il 1β  (R&D Systems)

Journal: Stem Cell Research & Therapy

Article Title: IL-1β modulates inflammatory response of human bone marrow-derived MSCs and neutrophil recruitment in vitro via NF-kB-associated signaling

doi: 10.1186/s13287-026-05029-x

Figure Lengend Snippet: IL-1β stimulation changes the transcriptomic profile of BM-hMSCs. Schematic of experimental design A . BM-hMSCs were exposed to IL-1 β (3 replicates, 1 donor) or left unstimulated (3 replicates, 1 donor) for one hour, followed by bulk RNA sequencing. PCA plot was used to show the variance between the unstimulated control (green) and IL-1 β exposed (purple) samples B . Heatmap displaying the Z-score of the top 100 varying genes across all samples. Red color indicates higher expression of the genes and blue color indicates decreased expression C . Volcano plot of differentially expressed genes, showing their log2 fold change (X-axis) and -log10 adjusted p-values (Y-axis) D . IL-1 β Interleukin-1β, BM-hMSCs, Bone marrow derived human mesenchymal cells, PCA Principal component analysis, FC Fold change, NS not significant. Figure 1A was created using Biorender.com

Article Snippet: Following synchronization, medium was removed, and cells were stimulated with IL-1β (20 ng/ml in DPBS with 0.1% Bovine Serum Albumin, cat# 208-IL-010, R&D Systems) for 1 h (RNA-sequencing) or 24 h. Unstimulated control BM-hMSCs were exposed to serum free DMEM only.

Techniques: RNA Sequencing, Control, Expressing, Derivative Assay

Journal: Stem Cell Research & Therapy

Article Title: IL-1β modulates inflammatory response of human bone marrow-derived MSCs and neutrophil recruitment in vitro via NF-kB-associated signaling

doi: 10.1186/s13287-026-05029-x

Figure Lengend Snippet: IL-1β stimulation increases expression of neutrophil recruitment and NF-kB signaling genes by BM-hMSCs. Heatmap clustering of the Z-score of the top 20 differentially expressed genes between unstimulated control BM-hMSCs (3 replicates, 1 donor, green) and IL-1 β stimulated BM-hMSCs (3 replicates, 1 donor, purple). Red color indicates higher expression of the genes and blue color indicates decreased expression A . Protein expression of CXCL1 (10 replicates, 4 donors), CCL2 (4 replicates, 2 donors), CXCL5 (4 replicates, 2 donors), and CXCL8/IL-8 (7 replicates, 3 donors) in BM-hMSCs secretome measured by ELLA or ELISA. Data are presented as median, and statistical analysis was performed using unpaired t-test with Welch’s correction B . Bar plot of the top 20 significant Gene Ontology (GO) biological processes representing IL- 1β induced upregulated differentially expressed genes C IL-1 β, Interleukin-1β; BM-hMSCs, Bone marrow derived human mesenchymal cells; CCL2, chemokine (C–C motif) ligand 2; CXCL1, chemokine (C-X-C motif) ligand 1; CXCL2, chemokine (C-X-C motif) ligand 1; CXCL8, chemokine (C-X-C motif) ligand 8; ****, p < 0,0001

Article Snippet: Following synchronization, medium was removed, and cells were stimulated with IL-1β (20 ng/ml in DPBS with 0.1% Bovine Serum Albumin, cat# 208-IL-010, R&D Systems) for 1 h (RNA-sequencing) or 24 h. Unstimulated control BM-hMSCs were exposed to serum free DMEM only.

