mouse il 1β antisera  (R&D Systems)

Journal: eBioMedicine

Article Title: Interleukin-27 remodels the bone marrow niche to suppress B-cell development and leukaemia progression in mouse models

doi: 10.1016/j.ebiom.2026.106239

Figure Lengend Snippet: IL-27 impairs early B-cell development in the bone marrow. (A) Experimental scheme for AAV-mediated IL-27 delivery and analysis of splenic and BM B-cell compartments. (B–D) Splenic B-cell analysis 10 weeks after AAV-Ctrl or AAV-IL-27 treatment (n = 2 per group). (B) Frequency of total splenic B cells. (C) CIBERSORTx-based deconvolution of splenic B-cell subsets, including clustering and pseudotime analysis (left) and relative subset distribution (right). (D) Cd93 expression (TPM) in splenic B cells. (E–G) Analysis of splenic and BM B-cell compartments 3 weeks after treatment (n = 4 per group). (E) Representative flow cytometry plots. (F) Frequencies of B220 + CD19 − and B220 − CD19 + populations in the BM and spleen. (G) Relative distribution of B-cell developmental subsets in the BM. (H–I) Kinetics of B-cell populations following IL-27 treatment (n = 2 per group at each time point). (H) Frequency and absolute number of BM B220 + B cells. (I) Proportions of B-cell subsets within the B220 + compartment at the indicated time points. ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001; ns, not significant; unpaired Student's t-test.

Article Snippet: Cells were treated with recombinant murine IL-27 (51107-M08H, Sino Biological) or human IL-27 (50 ng/mL) for 24 h prior to RNA extraction.

Techniques: Cell Analysis, Expressing, Flow Cytometry

Journal: eBioMedicine

Article Title: Interleukin-27 remodels the bone marrow niche to suppress B-cell development and leukaemia progression in mouse models

doi: 10.1016/j.ebiom.2026.106239

Figure Lengend Snippet: Exogenous IL-27 inhibits CLP formation in the bone marrow. (A) qPCR analysis of B-lineage- and myeloid-associated transcription factors in BM cells 5 days after AAV-Ctrl or AAV-IL-27 administration (n = 3 per group). (B) qPCR analysis of CEBPA expression in Reh cells treated with hIL-27 and DAC for 3 days (n = 3 per group). (C–D) Frequencies and absolute numbers of CLPs in WT and CD19 Cre ;IL-27R fl/fl mice (n = 3 per group). (E–F) BM B-cell frequency (E) and proportion of B220 + CD43 hi IgM − B cells (F) in WT and CD19 Cre ;IL-27R fl/fl mice 2 weeks after AAV treatment (n = 3 for WT Ctrl group; n = 3 for WT IL-27 group; n = 3 for CD19 Cre Ctrl group; n = 4 for CD19 Cre IL-27 group). (G–H) BM B-cell frequency (G) and proportion of B220 + CD43 hi IgM − B cells (H) in CD4 Cre ;IL-27R fl/fl and Lyz2 Cre ;IL-27R fl/fl mice 2 weeks after AAV treatment (n = 3 for CD4 Cre Ctrl group; n = 3 for CD4 Cre IL-27 group; n = 5 for Lyz2 Cre Ctrl group; n = 6 for Lyz2 Cre IL-27 group). ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001; ns, not significant; unpaired Student's t-test.

Article Snippet: Cells were treated with recombinant murine IL-27 (51107-M08H, Sino Biological) or human IL-27 (50 ng/mL) for 24 h prior to RNA extraction.

Techniques: Expressing

Journal: eBioMedicine

Article Title: Interleukin-27 remodels the bone marrow niche to suppress B-cell development and leukaemia progression in mouse models

doi: 10.1016/j.ebiom.2026.106239

Figure Lengend Snippet: Exogenous IL-27 reshapes immune reconstitution after bone marrow transplantation. (A) Experimental scheme. WT mice underwent BM transplantation followed by AAV-Ctrl or AAV-IL-27 treatment. Splenic and BM immune reconstitution was analysed at 10 weeks after treatment (n = 4 mice for the Ctrl group; n = 5 mice for the IL-27 group). (B) Percentages and absolute numbers of CD45 + splenocytes. (C–F) Frequencies of major immune cell populations within CD45 + splenocytes: myeloid cells (C), macrophages (D), T and B cells (E), and NK cells (F). (G) Frequency of CD4 + T cells among splenic T cells. (H) Proportion of Tregs among CD4 + T cells. (I–J) BM B-cell reconstitution. Frequency of BM B220 + B cells (I) and distribution of B220 + B-cell subsets (J). ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001; ns, not significant; unpaired Student's t-test.

