mouse anti il 1r1  (R&D Systems)

Journal: PLOS ONE

Article Title: Tannic acid, an IL-1β-direct binding compound, ameliorates IL-1β-induced inflammation and cartilage degradation by hindering IL-1β-IL-1R1 interaction

doi: 10.1371/journal.pone.0281834

Figure Lengend Snippet: (A) 2303 natural compound libraries were tested for their ability to inhibit the interaction between recombinant human IL-1β and IL1 Receptor 1. Microtiter 96-well plates were coated with hIL-1β (100 ng/well) overnight and the blocking buffer without any single compound was used as a positive control. Diluted single compounds (20 μM) were added to each well and incubated for 2 h. After washing, recombinant human IL-1R1 (125 ng/ml) was added and incubated for 2 h. Subsequently, HRP-conjugated Anti-Human IgG Fc (1:2000) was added and incubated for 1 h. An OD450 was obtained following the TMB reaction. Data indicate mean ± SD (n = 3). (B) Dose-dependency test of selected natural compound for blocking the interaction between recombinant human IL-1β and IL-1R1. Every step was identical to primary screening except for concentrations of selected natural compound (10, 40, or 160 μM) and washing buffer (PBS containing 0.05% Tween-20 and 0.01% Triton X-100). * p <0.05 compared to the control group of IL-1β and IL1 Receptor 1 interaction without any natural compounds. (C) IL-1β-dependent HEK-Blue IL-1β cells (5×10 4 cell/well) were seeded onto a 96-well plate and treated with pre-incubation (20 min) of human IL-1β (10 ng/ml) with various concentrations of TA (0.78, 1.56, 3.125, 6.25, 12.5, 25, 50, or 100 μM). After a 24 h incubation, SEAP activity was assessed using QUANTI-Blue™ and the optical ensity (OD) at 620 nm. Cell viability was measured by analyzing the OD at 450 nm using D-Plus CCK. The IC 50 value of tannic acid was determined using the GraphPadPrism 10 software. Data indicate mean ± SD (n = 3). (D) Surface plasmon resonance (SPR) assay was used to analyze the direct binding of tannic acid to human IL-1β. Human IL-1β protein (50 μg/ml) was immobilized on a CM5 sensor chip and various concentrations of TA (1.56, 3.125, 6.25, 12.5, 25, 37.5, 50, 62.5, 75, 87.5, or 100 μΜ) were injected into the flow system with a flow rate 20 μl/min for 300 s and allowed to dissociate for 600 s. The K D values of the tannic acid against human IL-1β were obtained using the T200 BIA evaluation software.

Article Snippet: ELISA-based binding assays were performed for IL-1β-blocking candidates to block the interaction between IL-1β and IL-1R1 using 2303 natural compound libraries (TargetMol L-6000, Wellesley Hills, MA, USA; Selleckhem L-1400, Matsonford Road Radnor, PA, USA).

Techniques: Recombinant, Blocking Assay, Positive Control, Incubation, Control, Activity Assay, Software, SPR Assay, Binding Assay, Injection

Journal: PLOS ONE

Article Title: Tannic acid, an IL-1β-direct binding compound, ameliorates IL-1β-induced inflammation and cartilage degradation by hindering IL-1β-IL-1R1 interaction

doi: 10.1371/journal.pone.0281834

Figure Lengend Snippet: Human articular chondrocytes from OA patients were seeded onto 12-well plates (2×10 5 cells/well) and serum-starved cells were co-treated with various concentrations of TA (0.5, 1, or 2 μM) or IL-1R1 (1 μg/ml) and IL-1β (10 ng/ml) for 48 h. (A) The mRNA expression levels of iNOS , COX-2 , IL-6 , and TNF were measured using qRT-PCR. The relative quantity of each gene expression was normalized to the relative quantity of human GADPH. (B) Griess reaction was used to measure the NO levels in the culture supernatants and PGE2, IL-6, and TNF levels in the culture supernatants were evaluated using ELISA. # p < 0.05 compared with medium only control group and * p <0.05 compared with IL-1β-treated group.

