using RT-qPCR (Each group: n = 5) (e & f) Representative flow cytometry image and bar plot of the coculture system containing isolated CD4 + T cells with or without IFN-α stimulation and isolated NK cells. (Each group: n = 3) (g) The level of IFN-α in the supernatant of the coculture system containing isolated CD4 + T cells with or without IFN-α stimulation and isolated NK cells. (Each group: n = 3) (h & i) Representative flow cytometry images and bar plots in the coculture system containing isolated CD4 + T cells after IFN-α stimulation and isolated NK cells with CD161 blockage antibody or isotype antibody. (Each group: n = 3). (j) The level of IFN-α in the supernatant of coculture system containing isolated CD4 + T cells after IFN-α stimulation and isolated NK cells with CD161 blockage antibody or isotype antibody. All data are representative of at least three independent experiments. Data are represented as median and quartiles in (c) and (d) , and as means ± SEM in (f, g, i & j) . The significances of differences were set as adj. P < 0.05 in (b) and the P value in (c, d, f, g, i & j) were determined by independent-samples t- test

Article Snippet: The extracted PBMCs were cultured in Roswell Park Memorial Institute (RPMI)-1640 complete medium, and treated with IFNα-2a (2000 U/ml; MedChem Express, United States) for 48 h. For STAT2 siRNA knockdown assays, PBMCs were transfected with Rfect SP V2 siRNA Transfection Reagent (BioGenerator Biotechnology) according to the manufacturer’s instructions.

Techniques: Activation Assay, Expressing, Quantitative RT-PCR, Flow Cytometry, Isolation

Journal: bioRxiv

Article Title: Protective IFIH1 variant reduces immune-mediated islet stress and dysfunction in a type 1 diabetes genetic background

doi: 10.64898/2025.12.30.697107

Figure Lengend Snippet: (A) , Workflow schematic for treatment processing of SC-islets. (B) , Harmony UMAP of cell populations identified following scRNA-seq data integration. (C) , Heatmaps depicting the z-score of genetic marker expression per cluster for each time point. (D) , Stacked bar plots depicting the proportion of cells found in SC-islets derived from each MDA5 variant and treated with various stressors for 48 h. (E-J) , Volcano plots showing transcriptional differences when comparing (E) MDA5 946T Ctrl vs IFNα 48 h (total variables = 1920), (F) MDA5 946T Ctrl vs poly(I:C) 48 h (total variables = 769), (G) MDA5 946T Ctrl vs CVB3 48 h (total variables = 3326), (H) MDA5 627* Ctrl vs IFNα 48 h (total variables = 1700), (I) MDA5 627* Ctrl vs poly(I:C) 48 h (total variables = 1083), and (J) MDA5 627* Ctrl vs CVB3 48 h (total variables = 1815).

Article Snippet: Whole SC-islets were then treated with either endotoxin-free water (control), 50 ng/mL IFNα (R&D Systems, 10984-IF), 500 ng/mL poly(I:C) (InvivoGen, tlrl-piclv), or CVB3-Woodruff (titer = 1.98 x 10 12 pfu/mL) at a MOI of 20.

Techniques: Marker, Expressing, Derivative Assay, Variant Assay

Journal: bioRxiv

Article Title: Protective IFIH1 variant reduces immune-mediated islet stress and dysfunction in a type 1 diabetes genetic background

doi: 10.64898/2025.12.30.697107

Figure Lengend Snippet: (A-B) , Bar graphs showing the number of differentially expressed genes (DEGs) for each cell type found in (A) MDA5 946T and (B) MDA5 627* SC-islets. (C-E) , Venn diagrams comparing DEGs from both time points for SC-α, -β, and -δ cells from each MDA5 variant treated with either (C) IFNα, (D) poly(I:C), or (E) CVB3. (F-H) , Heatmaps showing log 2 fold change of DEGs unique to (F) SC-α, (G) SC-β, and (H) SC-δ cells from each MDA5 variant following stress treatments.

Article Snippet: Whole SC-islets were then treated with either endotoxin-free water (control), 50 ng/mL IFNα (R&D Systems, 10984-IF), 500 ng/mL poly(I:C) (InvivoGen, tlrl-piclv), or CVB3-Woodruff (titer = 1.98 x 10 12 pfu/mL) at a MOI of 20.

