Journal: International Journal of Molecular Medicine
Article Title: IL-17A + γδT cell activation via the HMGB1-TLR2/4-NF-κB signaling pathways in biliary atresia
doi: 10.3892/ijmm.2026.5852
Figure Lengend Snippet: IL-17A + γδT cells are increased and induce the inflammatory response in experimental BA. Experimental BA was induced in neonatal Balb/c mice through intraperitoneal injection of RRV; after RRV injection, (A) IL-17A levels in the liver homogenate supernatant of the murine BA model were dynamically measured through ELISA (n=5/group per time point), and (B) IL-17A + γδT cells were dynamically analyzed through flow cytometry on days 3, 7 and 14 (n=5/group per time point). (C) After knocking out the Tcrδ gene in Balb/c mice, the dynamic change in IL-17A content in the liver tissue of the Tcrδ −/− murine BA model was measured through ELISA (n=5/group per time point). (D) On day 7 of the Tcrδ −/− murine BA model, immunohistochemical staining with CK19 (upper panels, original magnification, ×100) was used to observe the morphology of intrahepatic bile ducts, and H&E staining (original magnification, ×100) was used to observe liver inflammation (middle panels) and extrahepatic bile duct morphology (lower panels) (n=5/group). (E) Incidence of BA and survival analysis in the Tcrδ −/− murine BA model (n=28 for Tcrδ −/− + RRV group, n=34 for WT + RRV group). (F-H) After adoptive transfusion of murine IL-17A + γδT cells into Tcrδ −/− mice, the aforementioned indicators were observed. (F) Dynamic changes in hepatic IL-17A levels were measured by ELISA (n=5/group per time point). (G) Liver inflammation, intrahepatic bile duct morphology and extrahepatic bile duct morphology were analyzed by immunohistochemical staining with CK19 (left panels, original magnification, ×100) and H&E staining (middle panels for liver inflammation and right panels for extrahepatic bile duct morphology, original magnification, ×100) (n=5/group). (H) Incidence of BA and survival analysis (n=32 for Tcrδ −/− + RRV + RPMI 1640 group, n=34 for Tcrδ −/− + RRV + IL-17A + γδT group). Data are presented as the mean ± standard deviation of at least three repeated experiments. *** P<0.001; ns, not significant. BA, biliary atresia; CK19, cytokeratin 19; H&E, hematoxylin and eosin; MEM, minimum essential medium; RRV, rhesus rotavirus; WT, wild-type.
Article Snippet: The Tcrδ −/− , Tlr2 −/− and Tlr4 −/− Balb/c mice (n=2 male mice and 4 female mice per genotype; age, 8-10 weeks; weight, 20-25 g) were produced by Shanghai Model Organisms Center, Inc., using a proprietary CRISPR-Cas9 gene targeting platform.
Techniques: Injection, Enzyme-linked Immunosorbent Assay, Flow Cytometry, Immunohistochemical staining, Staining, Standard Deviation