Article Snippet: ELISA-based binding assays were performed for IL-1β-blocking candidates to block the interaction between IL-1β and IL-1R1 using 2303 natural compound libraries (TargetMol L-6000, Wellesley Hills, MA, USA; Selleckhem L-1400, Matsonford Road Radnor, PA, USA).

Techniques: Expressing, Quantitative RT-PCR, Gene Expression, Enzyme-linked Immunosorbent Assay, Control

Journal: PLOS ONE

Article Title: Tannic acid, an IL-1β-direct binding compound, ameliorates IL-1β-induced inflammation and cartilage degradation by hindering IL-1β-IL-1R1 interaction

doi: 10.1371/journal.pone.0281834

Figure Lengend Snippet: Human articular chondrocytes from OA patients were seeded onto 6-well plates (3×10 5 cells/well) and serum-starved cells were co-treated with various concentrations of TA (2 μM), IL-1R1 (1 μg/ml), or anti-IL-1 neutralizing antibody (1 μg/ml) and IL-1β (10 ng/ml) for 48 h. mRNA and protein expressions of MMPs, ADAMTSs, collagen type II, and aggrecan were detected by qRT-PCR (A) and Western blot analysis (B). The relative quantity of each gene expression was normalized to the relative quantity of human GADPH and β-actin was used as a loading control. # p < 0.05 compared with medium only control group and * p <0.05 compared with IL-1β-treated group.

Article Snippet: ELISA-based binding assays were performed for IL-1β-blocking candidates to block the interaction between IL-1β and IL-1R1 using 2303 natural compound libraries (TargetMol L-6000, Wellesley Hills, MA, USA; Selleckhem L-1400, Matsonford Road Radnor, PA, USA).

Techniques: Quantitative RT-PCR, Western Blot, Gene Expression, Control

Journal: bioRxiv

Article Title: IL-1R1 Blockade Enhances CD40 Agonist-Mediated Immune Responses but Fails to Increase Efficacy or Mitigate Hepatotoxicity in Pancreatic Cancer

doi: 10.1101/2025.02.23.639774

Figure Lengend Snippet: Mice bearing 9-day-old orthotopic PDAC tumors (confirmed by luminescence imaging) were treated every 3 days with the indicated antibodies (200 µg/mouse, intraperitoneal). (a) Bulk transcriptomic analysis of orthotopic PDAC tumors after 2 weeks of treatment initiation (*p < 0.05; unpaired t-test, n = 2 mice/group). (b) Kaplan-Meier survival curves (log-rank test, n = 5-6 mice/group). (c) Tumor growth kinetics in subcutaneous PDAC-bearing mice treated every 3 days with agonistic CD40 antibody, anti-IL-1R1 antibody, their combination, or left untreated (****p < 0.0001; two-way ANOVA, n = 5-8 mice/group). (d) Representative H&E-stained tumor sections showing necrosis after 2 weeks of treatment (n = 3-4 mice/group). (e) Quantification of necrotic areas (**p < 0.01; one-way ANOVA, n = 3-4 mice/group). (f) Flow cytometric analysis of PMN-MDSCs in peripheral blood after 2 weeks of treatment (*p < 0.05; unpaired Student’s t-test, n = 5 mice/group). (g) Volcano plot of differentially expressed genes (DEGs) in subcutaneous PDAC tumors treated with anti-IL-1R1 versus untreated controls (red: upregulated; green: downregulated; 546 up, 530 down; p ≤ 0.05, log2FC ≥ 0). (h–i) Gene Ontology (GO) enrichment analysis of DEGs from (g), showing significantly upregulated (h) and downregulated (i) biological processes. (j) Volcano plot of DEGs comparing combination therapy (CD40 + anti-IL-1R1) versus CD40 monotherapy (524 up, 534 down; p ≤ 0.05, log2FC ≥ 0). (k–l) GO enrichment analysis of DEGs from (j), highlighting upregulated (k) and downregulated (l) pathways. (m) Gene Set Enrichment Analysis (GSEA) plots demonstrating the enrichment of indicated gene sets in the combination therapy group compared to CD40 monotherapy.