Techniques: Variant Assay

Journal: bioRxiv

Article Title: Protective IFIH1 variant reduces immune-mediated islet stress and dysfunction in a type 1 diabetes genetic background

doi: 10.64898/2025.12.30.697107

Figure Lengend Snippet: (A-B) , Heatmaps depicting log 2 fold change of (A) NFκB signaling-associated and (B) type 1 IFN-associated DEGs across SC-α, -β, and -δ cells when comparing the corresponding IFNα treatment to control or stress-treated MDA5 variants to each other. Positive fold change values (red) in variant-to-variant comparisons correspond to upregulation in MDA5 946T cells when compared to their MDA5 627* counterparts. (C-E) , rt-qPCR of type 1 IFN-associated genes (n = 3). Error bars represent s.e.m.** = Brown-Forsythe and Welch ANOVA tests followed by Dunnett’s T3 multiple comparisons test. (F) , Heatmap depicting log 2 fold chance of apoptosis-associated DEGs across SC-α, -β, and -δ cells when comparing the corresponding stress treatment to control or stress-treated MDA5 variants to each other. Positive fold change values (red) in variant-to-variant comparisons correspond to upregulation in MDA5 946T cells when compared to their MDA5 627* counterparts. (G) , Apoptosis assay measuring caspase 3/7 activity normalized to cell viability (n = 3). Error bars represent s.e.m. ** = Ordinary two-way ANOVA followed by Sidak’s multiple comparisons test. (H) , rt-qPCR of viral genome expression in CVB3-infected SC-islets (n = 3). Error bars represent s.e.m. ** = Unpaired t test. (I) , Intracellular viral titer in SC-islets following 48 h of CVB3 infection (n = 3). Error bars represent s.e.m. ** = Welch’s unpaired t test. Outlier identified in infected MDA5 627* SC-islet samples using the Grubbs’ method.

Article Snippet: Whole SC-islets were then treated with either endotoxin-free water (control), 50 ng/mL IFNα (R&D Systems, 10984-IF), 500 ng/mL poly(I:C) (InvivoGen, tlrl-piclv), or CVB3-Woodruff (titer = 1.98 x 10 12 pfu/mL) at a MOI of 20.

Techniques: Control, Variant Assay, Quantitative RT-PCR, Apoptosis Assay, Activity Assay, Expressing, Infection

Journal: Journal of Inflammation Research

Article Title: Prognostic Value of C-Reactive Protein, Procalcitonin, and Interferon-Alpha in COVID-19 and Their Association with PANoptosis: A Cohort Study in Guangxi, China

doi: 10.2147/JIR.S530980

Figure Lengend Snippet: The laboratory parameters in healthy controls and at hospital admission of patients with COVID-19 pneumonia. The plasma concentrations of IL-6 ( A ), IL-8 ( B ), IL-10 ( C ), PCT ( D ), IFN-α ( E ), IFN-β ( F ), S100A8/A9 ( G ), TRAIL ( H ), 8-OHdG ( I ), were analyzed in healthy controls (n = 53) and patients with non-severe (n = 37), and severe (n = 60) COVID-19. All data were represented as means±SD. The results of different groups were compared by employing ANOVA followed by the Dunnett post hoc test. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001.

Article Snippet: Cultured human NEUs, LYMs, and PLTs were stimulated with 5 μg/mL C-Reactive Protein from human fluids (C4063, Sigma-Aldrich), 0.65 μg/mL Recombinant Human Procalcitonin protein (ab92843, Abcam), 100 ng/mL IL-6 (HY-P7044, MCE), 100 ng/mL IL-8 (HY-P722044, MCE), 50 ng/mL IL-10 (HY-P7030, MCE) and/or 160 ng/mL IFN-α (HY-P7022, MCE) for 8 h (for PLTs and NEUs) or 24 h (for LYMs) at 37°C and 5% CO 2 .

Techniques: Clinical Proteomics

Journal: Journal of Inflammation Research

Article Title: Prognostic Value of C-Reactive Protein, Procalcitonin, and Interferon-Alpha in COVID-19 and Their Association with PANoptosis: A Cohort Study in Guangxi, China

doi: 10.2147/JIR.S530980

Figure Lengend Snippet: The predictive capacity of plasma IL-6, IL-8, IL-10, IFN-α, PCT and CRP for severity and death ( A ): Receiver operating characteristics (ROC) curves of IL-6, IL-8, IL-10, IFN-α, PCT and CRP for prediction of severity among COVID-19 patients. ( B ): Kaplan–Meier curves of overall survival (OS) for different IL-6, IL-8, IL-10, IFN-α, PCT and CRP levels in COVID-19 patients.