Article Snippet: Cells were treated with recombinant murine IL-27 (51107-M08H, Sino Biological) or human IL-27 (50 ng/mL) for 24 h prior to RNA extraction.

Techniques: Transplantation Assay

Journal: eBioMedicine

Article Title: Interleukin-27 remodels the bone marrow niche to suppress B-cell development and leukaemia progression in mouse models

doi: 10.1016/j.ebiom.2026.106239

Figure Lengend Snippet: IL-27 inhibits B-cell development through both direct and indirect mechanisms. (A) Schematic of the BM chimera model. WT (WTR) or IL-27R −/− (KOR) recipients were treated with AAV-Ctrl or AAV-IL-27. Donor BM cells were a 1:1 mix of WT (CD45.1 + ) and IL-27R −/− (CD45.1 − ) cells. Analyses were performed at 5 weeks (B–E; n = 3 per group) or 10 weeks (F–I; n = 5 per group) post-transplantation. (B) Percentage of CD19 + B cells from CD45.1 + or CD45.1 − (IL-27R −/− ) donors in spleen and BM of WTR and KOR mice. (C) Absolute numbers of total CD19 + B cells in spleen and BM of WTR and KOR mice. (D) Absolute numbers of CD19 + B cells from CD45.1 + or CD45.1 − (IL-27R −/− ) donors in spleen and BM of WTR and KOR mice. (E) Percentage of B220 + CD19 − B cells from CD45.1 + or CD45.1 − (IL-27R −/− ) donors in BM of WTR and KOR mice. (F) Absolute numbers of B220 + CD19 - B cells from CD45.1 + or CD45.1 - (IL-27R −/− ) donors in BM of WTR and KOR mice. (G) Frequencies and absolute numbers of BM B220 + B cells derived from CD45.1 + or IL-27R −/− donors in WTR mice. (H) Proportion of CD43 hi IgM − subsets from CD45.1 + or IL-27R −/− donors in BM B220 + B cells in WTR mice. (I–J) Frequencies of CLPs (I) and their donor-specific contributions (J) in BM chimeras. ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001; ns, not significant; unpaired Student's t-test.

Article Snippet: Cells were treated with recombinant murine IL-27 (51107-M08H, Sino Biological) or human IL-27 (50 ng/mL) for 24 h prior to RNA extraction.

Techniques: Transplantation Assay, Derivative Assay

Journal: eBioMedicine

Article Title: Interleukin-27 remodels the bone marrow niche to suppress B-cell development and leukaemia progression in mouse models

doi: 10.1016/j.ebiom.2026.106239

Figure Lengend Snippet: Exogenous IL-27 indirectly inhibits BM B-ALL cells. (A) Experimental design. GFP + B-ALL cells were transferred into WT mice, followed by AAV-Ctrl or AAV-IL-27 administration. BM and splenic B-ALL cells were analysed 9–10 days later (n = 3 per group). (B–C) Representative flow cytometry plots and quantification of GFP + B-ALL and GFP − B cells. (D) GSVA analysis of transcriptional changes in BM B-ALL cells isolated from mice in (A). (E) Gene Ontology (GO) enrichment analysis of DEGs between Ctrl and IL-27 groups in (A) (adjusted P < 0.05, |log 2 fold change| > 1). The outer ring indicates enriched GO terms, the middle ring represents relative gene expression changes in the IL-27 group, and the inner ring shows background gene counts, with colour intensity reflecting enrichment significance. Enriched pathways include interferon-γ-mediated signalling, interferon-β responses, adhesion-related processes, protozoan defence responses, calmodulin-dependent kinase signalling, and negative regulation of STAT tyrosine phosphorylation. (F–G) Expression of apoptosis- and cell cycle-related genes in BM B-ALL cells from (A). (H) Experimental design. WT and IL-27R −/− mice transplanted with GFP + B-ALL cells were treated with AAV-Ctrl or AAV-IL-27 (n = 4 for WT Ctrl group; n = 4 for WT IL-27 group; n = 5 for IL-27R −/− Ctrl group; n = 5 for IL-27R −/− IL-27 group). (I–J) Representative flow plots and quantification of BM B-ALL cells. (K–L) Flow cytometry plots and quantification of BM B-ALL cells in CD4 Cre ;IL-27R fl/fl , CD19 Cre ;IL-27R fl/fl , and Lyz2 Cre ;IL-27R fl/fl mice (n = 2 for CD4 Cre Ctrl group; n = 2 for CD4 Cre IL-27 group; n = 3 for CD19 Cre Ctrl group; n = 3 for CD19 Cre IL-27 group; n = 3 for Lyz2 Cre Ctrl group; n = 3 for Lyz2 Cre IL-27 group). (M) Flow cytometry plots and quantification of BM B-ALL cells in Rag1 −/− mice (n = 3 per group). ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001; ns, not significant; unpaired Student's t-test.