Article Snippet: Beginning 9 days after tumor implantation, subcutaneous and orthotopic PDAC-bearing mice were treated with anti-IL-1R1 (Clone JAMA-147, BioXCell) or isotype control antibodies (BioXCell) administered intraperitoneally.

Techniques: Imaging, Staining

Journal: bioRxiv

Article Title: IL-1R1 Blockade Enhances CD40 Agonist-Mediated Immune Responses but Fails to Increase Efficacy or Mitigate Hepatotoxicity in Pancreatic Cancer

doi: 10.1101/2025.02.23.639774

Figure Lengend Snippet: Mice bearing 9-day-old subcutaneous PDAC tumors were treated as indicated. (a-b) Serum ALT (a) and AST (b) levels were measured 48 hours after treatment initiation using the COBAS INTEGRA 400 Plus analyzer. *p < 0.01, Wilcoxon non-parametric test; n = 6-9 mice/group. Data represent results combined from two independent experiments. (c) Representative H&E-stained liver histology images from mice collected one week after treatment initiation (n = 3 mice/group, two treatments administered). (d) Cumulative quantification of immune infiltration and histological changes (n = 3 mice/group). (e-f) Serum ALT and AST levels measured after 9 days (e) and 3 weeks (f) of treatment initiation. Combo: agonistic CD40 + anti-IL-1R1 antibody. (g-h) Transcriptomic profiling of the liver from PDAC-bearing mice one week after treatment initiation (two treatments total). KEGG pathway enrichment analysis comparing agonistic CD40 vs. no treatment (g) and combo vs. agonistic CD40 (h). The dot plot highlights pathways associated with liver function, toxicity, and immune infiltration. Dot size represents the number of genes involved (Count), while color intensity reflects statistical significance (−log10 p-value). Upregulated pathways are marked with upward triangles (▴), and downregulated pathways with downward triangles (▾). Pathways with p-value < 0.05 are considered significantly enriched.

Article Snippet: Beginning 9 days after tumor implantation, subcutaneous and orthotopic PDAC-bearing mice were treated with anti-IL-1R1 (Clone JAMA-147, BioXCell) or isotype control antibodies (BioXCell) administered intraperitoneally.

Techniques: Staining

Journal: bioRxiv

Article Title: IL-1R1 Blockade Enhances CD40 Agonist-Mediated Immune Responses but Fails to Increase Efficacy or Mitigate Hepatotoxicity in Pancreatic Cancer

doi: 10.1101/2025.02.23.639774

Figure Lengend Snippet: Mice bearing 9-day-old subcutaneous PDAC tumors were treated as indicated. Graphs show tumor growth at indicated time points. Figure: Tumor growth kinetics in mice bearing 9-day-old subcutaneous PDAC tumors treated as indicated. Tumor area (mm 2 ) was measured over 27 days following treatment initiation. Treatment groups included: no treatment (NO Tx), anti-Ly6G, agonistic CD40 (AgoCD40), AgoCD40 combined with anti-Ly6G, and AgoCD40 combined with anti-IL-1R1. Data represent mean ± SEM (n = 6-9 mice per group). Statistical significance was determined by two-way ANOVA with Tukey’s post hoc test. ***p < 0.001, ****p < 0.0001.

Article Snippet: Beginning 9 days after tumor implantation, subcutaneous and orthotopic PDAC-bearing mice were treated with anti-IL-1R1 (Clone JAMA-147, BioXCell) or isotype control antibodies (BioXCell) administered intraperitoneally.

Techniques: