Suppression of type 1 cytokine-driven macrophage activation by fibrin(ogen)-αM β2 binding. WT, Fibγ390-396A, and IFNγ−/− mice were fed a control diet (AIN-93M) or an identical diet containing 0.025% ANIT for 4 weeks. (A) Hepatic macrophage number was determined by flow cytometry as described in “Methods.” Hepatic expression of mRNAs encoding the genes: (B) IFNγ and (C,F) NOS2 was determined by real-time qPCR. (D) Expression of NOS2 mRNA in BM macrophages plated on either bovine serum albumin (BSA) or surface adhered WT fibrinogen (10 μg/mL) and stimulated with recombinant mouse IFNγ and/or recombinant mouse interleukin-4 (10 ng/mL) for 24 hours. (E) Expression of NOS2 mRNA in IFNγ-stimulated BM macrophages plated on BSA, WT fibrinogen, or γ390-396A fibrinogen (10 μg/mL). (G) Serum ALT activity was determined as described in “Methods.” (H) CK-19 and (I) sirius red staining were quantified as described in “Methods.” Hepatic expression of mRNAs encoding the profibrogenic genes: (J) COL1A1, (K) ITGβ6, and (L) TGFβ2 were determined by real-time qPCR. Data are expressed as mean + SEM; n = 4 to 16 mice per group for in vivo studies; for in vitro studies, results represent macrophages from 9 mice. *P < .05 vs control diet within genotype; #P < .05 vs ANIT-exposed WT mice or respective treatment on BSA. Fib, fibrinogen.
