Journal: eLife

Article Title: Targeted computational design of an interleukin-7 superkine with enhanced folding efficiency and immunotherapeutic efficacy

doi: 10.7554/eLife.107671

Figure Lengend Snippet: ( A ) AlphaFold validation of the first loop design version of Neo-7 (Neo-7 LDv1) using the default (left) and single sequence mode (right). ( B ) AlphaFold validation of the second loop design version of Neo-7 (left; Neo-7 LDv2) and Neo-7 LDv2 with mutations (right) favored by Rosetta fix backbone design. ( C ) Crystal structure of human IL-7 in complexation to human IL-7 receptor alpha (PDB ID = 3DI2). ( D ) Superimposition of Neo-7 structures (with or without additional disulfide bridge) predicted by AlphaFold. ( E ) Yeast display and flow cytometry validation of IL-7/Neo-7 bindings towards the IL-7 receptors. The yeast-displayed protein (different redesigned IL-7s) carries a HA-tag while the recombinant IL-7 receptors carry either a HIS tag (IL-7 receptor alpha) or a FC-tag (common-IL-2 family receptor gamma; for detection of IL-2Rγ binding, yeast cells were first incubated with recombinant IL-7Rα, washed, and subsequently incubated with IL-2Rγ.) The signal intensity of the X-axis (conferred by the binding of anti-HA mab) correlates with the expression level of the displayed protein while the signal intensity of the Y-axis (conferred by the binding of the anti-HIS/anti-FC mAb to the recombinant receptors bound to the displayed proteins) correlates with the binding affinity of the displayed proteins towards the IL-7 receptors.

Article Snippet: To assess the binding of the common IL-2 receptor γ (IL2Rγ) to neo-7 and IL7Rα complexes, the yeasts were first incubated with 50 nM of IL7Rα, washed, and incubated with 100 nM of human-Fc tagged IL-2 receptor-γ (IL2Rγ; Sino Biological Cat: 50087-M03H) at 4°C for an hour.

Techniques: Biomarker Discovery, Sequencing, Flow Cytometry, Recombinant, Binding Assay, Incubation, Expressing

Journal: Frontiers in Immunology

Article Title: Acetylsalicylic acid disrupts SARS-CoV-2 spike protein glycosylation and selectively impairs binding to ACE2

doi: 10.3389/fimmu.2025.1706997

Figure Lengend Snippet: ASA prevents S1-induced lung injury, fibrosis and inflammation in hACE2-KI mice. (A, B) Representative images of lung sections stained with H&E (A) and Masson’s trichrome (B) from mice receiving intratracheal instillation of vehicle, 15 μg S1 pre-treated overnight with vehicle (S1), 15 μg S1 pre-treated overnight with ASA 20 mg/L (ASA-treated S1) or 15 μg S1 pre-treated overnight with vehicle followed by ASA 20 mg/L administered immediately afterward through the same route (S1+ASA) at 7 days (n=3 per group). Scale bars: 100 µm for H&E and 20 µm for Masson’s trichrome staining. (C-E) Representative images and quantification of fibronectin ( C , red), MAC2 + macrophages ( D , red), and GR1 + neutrophils ( E , red) in lung tissue sections of mice receiving intratracheal instillation of vehicle, S1, ASA-treated S1 or S1+ASA at 7 days (n=3 per group). Lung structures and nuclei were counterstained with WGA lectin (green) and DAPI (blue), respectively. Scale bar: 20 µm. Data are expressed as % of fibronectin fluorescence area per high power field at ×63 magnification (% area/field) and the average number of MAC2 + or GR1 + cells per high power field at ×63 magnification. For all panels, results are shown as mean ± SEM and were analyzed with Tukey’s multiple comparison test. *p-value<0.05, **p-value<0.01, and ***p-value<0.001 vs Vehicle; °°p-value<0.01, and °°°p-value<0.001 vs S1; ## p-value<0.01, and ### p-value<0.001 vs ASA-treated S1.

Article Snippet: Then, wells were incubated with 0.1 μg/mL ACE2 with C-term human Fc tag (Invivogen, fc-hace2) for 1 hour at room temperature in PBS 1X.

Techniques: Staining, Fluorescence, Comparison, Significance Assay