il 1β  (R&D Systems)

Journal: bioRxiv

Article Title: IL-1β and TNF drive endothelial dysfunction and coagulopathy in acute COVID-19

doi: 10.64898/2026.03.21.713333

Figure Lengend Snippet: (A-F) NHBE/HMVEC-L co-cultures were infected with SARS-CoV-2 (MOI 1) or mock-infected and treated with 10 mg/mL Anakinra immediately post-infection. Cells were fixed at 72h post-infection. A) Immunofluorescence staining for ICAM-1 (green), F-actin (Phalloidin; grey), and nuclei (DAPI; blue). Images are representative of n = 5 independent experiments. B) ICAM-1 intensity between conditions was analysed by one-way ANOVA, with Sidak’s multiple comparison test. C) Immunofluorescence staining for VE-cadherin (magenta), F-actin (Phalloidin; grey), and nuclei (DAPI; blue). Images are representative of n = 3 independent experiments. D) Quantification of gaps in the endothelial monolayer under different conditions was determined by calculating the percentage of the image area covered by gaps, and analysed by one-way ANOVA, with Sidak’s multiple comparison’s test. E) Zombie Red-stained cells (magenta) indicate cells (containing F-actin and nuclei) undergoing cell death. Images are representative of n = 3 independent experiments. F) The percentage of Zombie Red positive cells was analysed by one-way ANOVA, with Sidak’s multiple comparison’s test. G) IL-1β levels in the apical supernatant of NHBE monocultures infected with SARS-CoV-2 (MOI 1) or mock-infected, at 24, 48, and 72h post-infection (n = 3 independent experiments). H) TNF levels in the apical (left panel) and basal (right panel) supernatant of SARS-CoV-2-infected NHBE/HMVEC-L co-cultures, untreated or treated with Anakinra, at 24, 48, and 72h post-infection (n = 2 independent experiments). Scale bar for all images = 50 µm. 5 ROIs per experiment were quantified (small dots) and are colour-coded per experiment. The average of the 5 ROIs is represented with the large dot (colour-coded per experiment), and the data shows mean ± SEM. Asterisks indicate statistical significance: * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: HMVEC-L monocultures were stimulated with indicated concentrations of recombinant human TNF (Thermo Fisher Scientific; PHC3011), recombinant human IL-1β (RnD systems; 201-LB ) , recombinant IL-6 (Peprotech; 200-06-20UG) or recombinant human soluble IL-6R (Peprotech; 200-06RC-20UG) diluted in hydrocortisone-free 50:50 ALI/EGM-2MV medium for 24h at 37 °C, 5% CO 2 .

Techniques: Infection, Immunofluorescence, Staining, Comparison

Journal: bioRxiv

Article Title: IL-1β and TNF drive endothelial dysfunction and coagulopathy in acute COVID-19

doi: 10.64898/2026.03.21.713333

Figure Lengend Snippet: A) Representative immunohistochemistry (IHC) images of lungs from SARS-CoV-2-infected (10 4 TCID50) or mock-infected wild-type (WT), Tnf -/- , Il1b -/- , and Tnf -/- IL1b -/- mice, harvested at 3 days post-infection (dpi). Tissues were stained with CD31 (magenta), ICAM-1 (green), and DAPI (blue). Scale bar = 50 µm. Images are representative of 5 images per mouse, 3 mice per group. B) Quantification of ICAM-1 intensity in areas of CD31 staining, analysed by 2-way ANOVA with Sidak’s multiple testing correction. 5 ROIs per mouse were quantified (small dots) and are colour-coded per mouse. The average of the 5 ROIs is represented with the large dot (colour-coded per mouse), and the data shows mean ± SEM. C-F) Aged K18-hACE c57BL/6□J mice infected with SARS-CoV-2 (10 3 PFU) and treated with an isotype control or anti-IL-1β antibody at 1h or 3 days post-infection. Lungs were harvested at day 6 post-infection. C) Lung tissues were stained with CD31 (magenta), ICAM-1 (green), and DAPI (blue). Scale bar = 100 µm. Images are representative of 5 images per mouse, 4-6 mice per group. D) Quantification of ICAM-1 intensity in areas of CD31 staining, analysed by one-way ANOVA, with Sidak’s multiple comparison’s test. 5 ROIs per mouse were quantified (small dots) and are colour-coded per mouse. The average of the 5 ROIs is represented by the large dot (colour-coded per mouse), and the data show the mean ± SEM. E) Lung tissues were stained for fibrinogen. Scale bar = 50 µm. Images are representative of 4-6 mice per group. F) Quantification of fibrinogen staining intensity analysed by one-way ANOVA, with Sidak’s multiple comparison’s test. Asterisks indicate statistical significance: * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: HMVEC-L monocultures were stimulated with indicated concentrations of recombinant human TNF (Thermo Fisher Scientific; PHC3011), recombinant human IL-1β (RnD systems; 201-LB ) , recombinant IL-6 (Peprotech; 200-06-20UG) or recombinant human soluble IL-6R (Peprotech; 200-06RC-20UG) diluted in hydrocortisone-free 50:50 ALI/EGM-2MV medium for 24h at 37 °C, 5% CO 2 .

Techniques: Immunohistochemistry, Infection, Staining, Control