Article Snippet: Cells were treated with recombinant murine IL-27 (51107-M08H, Sino Biological) or human IL-27 (50 ng/mL) for 24 h prior to RNA extraction.

Techniques: Flow Cytometry, Isolation, Gene Expression, Phospho-proteomics, Expressing

Journal: eBioMedicine

Article Title: Interleukin-27 remodels the bone marrow niche to suppress B-cell development and leukaemia progression in mouse models

doi: 10.1016/j.ebiom.2026.106239

Figure Lengend Snippet: IL-27 reshapes the BM niche, reducing B-cell support and leukaemogenesis. (A) Pseudotime analysis of BM B-cell subsets (C0–C10). (B) Expression of representative marker genes across B-cell clusters in (A). (C–G) BM cells were harvested 8 days after AAV-Ctrl or AAV-IL-27 administration for RNA-seq analysis (n = 3 per group). (C) CIBERSORTx analysis showing increased pre-pro-B and cycling pro-B subsets after IL-27 treatment. (D) KEGG pathway enrichment analysis highlighting alterations in adhesion-related pathways. (E–F) Heatmaps of transcription factors (E) and adhesion molecules (F). (G) RNA-seq analysis of genes shown in the figure in BM cells. (H) RNA-seq analysis of genes shown in the figure in human MSCs with or without hIL-27 treatment (n = 2 per group). (I) qPCR analysis of genes shown in the figure in murine MSCs with or without IL-27 (n = 3 per group). (J) RNA-seq analysis of genes shown in the figure in BM Gr-1 + cells 3 weeks after AAV-Ctrl or AAV-IL-27 administration (n = 2 per group). ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001; ns, not significant; unpaired Student's t-test.

Article Snippet: Cells were treated with recombinant murine IL-27 (51107-M08H, Sino Biological) or human IL-27 (50 ng/mL) for 24 h prior to RNA extraction.

Techniques: Expressing, Marker, RNA Sequencing

Journal: eBioMedicine

Article Title: Interleukin-27 remodels the bone marrow niche to suppress B-cell development and leukaemia progression in mouse models

doi: 10.1016/j.ebiom.2026.106239

Figure Lengend Snippet: IL-27 exhibits therapeutic potential while modulating B-cell compartments. (A–B) Experimental design of combination therapy in WT (A) and Rag1 −/− (B) mice, with corresponding Kaplan–Meier survival analysis. (C–D) Experimental design of IL-27R −/− CAR-T cell therapy combined with AAV-Ctrl or AAV-IL-27 in B-ALL-bearing mice, with Kaplan–Meier survival curves. B-ALL cells were transplanted either prior to chemotherapy (C) or after chemotherapy (D). (E–F) NSG mice transplanted with human G-PBSCs were injected intramuscularly with AAV-Ctrl or hIL-27. 10 days later, frequencies and absolute numbers of B cells in the BM and spleen were analysed (n = 5 per group). ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001; ns, not significant; log-rank test (A–D) and unpaired Student's t-test (E–F).

Article Snippet: Cells were treated with recombinant murine IL-27 (51107-M08H, Sino Biological) or human IL-27 (50 ng/mL) for 24 h prior to RNA extraction.

Techniques: Injection

Journal: Frontiers in Pharmacology

Article Title: Thymelaea hirsuta (L.) Endl . extract attenuates NLRP3 inflammasome activation via modulation of ATPase activity

doi: 10.3389/fphar.2026.1781860

Figure Lengend Snippet: TMH inhibits activation of the NLRP3 inflammasome. (A) Schematic workflow of in vitro NLRP3 inflammasome activation. (B,C) Cell viability was assessed in J774A.1 cells (B) and PMA (500 nM)-differentiated THP-1 cells (C) treated with TMH for 2 or 18 h using the EZ-Cytox assay. (D) LPS-primed J774A.1 cells were treated with TMH for 2 h, then stimulated with ATP (5 mM) for 30 min in the presence or absence of MCC950. IL-1β (p17) levels in supernatants were measured by ELISA. (E,F) PMA-differentiated THP-1 cells were primed with LPS, treated with TMH for 2 h, then stimulated with ATP (5 mM) (E) or nigericin (10 μM) (F) for 1 h, with or without MCC950. IL-1β (p17) levels in supernatants were determined by ELISA. Data are presented as the mean ± SD (n = 3). Statistical analysis was performed using one-way ANOVA followed by Tukey’s multiple comparisons test. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001; n.s., not significant.