Techniques: Expressing, Control, Enzyme-linked Immunosorbent Assay, Derivative Assay

Journal: Stem Cell Research & Therapy

Article Title: IL-1β modulates inflammatory response of human bone marrow-derived MSCs and neutrophil recruitment in vitro via NF-kB-associated signaling

doi: 10.1186/s13287-026-05029-x

Figure Lengend Snippet: IL-1β stimulated BM-hMSCs increased neutrophil recruitment partly via the NF-kB signaling pathway. Schematic of experimental design of the neutrophil migration assay. Illustration was created with BioRender.com A . Number of neutrophils that migrated through the transwell membrane from the top well to the bottom towards control conditioned medium, conditioned medium from IL-1β stimulated BM-hMSCs, and conditioned medium from IL-1β with the NF-kB inhibitor stimulated BM-hMSCs. The experiment was performed using neutrophils from four different donors (each donor represent one data point) and the experiments were performed on three different days. B . Protein expression of CXCL1 was measured in conditioned medium from unstimulated BM-hMSCs (4 replicates, 1 donor, green), IL-1β stimulated BM-hMSCs (4 replicates, 1 donor, purple), and ILβ stimulated BM-hMSCs with 10 μM BAY 11–7082 (4 replicates, 1 donor, grey) C . Phospho-p65 levels were quantified and normalized to total p65. Data are expressed relative to control, which was set to 1 D . IL-1β Interleukin-1β BM-hMSCs Bone marrow-derived human mesenchymal cells, Ctrl control, CXCL1 chemokine (C-X-C motif) ligand 1, ns not significant; * p < 0,05; ****, p < 0,0001

Article Snippet: Following synchronization, medium was removed, and cells were stimulated with IL-1β (20 ng/ml in DPBS with 0.1% Bovine Serum Albumin, cat# 208-IL-010, R&D Systems) for 1 h (RNA-sequencing) or 24 h. Unstimulated control BM-hMSCs were exposed to serum free DMEM only.

Techniques: Migration, Membrane, Control, Expressing, Derivative Assay

Journal: Respiratory Research

Article Title: LL-37 and citrullinated-LL-37 enhances oxylipins: citrullination attenuates LL-37-mediated COX-2-dependent chemokine response in human bronchial epithelial cells

doi: 10.1186/s12931-026-03493-w

Figure Lengend Snippet: Correlations between LL-37 and citLL-37-mediated increase in the mRNA abundance of COX-2 and chemokines. HBEC-3KT cells were stimulated with either (⬤) LL-37 or (🞅) citLL-37 (0.50 µM each). mRNA abundance of COX-2, IL-8, GROα and MIP-3α were examined in cell lysates using qRT-PCR after 4 h. Relative fold changes were calculated compared to unstimulated cells normalized to 1, using the ΔΔCt method after normalization with 18s RNA expression. Pearson’s correlation analysis was performed to determine the correlation between COX-2 and ( a ) IL-8, ( b ) GROα and ( c ) MIP-3α mRNA abundance (fold changes compared to unstimulated cells)

Article Snippet: Human recombinant IL-8 (carrier-free) was purchased from R&D Systems (Oakville, ON, CA; Cat# 208-IL), aliquoted and stored at -80 °C until use.

Techniques: Quantitative RT-PCR, RNA Expression

Journal: Respiratory Research

Article Title: LL-37 and citrullinated-LL-37 enhances oxylipins: citrullination attenuates LL-37-mediated COX-2-dependent chemokine response in human bronchial epithelial cells

doi: 10.1186/s12931-026-03493-w

Figure Lengend Snippet: Inhibition of COX-2 suppresses LL-37-mediated chemokine production. HBEC-3KT cells were pre-treated with COX-2 inhibitor Rofecoxib (20 nM) for 1 h, followed by stimulation with either LL-37, citLL-37 or sLL-37 (0.50 µM each) for 24 h (n = 5). TC supernatants were examined for the abundance of ( a ) IL-8, ( b ) GROα and ( c ) MIP-3α by ELISA. Chemokine levels shown in pg/mL after background subtraction of levels in unstimulated cells. Each dot represents an independent experiment (n = 5), the bars show IQR with the median line and the whiskers represent the min-max range. Statistical significance was measured using Two-Way ANOVA, and # represents statistical significance compared to unstimulated cells ( ** p < 0.005, *** p < 0.0005, #### or **** p < 0.0001)

Article Snippet: Human recombinant IL-8 (carrier-free) was purchased from R&D Systems (Oakville, ON, CA; Cat# 208-IL), aliquoted and stored at -80 °C until use.

