tlr3 agonist  (Cell Signaling Technology Inc)

Journal: Nature Communications

Article Title: Multi-omics analysis identifies a microbiota–bile acid–TLR signaling axis driving bladder injury in interstitial cystitis

doi: 10.1038/s41467-025-68060-1

Figure Lengend Snippet: A Differentially expressed genes (DEGs) in full-thickness bladder tissues from HIC patients and controls identified via RNA sequencing (|log₂FC|≥2; adjusted p < 0.05, two-sided Wald test using DESeq2). Gene Ontology (GO) ( B ) and KEGG ( C ) enrichment analyses of DEGs (two-sided hypergeometric test with FDR correction). D UMAP visualization of single-cell transcriptomic landscape of urothelium, with urothelial cells (UCs) identified by KRT19 and UPK1A expression. GO ( E ) and KEGG ( F ) analyses of upregulated DEGs in UCs (two-sided hypergeometric test with FDR correction). GO ( G ) and KEGG ( H ) analyses of downregulated DEGs in UCs (two-sided hypergeometric test with FDR correction). I , J Gene Set Enrichment Analysis (GSEA) of altered pathways in UCs (two-sided permutation test with 1000 permutations, with multiple testing corrected using the FDR). The exact p value for TLR signaling of UCs in panel I is 0.0009. K Expression of Toll-like receptor (TLR) subtypes in UCs from HIC bladders. mRNA ( L ) and protein ( M ) expression levels of TLR3 in isolated urothelium from control and HIC patients ( n = 7 control vs. 10 HIC; data show median (IQR); two-sided Mann–Whitney U test; ns not significant; Bar for panel M: 1 cm). N Immunostaining showing the distribution of TLR3 protein in the urothelium of patients with HIC ( n = 7 control vs. 10 HIC; one section and field per patient; Bar: 100 μm). IHC immunohistochemistry, HL Hunner lesions, U urothelium, LP lamina propria (blue line), M muscularis. Source data are provided as a Source Data file.

Article Snippet: To explore the role of Toll-like receptor 3 (TLR3), TCDCA-pretreated HUCs were treated with either a TLR3 agonist (Poly(I:C) Sodium Salt, CST, Cat. 61401; 5 μg/mL) or TLR3 siRNA (Toll-like Receptor 3 siRNA I, CST, Cat. 6236; 100 nM).

Techniques: RNA Sequencing, Expressing, Isolation, Control, MANN-WHITNEY, Immunostaining, Immunohistochemistry

Journal: Nature Communications

Article Title: Multi-omics analysis identifies a microbiota–bile acid–TLR signaling axis driving bladder injury in interstitial cystitis

doi: 10.1038/s41467-025-68060-1

Figure Lengend Snippet: A Experimental workflow of E. avium colonization in antibiotic-pretreated mice (Abx-mice, n = 6 per group). Created in BioRender. Chen, J. (2025) https://BioRender.com/3lccwo1 . B Increased abundance of E. avium confirmed by metagenomic sequencing ( n = 6 per group). C Evaluation of voiding behavior following E. avium transplantation ( n = 6 per group; one measurement per mouse). D Measurement of mechanical pain threshold post-transplantation ( n = 6 per group). E Representative histological analysis of bladder tissues after E. avium transplantation ( n = 6 per group; one section and field per mouse). F , G Quantification of blood and urinary bile acids using comprehensive targeted bile acid profiling ( n = 6 per group). H Cell viability of human urothelial cells (HUCs) following TCDCA or TUDCA treatment ( n = 3 independent experiments). I Expression of ZO-1, TNF-α, and TLR3 following TCDCA (200 µM) exposure ( n = 3 independent experiments). J Experimental workflow of intravesical instillation of TCDCA in rats ( n = 5 per group). Created in BioRender. Chen, J. (2025) https://BioRender.com/3lccwo1 . K Evaluation of voiding function after TCDCA instillation ( n = 5 per group; each rate measured once). L Assessment of mechanical pain threshold following TCDCA exposure ( n = 5 per group). M Representative histology of bladder tissues post-instillation ( n = 5 per group; one section per rat, one field quantified per section). For panels ( B – H , K , L ): data are presented as median (IQR). Statistical analysis was performed using the two-sided Mann–Whitney U test. IHC immunohistochemistry, Red arrow: urothelial thinning, detachment, and exposure, Blue line: LP lamina propria, U urothelium, M muscularis. Source data are provided as a Source Data file.

Article Snippet: To explore the role of Toll-like receptor 3 (TLR3), TCDCA-pretreated HUCs were treated with either a TLR3 agonist (Poly(I:C) Sodium Salt, CST, Cat. 61401; 5 μg/mL) or TLR3 siRNA (Toll-like Receptor 3 siRNA I, CST, Cat. 6236; 100 nM).

Techniques: Sequencing, Transplantation Assay, Expressing, MANN-WHITNEY, Immunohistochemistry

Journal: Nature Communications

Article Title: Multi-omics analysis identifies a microbiota–bile acid–TLR signaling axis driving bladder injury in interstitial cystitis

doi: 10.1038/s41467-025-68060-1

Figure Lengend Snippet: A , B Expression of tight junction protein ZO-1 and inflammatory marker TNF-α after TLR3 intervention in TCDCA-pretreated human urothelial cells (HUCs) ( n = 3 independent experiments). C Transepithelial resistance (TER) changes following TLR3 intervention in TCDCA-pretreated HUCs ( n = 3 independent experiments). D , E Expression of ZO-1 and TNF-α after pentosan polysulfate sodium (PPS) administration in TCDCA-pretreated HUCs ( n = 3 independent experiments). F TER changes following PPS intervention in TCDCA-pretreated HUCs ( n = 3 independent experiments). G Experimental workflow showing TLR3 inhibitor and PPS administration to rats following intravesical TCDCA instillation ( n = 5 per group). Created in BioRender. Chen, J. (2025) https://BioRender.com/3lccwo1 . H Assessment of voiding function (one measurement per rat) and pain threshold after TLR3 inhibition ( n = 5 per group). I Representative histological images of bladder tissues post-TLR3 inhibition ( n = 5 per group; one section and field per rat). For panels ( B , C , E , F , H ): data are presented as median (IQR). Statistical analysis was performed using the Kruskal–Wallis test. ns not significant, IHC immunohistochemistry, Red arrow: urothelial thinning, detachment, and exposure, Blue line: LP lamina propria, U urothelium, M muscularis. Source data are provided as a Source Data file.

Article Snippet: To explore the role of Toll-like receptor 3 (TLR3), TCDCA-pretreated HUCs were treated with either a TLR3 agonist (Poly(I:C) Sodium Salt, CST, Cat. 61401; 5 μg/mL) or TLR3 siRNA (Toll-like Receptor 3 siRNA I, CST, Cat. 6236; 100 nM).

Techniques: Expressing, Marker, Inhibition, Immunohistochemistry

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: Innate immune pathway activated mesenchymal stromal cells improve function and histologic outcomes in a rodent osteoarthritis model

doi: 10.3389/fbioe.2025.1525969

Figure Lengend Snippet: RNA sequencing analysis comparing transcriptomes of joint tissues from mice treated with TLR3-MSC versus MSC. Volcano plot of transcriptome from n = 5 biological replicates of TLR3-MSC versus MSC treated mouse knee joints (A) . X-axis shows fold change and y-axis shows FDR adjusted p -value, with significantly upregulated genes shown as red dots and significantly downregulated genes shows as blue dots. Significance defined as FDR ≤0.05 fold change ≥2 or ≤−2. Differential gene expression (list of genes, description, unadjusted p -value and fold-change of top 20) for upregulated (B) and downregulated (C) genes in differential analysis results from TLR3-MSC versus MSC treated joints. Pathway analyses of differential gene expression were performed using normalized counts from n = 5 biological replicates of mice treated with either TLR3-MSC or MSC alone, indicating the top upregulated (D) and downregulated (E) pathways. FC, Fold Change; FDR, false discovery rate; MSC, mesenchymal stromal cells; TLR, Toll-Like receptor; STING, Stimulator of Interferon Genes.

Article Snippet: At weeks 3 and 5, MSC were trypsinized, counted, and a portion activated with the TLR3 agonist polyinosinic-polycytidylic acid (pIC) (InVivoGen, San Diego, CA) or 2′3′cGAMP (InVivoGen, San Diego, CA) to stimulate cGAS-STING pathways (stimulation dose at 10 μg/mL at a concentration of 1 × 10 6 cells/mL in growth media for 2 h stimulation time).