Article Snippet: Antibodies specific for human IL-1β (AF-401-NA) and mouse IL-1β (AF-201-NA) were purchased from R&D Systems (United States).

Techniques: Activation Assay, In Vitro, Enzyme-linked Immunosorbent Assay

Journal: Frontiers in Pharmacology

Article Title: Thymelaea hirsuta (L.) Endl . extract attenuates NLRP3 inflammasome activation via modulation of ATPase activity

doi: 10.3389/fphar.2026.1781860

Figure Lengend Snippet: TMH inhibits activation of the NLRP3 inflammasome. (A) Schematic workflow of in vitro NLRP3 inflammasome activation. (B,C) Cell viability was assessed in J774A.1 cells (B) and PMA (500 nM)-differentiated THP-1 cells (C) treated with TMH for 2 or 18 h using the EZ-Cytox assay. (D) LPS-primed J774A.1 cells were treated with TMH for 2 h, then stimulated with ATP (5 mM) for 30 min in the presence or absence of MCC950. IL-1β (p17) levels in supernatants were measured by ELISA. (E,F) PMA-differentiated THP-1 cells were primed with LPS, treated with TMH for 2 h, then stimulated with ATP (5 mM) (E) or nigericin (10 μM) (F) for 1 h, with or without MCC950. IL-1β (p17) levels in supernatants were determined by ELISA. Data are presented as the mean ± SD (n = 3). Statistical analysis was performed using one-way ANOVA followed by Tukey’s multiple comparisons test. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001; n.s., not significant.

Article Snippet: Antibodies specific for human IL-1β (AF-401-NA) and mouse IL-1β (AF-201-NA) were purchased from R&D Systems (United States).

Techniques: Activation Assay, In Vitro, Enzyme-linked Immunosorbent Assay

Journal: Frontiers in Pharmacology

Article Title: Thymelaea hirsuta (L.) Endl . extract attenuates NLRP3 inflammasome activation via modulation of ATPase activity

doi: 10.3389/fphar.2026.1781860

Figure Lengend Snippet: TMH does not affect other inflammasome or inflammatory pathways. (A,B) LPS-primed J774A.1 cells were treated with TMH for 2 h, followed by stimulation with either dsDNA (1 μg/mL) (A) or flagellin (B) transfected using Lipofectamine 2000 for 3 h. (C) Following TMH treatment (2 h), nuclear and cytosolic fractions of LPS-primed J774A.1 cells were separated and assessed for NF-κB p65 translocation by immunoblotting. (D) HEK293FT cells transfected with the pGL4.32 luciferase reporter vector were treated with TNF-α (20 ng/mL) for 5 h, with or without TMH pre-treatment (2 h). Luciferase activity was measured using the Bright-Glo™ luciferase assay system (Promega). Data are presented as the mean ± SD (n = 3). Statistical analysis was performed using one-way ANOVA followed by Tukey’s multiple comparisons test n.s., not significant.

Article Snippet: Antibodies specific for human IL-1β (AF-401-NA) and mouse IL-1β (AF-201-NA) were purchased from R&D Systems (United States).

Techniques: Transfection, Translocation Assay, Western Blot, Luciferase, Plasmid Preparation, Activity Assay

Journal: Frontiers in Pharmacology

Article Title: Thymelaea hirsuta (L.) Endl . extract attenuates NLRP3 inflammasome activation via modulation of ATPase activity

doi: 10.3389/fphar.2026.1781860

Figure Lengend Snippet: TMH inhibits NLRP3 inflammasome activation by suppressing ASC oligomerization and ATPase activity. (A) LPS-primed J774A.1 cells were treated with TMH for 2 h, then stimulated with imiquimod (200 μM) for 1 h. (B) Cells were treated with TMH (2 h), stained with MitoSOX (5 μM, 10 min), and stimulated with ATP (5 mM, 5 min). Intracellular ROS were visualized by confocal microscopy. (C,D) PMA-differentiated THP-1 cells were primed with LPS, treated with TMH for 2 h, then stimulated with nigericin (10 μM, 1 h). ASC speck formation was examined by confocal microscopy (C) , and representative images from five fields are shown (D) . (E) Cells were stimulated with or without MCC950 and cross-linked with DSS (2.5 mM, 30 min) before immunoblotting for ASC oligomerization. (F) NLRP3 ATPase activity was measured using the ADP-Glo™ assay to quantify ATP-to-ADP conversion following TMH treatment. Data are presented as the mean ± SD (n = 3). Statistical analysis was performed using one-way ANOVA followed by Tukey’s multiple comparisons test. **P < 0.01, ***P < 0.001.