Article Snippet: Cultured human NEUs, LYMs, and PLTs were stimulated with 5 μg/mL C-Reactive Protein from human fluids (C4063, Sigma-Aldrich), 0.65 μg/mL Recombinant Human Procalcitonin protein (ab92843, Abcam), 100 ng/mL IL-6 (HY-P7044, MCE), 100 ng/mL IL-8 (HY-P722044, MCE), 50 ng/mL IL-10 (HY-P7030, MCE) and/or 160 ng/mL IFN-α (HY-P7022, MCE) for 8 h (for PLTs and NEUs) or 24 h (for LYMs) at 37°C and 5% CO 2 .

Techniques: Clinical Proteomics

Journal: Journal of Inflammation Research

Article Title: Prognostic Value of C-Reactive Protein, Procalcitonin, and Interferon-Alpha in COVID-19 and Their Association with PANoptosis: A Cohort Study in Guangxi, China

doi: 10.2147/JIR.S530980

Figure Lengend Snippet: Co-stimulation of CRP, PCT or IFN-α induced cell death in peripheral blood cells ( A – C ) Percentage of cell death in LYMs and PLTs after 8 h of treatment, and in NEUs after 24 h of treatment, as assessed by flow cytometry with PI staining. “Cocktail” contained all 5 pro-inflammatory biomarkers (IL-6, IL-8, IFN-α, PCT and CRP). ( D – F ) The percentage of cell death in LYMs and PLTs after 8 h of treatment, and in NEUs after 24 h of treatment with the indicated combinations. Data from three independent experiments (n=3 per group) were compared using ANOVA with Dunnett post hoc test for group comparisons. ** P < 0.01, **** P < 0.0001.

Article Snippet: Cultured human NEUs, LYMs, and PLTs were stimulated with 5 μg/mL C-Reactive Protein from human fluids (C4063, Sigma-Aldrich), 0.65 μg/mL Recombinant Human Procalcitonin protein (ab92843, Abcam), 100 ng/mL IL-6 (HY-P7044, MCE), 100 ng/mL IL-8 (HY-P722044, MCE), 50 ng/mL IL-10 (HY-P7030, MCE) and/or 160 ng/mL IFN-α (HY-P7022, MCE) for 8 h (for PLTs and NEUs) or 24 h (for LYMs) at 37°C and 5% CO 2 .

Techniques: Flow Cytometry, Staining

Journal: Journal of Inflammation Research

Article Title: Prognostic Value of C-Reactive Protein, Procalcitonin, and Interferon-Alpha in COVID-19 and Their Association with PANoptosis: A Cohort Study in Guangxi, China

doi: 10.2147/JIR.S530980

Figure Lengend Snippet: Co-stimulation of CRP, PCT, or IFN-α induced PANoptosis in peripheral blood cells ( A – C ): Western blot analysis of PANoptosis markers, including the apoptotic marker cleaved- caspase-3 (CC3), the pyroptosis marker N-GSDMD/GSDMD, and the necroptosis markers p-MLKL/MLKL, was performed in co-treatment with CRP, PCT, or IFN-α in LYMs ( A ) and PLTs ( C ) after 8 h of co-treatment, and in NEUs ( B ) after 24 h of co-treatment. ( D – F ): Immunofluorescence analysis of PANoptosis markers (cleaved-caspase3, GSDMD, and p-MLKL) and NETs biomarkers (CitH3, MPO, and HE) in co-treatment with CRP, PCT, or IFN-α in LYMs ( D ) and PLTs ( F ) after 8 h of co-treatment, and in NEUs ( E ) after 24 h of co-treatment. Magnification ×200, Scale bar = 20 μm. All data were represented as means±SD. Data from three independent experiments (n=3 per group) were compared using ANOVA with Dunnett post hoc test for group comparisons. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001.

Article Snippet: Cultured human NEUs, LYMs, and PLTs were stimulated with 5 μg/mL C-Reactive Protein from human fluids (C4063, Sigma-Aldrich), 0.65 μg/mL Recombinant Human Procalcitonin protein (ab92843, Abcam), 100 ng/mL IL-6 (HY-P7044, MCE), 100 ng/mL IL-8 (HY-P722044, MCE), 50 ng/mL IL-10 (HY-P7030, MCE) and/or 160 ng/mL IFN-α (HY-P7022, MCE) for 8 h (for PLTs and NEUs) or 24 h (for LYMs) at 37°C and 5% CO 2 .

Techniques: Western Blot, Marker, Immunofluorescence