Techniques: Inhibition, Enzyme-linked Immunosorbent Assay

Journal: Respiratory Research

Article Title: LL-37 and citrullinated-LL-37 enhances oxylipins: citrullination attenuates LL-37-mediated COX-2-dependent chemokine response in human bronchial epithelial cells

doi: 10.1186/s12931-026-03493-w

Figure Lengend Snippet: LL-37-mediated neutrophil migration is dependent on the COX-2 pathway. a HBEC-3KT cells and ( b ) human primary bronchial epithelial cells (PBEC) were pre-treated with the COX-2 inhibitor Rofecoxib (20 nM) for 1 h, followed by stimulation with either LL-37, citLL-37 or sLL-37 (0.50 μM each). TC supernatants collected after 24 h were used in the bottom chamber of a Transwell plates. IL-8 (30 ng/mL) was used in the bottom chamber of an independent well as a positive control for neutrophil migration. Neutrophils isolated from human blood, from two independent donors with two technical replicates each, were placed in the upper chamber of the Transwell plate. Neutrophils were counted in the bottom chamber of the Transwell plate after 3 h. Results shown are after subtracting background counts of neutrophils in the bottom chamber in wells with TC supernatants from unstimulated cells. Each dot represents an independent replicate), the bars show IQR with the median line and the whiskers represent the min-max range. Statistical significance was measured using Two-Way ANOVA (*** p <0.001 and **** p <0.0001)

Article Snippet: Human recombinant IL-8 (carrier-free) was purchased from R&D Systems (Oakville, ON, CA; Cat# 208-IL), aliquoted and stored at -80 °C until use.

Techniques: Migration, Positive Control, Isolation

Journal: Respiratory Research

Article Title: LL-37 and citrullinated-LL-37 enhances oxylipins: citrullination attenuates LL-37-mediated COX-2-dependent chemokine response in human bronchial epithelial cells

doi: 10.1186/s12931-026-03493-w

Figure Lengend Snippet: Inhibition of PGE2 receptors (EP1-4) suppresses LL-37-mediated enhancement of chemokines. HBEC-3KT cells were pre-treated with specific inhibitors for PGE2 receptors, EP1 (SC-19220; 20 nM), EP2 (PF-044EP2; 25 nM), EP3 (L-798,106; 20 nM), or EP4 (MF498; 10 nM), 1 h prior to stimulation with either LL-37or sLL-37 (0.50 μM). Tissue culture (TC) supernatants were collected after 24 h and the abundance of ( a ) IL-8, ( b ) GROα and ( c ) MIP-3α were measured by ELISA. Each dot represents an independent experiment (n=4), the bars show IQR with the median line and the whiskers represent the min-max range. Results shown are after subtracting baseline values obtained from unstimulated cells in each independent experiment. Statistical significance was determined using Two-Way ANOVA (** p < 0.001 and **** p < 0.0001)

Article Snippet: Human recombinant IL-8 (carrier-free) was purchased from R&D Systems (Oakville, ON, CA; Cat# 208-IL), aliquoted and stored at -80 °C until use.

Techniques: Inhibition, Enzyme-linked Immunosorbent Assay

Journal: Respiratory Research

Article Title: LL-37 and citrullinated-LL-37 enhances oxylipins: citrullination attenuates LL-37-mediated COX-2-dependent chemokine response in human bronchial epithelial cells

doi: 10.1186/s12931-026-03493-w

Figure Lengend Snippet: Proposed mechanism of LL-37-COX-2 axis for chemokine production and neutrophil migration in the lungs. LL-37 engages the P2X 7 receptor in upregulating COX-2 expression which facilitates increase in the abundance of prostaglandin PGE2. Release of PGE2 may act in an autocrine manner through PGE2 receptors (EP1 − 4), resulting in enhanced production of chemokines including IL-8 and GROα, which facilitates neutrophil migration contributing to airway inflammation. However, citrullination of LL-37 dampens this pathway, potentially acting as a regulatory switch that limits the pro-inflammatory functions of LL-37 in the lungs (Figure created in BioRender.com)

Article Snippet: Human recombinant IL-8 (carrier-free) was purchased from R&D Systems (Oakville, ON, CA; Cat# 208-IL), aliquoted and stored at -80 °C until use.