Article Snippet: Antibodies specific for human IL-1β (AF-401-NA) and mouse IL-1β (AF-201-NA) were purchased from R&D Systems (United States).

Techniques: Activation Assay, Activity Assay, Staining, Confocal Microscopy, Western Blot, Glo Assay

Journal: Frontiers in Pharmacology

Article Title: Thymelaea hirsuta (L.) Endl . extract attenuates NLRP3 inflammasome activation via modulation of ATPase activity

doi: 10.3389/fphar.2026.1781860

Figure Lengend Snippet: Toxicity assessment of TMH in zebrafish embryos. (A) Survival curves of zebrafish embryos treated with TMH at concentrations of 1, 10, or 100 μg/mL. Sixty embryos were used per condition and maintained in E3 medium. The control group was treated with DMSO (0.25%, v/v), corresponding to the highest TMH concentration. (B) Survival and phenotype distribution of 5 dpf embryos treated with TMH at distinct concentrations from 1 to 5 dpf. (C) Representative images showing developmental abnormalities observed in embryos treated with 10 μg/mL TMH at 5 dpf. (D) Representative images of Sudan Black B staining in 3 dpf embryos treated with vehicle control (0.0025% DMSO with PTU; upper panel) or TMH (1 μg/mL; lower panel). (E) Quantification of neutrophil counts in the CHT (red dashed outline) following treatment with TMH (1 μg/mL). Data are presented as mean ± SEM of individual embryos. Statistical significance was determined using an unpaired two-tailed Student’s t-test; n.s., not significant (p = 0.9997). Scale bars, 300 µm.

Article Snippet: Antibodies specific for human IL-1β (AF-401-NA) and mouse IL-1β (AF-201-NA) were purchased from R&D Systems (United States).

Techniques: Control, Concentration Assay, Staining, Two Tailed Test

Journal: Frontiers in Pharmacology

Article Title: Thymelaea hirsuta (L.) Endl . extract attenuates NLRP3 inflammasome activation via modulation of ATPase activity

doi: 10.3389/fphar.2026.1781860

Figure Lengend Snippet: TMH suppresses LPS-induced inflammatory cell recruitment in zebrafish embryos. (A) Representative images of neutrophil recruitment in the CHT in 3 dpf zebrafish embryos following LPS stimulation, assessed by Sudan Black B staining. Top panel: control embryos treated with 0.0025% DMSO (n = 29); middle panel: LPS-treated embryos (10 μg/mL; n = 32); bottom panel: embryos co-treated with LPS (10 μg/mL) and TMH (1 μg/mL; n = 33). (B) Quantification of neutrophil accumulation in the CHT following LPS stimulation with or without TMH treatment using Sudan Black B staining from (A) . (C) Representative images of whole-mount in situ hybridization showing mpx -positive neutrophils in 3 dpf embryos under LPS and TMH treatment conditions. Top panel (DMSO 0.0025%; n = 34); middle panel (LPS 10 μg/mL; n = 35); bottom panel (LPS 10 μg/mL with TMH 1 μg/mL; n = 39). (D) Quantification of mpx -positive cells within the CHT from (C) . (E) Representative confocal images of macrophages in 3 dpf Tg ( mpeg1 :EGFP) zebrafish embryos following LPS and TMH treatment. Top panel (DMSO 0.0025%; n = 8); middle panel (LPS 10 μg/mL; n = 11); bottom panel (LPS 10 μg/mL with TMH 1 μg/mL; n = 13). (F) Quantification of mpeg1 :EGFP-positive macrophages in the CHT region shown in (E) . Red dashed outline indicates the CHT. All data are presented as mean ± SEM. Statistical significance was determined using an unpaired two-tailed Student’s t-test. *P < 0.05; **P < 0.01; ***P < 0.05; ****P < 0.0001. Scale bars, 300 µm.

Article Snippet: Antibodies specific for human IL-1β (AF-401-NA) and mouse IL-1β (AF-201-NA) were purchased from R&D Systems (United States).

Techniques: Staining, Control, In Situ Hybridization, Two Tailed Test