Journal: bioRxiv

Article Title: TLR5 drives metabolic dysfunction-associated steatohepatitis through lipid- and flagellin-induced hepatocyte injury signalling

doi: 10.64898/2026.02.05.703969

Figure Lengend Snippet: Schematic representation of precision-cut liver slices (PCLS) setup, created in Biorender. PCLS were cultured for 16 hours in the presence or absence of 1µM TH1020 (TLR5 inhibitor) and treated with bovine serum albumin (BSA) or oleic acid and palmitic acid (OA PA) for 24 hours (n=3). Representative immunohistochemistry images H&E and Collagen III quantification (% area) of PCLS shown. Scale bar 100µM and 50µM. Error bars represent mean ± standard deviation; P values determined by t test or ANOVA with post-test multiple comparisons. ANOVA, analysis of variance. Only statistically significant comparisons (p<0.05) are highlighted.

Article Snippet: FFPE tissue sections were immunostained with rabbit polyclonal antibody directed against human Collagen Type III (rabbit IgG, ref: 22734-1-A, Proteintech, 1/2000) using the fully automated BOND Max system from Leica Microsystems and a BOND Polymer Refine Detection kit (#DS9800).

Techniques: Cell Culture, Immunohistochemistry, Standard Deviation

Journal: ACR Open Rheumatology

Article Title: Lymphocyte Activation Gene 3 Regulation of Profibrotic Cytokines and Type I Collagen Production in Patients With Systemic Sclerosis

doi: 10.1002/acr2.70120

Figure Lengend Snippet: Impact of agonistic LAG‐3 antibody on proinflammatory cytokine secretion by PBMCs. (A) dcSSc (n = 8) PBMCs were prestimulated with CD3/CD28 for 24 hours in monocultures or (B) cocultured with allogeneic dcSSc fibroblasts. The cultures were treated with either an LAG‐3 Q22 agonistic antibody (LAG‐3 Ag) or an isotype control (LAG‐3 Isotype) for 48 hours. In monocultures, the agonistic LAG‐3 Ag significantly reduced the levels of IFNγ, IL‐1β, IL‐4, and TNFα. In cocultures, IFNγ, IL‐12p70, IL‐13, IL‐2, IL‐4, and TNFα were decreased significantly. In both setups, IL‐10 levels were significantly increased by the addition of agonistic LAG‐3 isotype–treated samples. Data are presented as mean with SD Q23 (* P < 0.05, ** P < 0.01). Ag, agonistic antibody; dcSSc, diffuse cutaneous systemic sclerosis; IFNγ, interferon γ; IL, interleukin; LAG‐3, lymphocyte Q24 activation gene 3; ns, not significant; NT, no treatment; PBMC, peripheral blood mononuclear cell; TNFα, tumor necrosis factor α (1425 pg/mL vs 1847 pg/mL; P = 0.001); compared to the control IgG, there was still a notable increase in IL‐10 production (8.33 pg/mL vs 6.10 pg/mL; P = 0.01). No significant changes were observed in the production of IL‐1β or IL‐6 (Figure ). Agonistic LAG‐3 antibodies suppress type I IFN production and reduce fibroblast matrix production. PBMCs from patients with dcSSc (n = 8) were stimulated with CD3/CD28 for 24 hours and cultured with or without allogeneic dcSSc fibroblasts for 48 hours. The cultures were then treated with either the LAG‐3 Ag or the isotype control (LAG‐3 Isotype). (C, D) The LAG‐3 Ag significantly reduced the production of bioactive IFNα/β in monocultures and cocultures, as measured in HEK‐Blue cells, compared to the LAG‐3 isotype control. (E) In cocultures, levels of type 1 procollagen and (F) fibronectin were significantly reduced by LAG‐3 Ag. Data are presented as mean with SD (* P < 0.05, ** P < 0.01). Ag, agonistic antibody; dcSSc, diffuse cutaneous systemic sclerosis; IFN, interferon; LAG‐3, lymphocyte activation gene 3; ns, not significant; NT, no treatment; PBMC, peripheral blood mononuclear cell.

Article Snippet: Quantification of sLAG‐3 (LAG‐3 Human Enzyme‐Linked Immunosorbent Assay [ELISA] kit # BMS2211; Invitrogen), type 1 procollagenα (Human Pro‐Collagen I alpha 1 DuoSet ELISA Cat.DY6220‐05; R&D Systems), and fibronectin (Human Fibronectin DuoSet ELISA Cat. DY1918‐05; R&D Systems) in the plasma and supernatants was performed according to the manufacturer's protocol.

Techniques: Control, Activation Assay, Cell Culture