mil 1r1 fc  (R&D Systems)

Journal: bioRxiv

Article Title: Early life γδ T cell activation enforces intestinal barrier integrity during intergenerational C. difficile colonization

doi: 10.64898/2026.04.28.721483

Figure Lengend Snippet: The early life IL-17 response is dependent on IL-1 β produced by infiltrating myeloid cells. (A) Differential abundance using MiloR analysis of single cell sequencing of total CD45+ cells of weanlings and adults. (B, C) Total number of neutrophils(B) and monocytes (C) in the colon normalized to weight of the tissue of mice at indicated ages. (D, E) Total number of neutrophils (D) and monocytes (E) in the colon lamina propria of 3-week old littermate IL-17+/-and IL17-/-mice. (F) UMAP of Il1b expression in weanling and adult colon CD45+ cells. (G) Expression of Il1b in total colon of mice at indicated ages. (H, I) Representative flow plot (H) and percent (I) of pro-IL-1β+ myeloid cells in the LP of weanling and adult colons. (J) ELISA quantification of IL-1β in the supernatant of colon explants of weanling and adult mice, normalized to weight of tissue. (K, L) Percent of IL-17+ γδ T cells (K) and percent of neutrophils (L) in colonic LP of control mice and mice treated with anti-IL-1R in vivo antibody from 2-3 weeks. (M, N) Percent of IL-17+ γδ T cells (M) and percent of neutrophils (N) in colonic LP of littermate IL-1R+/-and IL-1R-/-at 3 weeks of age. (O) UMAP of type 3 immunity score and Il1r1 expression on total thymocytes from day 0 mouse (P) Percent of IL-17+ γδ T cells in 3-week-old TCRδ-/-adoptively transferred with IL-1R+/-or IL-1R-/-thymus cells. (Q) UMAP of IL-1R1 expression in total CD45+ cells in human fetal thymus γδ T cells. (R, S ) Percent of IFNγ (R) and IL-17 (S) of Vγ9+ γδ T cells in human cord blood following stimulation with IL-1β and zoledronate. (B, C, D, E, G, I-N, P, R, and S) Each dot represents cells from an individual mouse. Data are represented as mean with SD. Graphs are pooled data from at least two independent experiments with atleast n = 3 in each experiment. (H) Contour plots are representative at least two independent experiments. Numbers indicate frequency of cells within each quadrant or box. (B, C, G) Data were analyzed using Kruskal-Wallis ANOVA test or (D, E, I-N and P) Mann-Whitney non-parametric unpaired T test (∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p <0.0001).

Article Snippet: IL-1r1-/-mice (C57BL/6, No.003245) were obtained from Jackson laboratories.

Techniques: Produced, Single Cell, Sequencing, Expressing, Enzyme-linked Immunosorbent Assay, Control, In Vivo, MANN-WHITNEY

Journal: Cell reports

Article Title: NEMO recruitment at single cytokine-receptor complexes shows quantized dynamics independent of ligand affinity

doi: 10.1016/j.celrep.2025.116637

Figure Lengend Snippet: (A) Average distance trees of IL-1β full sequences across divergent species. (B) Average distance trees across predicted receptor-ligand binding regions. The red font indicates species selected for subsequent analysis. (C) Crystal and generated structure for IL-1β (blue) in complex with the extracellular domain of IL-1R1 (orange). (D) Differences in binding conformations across generated structures of IL-1β-IL-1R1 complexes for two predicted binding interfaces, with residues colored by their estimated contributions to the change in free energy. See also .

Article Snippet: Purified, carrier-free recombinant human IL-1R1 (R&D Systems) and IL-1β orthologues (Invitrogen, R&D Systems) were resuspended in 50% (v/v) glycerol in distilled H2O for a final glycerol concentration of 12%.

Techniques: Ligand Binding Assay, Generated, Binding Assay

Journal: Cell reports

Article Title: NEMO recruitment at single cytokine-receptor complexes shows quantized dynamics independent of ligand affinity

doi: 10.1016/j.celrep.2025.116637

Figure Lengend Snippet: (A) Schematic of IL-1β-induced EGFP-NEMO complex formation at the plasma membrane. IL-1β first binds to IL-1R1, enabling recruitment of IL-1R3 to form the receptor complex. Cytoplasmic MyD88 associates with the complex, facilitating IKK recruitment and the formation of EGFP-NEMO puncta. (B) Thermal shift curves of IL-1R1 stabilized by IL-1β indicate thermal stabilization of the human receptor by indicated cytokine orthologs. (C) Melting temperatures of isolated IL-1R1 as well as IL-1R1 in complex with IL-1β orthologs, derived from thermal shift curves in (B). (D) Quantitative descriptors extracted from each single-cell time courses of EGFP-NEMO puncta. (E) Boxplots of Fano noise evaluated for single-cell time courses at each concentration for indicated species. (F) Sigmoid curves fitted to the mean values of experimental single-cell descriptors across IL-1β concentrations and species. The EC 50 is indicated. See for fit parameters. (G) EC 50 values, reflecting the concentration at which 50% of the maximal response is reached quantified from (F). (H) Stochastic simulations using a minimal model recapitulate experimental results. Simulated dose-response curves as in (D) reveal dose-response relationshipsfor each predicted affinity. See for simulation and fit parameters. (I) EC 50 values derived from simulated data, reflecting the predicted net affinity of IL-1β to form signaling-competent complexes. See for fit parameters. See also .

Article Snippet: Purified, carrier-free recombinant human IL-1R1 (R&D Systems) and IL-1β orthologues (Invitrogen, R&D Systems) were resuspended in 50% (v/v) glycerol in distilled H2O for a final glycerol concentration of 12%.

Techniques: Clinical Proteomics, Membrane, Isolation, Derivative Assay, Concentration Assay