Techniques: RNA Sequencing, Gene Expression

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: Innate immune pathway activated mesenchymal stromal cells improve function and histologic outcomes in a rodent osteoarthritis model

doi: 10.3389/fbioe.2025.1525969

Figure Lengend Snippet: RNA sequencing analysis comparing transcriptomes of joint tissues from mice treated with STING-MSC versus TLR3-MSC. Volcano plot of transcriptome from n = 5 biological replicates of TLR3-MSC versus MSC treated mouse knee joints (A) . X-axis shows fold change and y-axis shows FDR adjusted p -value, with significantly upregulated genes shown as red dots and significantly downregulated genes shows as blue dots. Significance defined as FDR ≤0.05 fold change ≥2 or ≤−2. Venn diagram of differentially expressed genes for each comparison group and to resting MSC (B) . Differential gene expression (list of genes, description, unadjusted p -value and fold-change of top 20) for upregulated (C) and downregulated (D) genes in differential analysis results from TLR3-MSC versus MSC treated joints. Pathway analyses of differential gene expression were performed using normalized counts from n = 5 biological replicates of mice treated with either TLR3-MSC or MSC alone, indicating the top upregulated (E) and downregulated (F) pathways. FC, Fold Change; FDR, false discovery rate; MSC, mesenchymal stromal cells; TLR, Toll-Like receptor; STING, Stimulator of Interferon Genes.

Article Snippet: At weeks 3 and 5, MSC were trypsinized, counted, and a portion activated with the TLR3 agonist polyinosinic-polycytidylic acid (pIC) (InVivoGen, San Diego, CA) or 2′3′cGAMP (InVivoGen, San Diego, CA) to stimulate cGAS-STING pathways (stimulation dose at 10 μg/mL at a concentration of 1 × 10 6 cells/mL in growth media for 2 h stimulation time).

Techniques: RNA Sequencing, Comparison, Gene Expression

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: Innate immune pathway activated mesenchymal stromal cells improve function and histologic outcomes in a rodent osteoarthritis model

doi: 10.3389/fbioe.2025.1525969

Figure Lengend Snippet: Histologic representative images of mouse knees at endterm. Toluidine blue photomicrographs from (left to right) control (needle insertion alone), MSC, TLR3-MSC, and STING-MSC treated (right) limbs. Low magnification (×4) images of whole knee joints from (left to right) control (A) , MSC (D) , TLR3-MSC (G) , and STING-MSC (J) treated joints. Higher magnification (×10) images presented for evaluation of the medial compartment of (left to right) control (B) , MSC (E) , TLR3-MSC (H) , and STING-MSC (K) treated joints. Additional higher magnification image for evaluation of the lateral compartment for control (C) , MSC (F) , TLR3-MSC (I) and STING-MSC (L) treated joints. (A,D,G,J) 4x, scale bar = 200 μm; (B,C,E,F,H,I,K,L) 10x, scale bar = 20 μm. Pink arrows represent mostly cartilagenous osteophytes. The yellow circle indicates synovitis. Red arrows highlight areas of cartilage loss to below the calcified layer. White arrows outline a roughened cartilage surface, as well as fibrillation. MSC, mesenchymal stromal cells; TLR, Toll-Like receptor; STING, Stimulator of Interferon Genes.

Article Snippet: At weeks 3 and 5, MSC were trypsinized, counted, and a portion activated with the TLR3 agonist polyinosinic-polycytidylic acid (pIC) (InVivoGen, San Diego, CA) or 2′3′cGAMP (InVivoGen, San Diego, CA) to stimulate cGAS-STING pathways (stimulation dose at 10 μg/mL at a concentration of 1 × 10 6 cells/mL in growth media for 2 h stimulation time).

Techniques: Control

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: Innate immune pathway activated mesenchymal stromal cells improve function and histologic outcomes in a rodent osteoarthritis model

doi: 10.3389/fbioe.2025.1525969

Figure Lengend Snippet: OARSI histopathology scores. Scoring of histology slides was performed following DMM surgery and treatment with control (needle insertion alone), MSC, TLR3-MSC, or STING-MSC. Parameters listed include whole joint OARSI total score (A) , medial compartment total score (B) , lateral compartment total score (C) , medial tibial cartilage score (D) , medial femoral cartilage (E) , medial synovium score (F) , lateral tibial cartilage score (G) , lateral femoral cartilage score (H) , lateral synovium score (I) . Significant differences noted with p values < 0.05. MSC, mesenchymal stromal cells; TLR, Toll-Like receptor; STING, Stimulator of Interferon Genes.

Article Snippet: At weeks 3 and 5, MSC were trypsinized, counted, and a portion activated with the TLR3 agonist polyinosinic-polycytidylic acid (pIC) (InVivoGen, San Diego, CA) or 2′3′cGAMP (InVivoGen, San Diego, CA) to stimulate cGAS-STING pathways (stimulation dose at 10 μg/mL at a concentration of 1 × 10 6 cells/mL in growth media for 2 h stimulation time).

Techniques: Histopathology, Control