human il33  (R&D Systems)

Journal: Research

Article Title: Integrated Single-Cell Profiling Reveals TL1A as a Biomarker and Driver of Type 2 Inflammation via Macrophage-Dependent Immunoregulation in Asthma

doi: 10.34133/research.1190

Figure Lengend Snippet: Mechanism underlying tumor necrosis factor-like ligand 1A (TL1A)–C-C motif chemokine ligand 8 (CCL8) axis activation in the context of allergic inflammation. (A and B) Expression levels of TL1A induced by different stimuli in human airway epithelial cells or macrophages. (C) TL1A expression in uninduced macrophages, macrophages induced by interleukin 33 (IL-33), and macrophages induced by IL-33 plus interleukin 2 (IL-2). (D and E) Comparison of the expression levels of CCL8 induced by different concentrations of TL1A, quantified using reverse transcription polymerase chain reaction (RT-PCR) or transcriptomic sequencing. The messenger RNA (mRNA) levels of CCL8 after the transfection of TL1A-M plasmids, quantified using RT-PCR (F) or transcriptomic sequencing (G). (H) Death receptor 3 (DR3) knockdown abolished the elevation of CCL8 induced by TL1A stimulation. (I) CCL8 level in CD14 + macrophages treated or not with TL1A. (J) RT-PCR analysis of CCL8 transcript following DR3 small interfering RNA (siRNA) treatment. Representative data from CD14 + cells from a single donor. (K) A schematic for the administration of different signaling pathway inhibitors to investigate the pathways involved in TL1A–CCL8 axis activation (BAY11-7082, a nuclear factor kappa B [NF-κB] inhibitor; LY294002, a phosphoinositide 3-kinase [PI3K inhibitor]; PD98059, an extracellular signal-regulated kinase [ERK] inhibitor; SB203580, a p38 inhibitor; and SP600125, a c-Jun N-terminal kinase [JNK] inhibitor). Data are presented as the mean ± SD of triplicate measurements from at least 3 independent cell culture experiments. For primary human macrophage experiments (I and J), cells were derived from n = 4 healthy donors. * P < 0.05 and ** P < 0.01 compared with the respective groups. All in vitro stimulation and inhibitor treatments were analyzed in a blinded manner.

Article Snippet: The recombinant proteins, small-molecular compounds, or inhibitors were purchased as indicated: recombinant mouse IL-13, IL-33, and TSLP protein (MCE); recombinant human IL-2, IL-4, TNF-α, and IFN-γ protein (Abbkine); poly(I:C) (Selleck); lipopolysaccharide (Sigma-Aldrich); IL-1β (Thermo Fisher); and small-molecule inhibitors BAY11-7082, LY294002, PD98059, SB203580, and SP600125 (Selleck).

Techniques: Activation Assay, Expressing, Comparison, Reverse Transcription, Polymerase Chain Reaction, Reverse Transcription Polymerase Chain Reaction, Sequencing, Transfection, Knockdown, Small Interfering RNA, Cell Culture, Derivative Assay, In Vitro

Journal: Nature Communications

Article Title: Single-cell multiomics uncovers an endothelial mechanosensitive PIEZO1-IL-33 axis driving pulmonary fibrosis

doi: 10.1038/s41467-026-70193-w

Figure Lengend Snippet: A CellChat analysis of human lung snRNA-seq data. B Venn of 47 shared DEGs from four datasets ( GSE159354 , GSE132771 , GSE128033 , GSE122960 ). C Heatmap of 47-gene expression across cell types. D Correlation analysis (Pearson Correlation) of expression of IGFBP5, IL33, MGP, SPRY1, ACKR1 and mechanical stress scores in ECs. E Correlation analysis (Pearson Correlation) of expression of PIEZO1 and IGFBP5, IL33, MGP, SPRY1, ACKR1 in ECs. F Human snRNA-seq UMAP and violin of IL33 distribution and subpopulations. G Bar chart showed expression of IL33 (mean ± SEM) in IPF (n = 7966 cells) vs control (n = 4174 cells). P value was calculated using the two-tailed unpaired t-test. H Violin of IL33 in PIEZO1 - vs PIEZO1 + ECs. I Bulk-RNA-seq of saline and BLM (7 days and 14 days) induced mice (n = 3 per group). Gene expression levels were Z-score normalized. P value was calculated using the two-tailed unpaired t-test. J qPCR of Il33 in BLM mouse lung ECs. K IL-33 (green) and CD31 (red) co-staining in human IPF vs normal, quantification is shown at right (n = 10). L IL-33 (green) and CD31 (red) co-staining in Yoda1- or GsMTx4-treated fibrotic mouse lungs, quantification is shown at right (n = 3). M IL-33 (green) and CD31 (red)co-staining in Piezo1 WT and Piezo1 ΔEC BLM lungs, quantification is shown at right (n = 3). Error bars represent mean ± SEM, H, K, J, M: two-tailed unpaired t-test; L: one-way ANOVA followed Tukey’s multiple comparwasons test, All n values indicate biologically independent samples. Source data are provided as a Source Data file.