Techniques: Migration, Expressing

Journal: medRxiv

Article Title: Effects of Arylsulfatase B and Pembrolizumab in Combination on Progression of Metastatic Melanoma in the B16F10 Syngeneic Mouse Model

doi: 10.64898/2026.01.02.25343295

Figure Lengend Snippet: A. In B16F10 pulmonary melanoma tissue, mRNA expression of MCP1 and IL-10 increased following treatment by Pembrolizumab or rhARSB, and the increase was enhanced by their combination. In contrast, IL-17α expression was increased by Pembrolizumab, but not by rhARSB. B. KC, the mouse analog of IL-8, and IL-6 were reduced by both rhARSB and Pembrolizumab and further reduced by their combination. In contrast, TNFα was increased by Pembrolizumab and reduced by rhARSB, with overall reduction by their combination. C. FGF2, VEGF, and EGF were reduced by ARSB, but not by Pembrolizumab, and the combination of ARSB and Pembrolizumab did not lead to additional decline. [ARSB=arylsulfatase B=N-acetylgalactosamine-4-sulfatase; EGF=epidermal growth factor; bFGF=basic fibroblast growth factor; IL=interleukin; KC=CXCL1=keratinocyte-derived cytokine; rh=recombinant human; MCP1=CCL2= monocyte chemoattractant protein-1; VEGF=vascular endothelial growth factor]

Article Snippet: Some preparations were treated with recombinant, human IL-8 (CXCL8 (1 ng/ml × 24h, 208-1L, R&D Systems).

Techniques: Expressing, Derivative Assay, Recombinant

Journal: medRxiv

Article Title: Effects of Arylsulfatase B and Pembrolizumab in Combination on Progression of Metastatic Melanoma in the B16F10 Syngeneic Mouse Model

doi: 10.64898/2026.01.02.25343295

Figure Lengend Snippet: A. Although KC expression was overall reduced by rhARSB in the mouse pulmonary metastases ( Fig.4B ), IL-8 in the spent media of A375 cells following ARSB increased (p=2.5×10 -5 ) and declined when ARSB was silenced (p=2.5×10 -5 ). In contrast, IL-8 in the cell extract declined following rhARSB declined following rhARSB (p=1.1×10 -7 ). B. PMN invasion to the A375 cells was increased by rhARSB (p=0.0041), reduced by ARSB siRNA (p=0.0026), reduced by IL-8 silencing (p=0.00027), and increased by addition of rhIL-8 (p=6.1×10 -5 ). These effects are attributable to increased IL-8 in the spent media following rhARSB. C. PMN migration from the filter on top to the bottom well was increased by IL-8 in the bottom well, but unaffected by Pembrolizumab. D. When IL-8 was silenced in the A375 cells, the effect of rhARSB + PBMC(Ac) and of Pembro + PBMC(Ac) and their combination [rhARSB+Pembro+PBMC(Ac)] on cleaved caspase-3 was reduced, consistent with an effect of IL-8 on PBMC(Ac)-mediated apoptosis. E. When PMN were added to the combination of rhARSB+Pembrolizumab+PBMC, the cleaved caspase-3 was further increased in the A375 cells (p=0.03). [Ac=activated, consi=control siRNA; IL=interleukin; rh=recombinant human; Pembro=Pembrolizumab; PBMC=peripheral blood mononuclear cells; PMN=polymorphonuclear leukocytes; si=siRNA]

Article Snippet: Some preparations were treated with recombinant, human IL-8 (CXCL8 (1 ng/ml × 24h, 208-1L, R&D Systems).

Techniques: Expressing, Migration, Control, Recombinant