Article Snippet: The amounts of IL-33 in HUVECs supernatant were quantified using the Human IL-33 DuoSet ELISA kit (R&D Systems, #D3300B) according to the manufacturer’s instructions.

Techniques: Gene Expression, Expressing, Control, Two Tailed Test, RNA Sequencing, Saline, Staining

Journal: Nature Communications

Article Title: Single-cell multiomics uncovers an endothelial mechanosensitive PIEZO1-IL-33 axis driving pulmonary fibrosis

doi: 10.1038/s41467-026-70193-w

Figure Lengend Snippet: A Schematic of BLM-induced fibrosis in Il33 WT vs Il33 ΔEC mice. B Il33 mRNA (n = 4) and IL-33 protein in lung ECs (n = 3). C Representative Masson/PSR images and quantification (n = 5). D αSMA immunofluorescence and quantification (n = 4). E Lung hydroxyproline content (n = 5). F Schematic: EC-specific Il33 overexpression via AAV-Tie1 in Piezo1 WT and Piezo1 ΔEC mice; 4-week pretreatment, BLM at week 4, sacrifice at week 7. G IL-33 expression in ECs, immune and epithelial cells after AAV-OE- Il33 (n = 3). H Masson/PSR images and quantification (n = 5). I αSMA immunofluorescence and quantification (n = 4). J Hydroxyproline content (n = 5). Error bars: mean ± SEM; B,G: two-tailed unpaired t-test; C,D-E,I: two-way ANOVA followed Tukey’s multiple comparisons test; H,J: three-way ANOVA followed Tukey’s multiple comparisons test. All n values indicate biologically independent samples. Source data are provided as a Source Data file.

Article Snippet: The amounts of IL-33 in HUVECs supernatant were quantified using the Human IL-33 DuoSet ELISA kit (R&D Systems, #D3300B) according to the manufacturer’s instructions.

Techniques: Immunofluorescence, Over Expression, Expressing, Two Tailed Test

Journal: Nature Communications

Article Title: Single-cell multiomics uncovers an endothelial mechanosensitive PIEZO1-IL-33 axis driving pulmonary fibrosis

doi: 10.1038/s41467-026-70193-w

Figure Lengend Snippet: HUVECs under 20% stretch for 0,6,24 h or 2,12,25 kPa substrates; IL-33 secretion ( n = 3) A and mRNA (n = 3) B . C PIEZO1 knock-down with sh PIEZO1 ( n = 3). HUVECs were transduced with sh PIEZO1 for 48 h, cultured on 25 kPa substrates for 24 h or subjected to 6 h mechanical stretch before measuring IL-33 secretion ( n = 3) D and mRNA ( n = 3) E . HUVECs were cultured on 2, 12, 25 kPa substrates for 24 h or stretched (20%) for 0, 6, 24 h, followed by measurement of calpain activity F and CAPN2 protein G ( n = 3). HUVECs (sh PIEZO1 ) were cultured on 25 kPa substrates for 24 h or subjected to 6 h mechanical stretch, followed by measurement of calpain activity ( n = 3) H and CAPN2 protein levels ( n = 3) I . J CAPN2 knock-down with shCAPN2 ( n = 3). K HUVECs (sh CAPN2 ) were cultured on 25 kPa substrates for 24 h or subjected to 6 h mechanical stretch, IL-33 secretion(left) and mRNA(right) ( n = 3). L Model for paracrine IL-33 regulation. M Cistrome: STAT3 predicted to drive IL-33 regulation. N intersection of top 100 motifs in BLM-treated mouse ECs with top 10 TFs. O The protein levels of P-STAT3 and STAT3 in HUVECs treated with 20% stretch for 0 and 6 hours ( n = 3). P The protein levels of P-STAT3 and STAT3 in HUVECs (sh PIEZO1 or sh CAPN2 ) treated with 20% stretch for 6 h ( n = 3). Q STAT3 knock-down with sh STAT3 ( n = 3). R HUVECs (sh STAT3 ) were cultured on 25 kPa substrates for 24 h or subjected to 6 h mechanical stretch; IL-33 secretion(left) and mRNA(right) ( n = 3). Error bars: mean ± SEM; C , G , J , O – Q : two-tailed unpaired t-test; A , B , F : one-way ANOVA followed Šídák’s multiple comparisons test, D – E , H – I , K , R : two-way ANOVA Tukey’s multiple comparisons test. All n denote biologically independent samples. Source data are provided as a Source Data file.

Article Snippet: The amounts of IL-33 in HUVECs supernatant were quantified using the Human IL-33 DuoSet ELISA kit (R&D Systems, #D3300B) according to the manufacturer’s instructions.

Techniques: Knockdown, Transduction, Cell Culture, Activity Assay, Two Tailed Test