Fig. 6.

Inhibition of both transmembrane tumour necrosis factor (tmTNF) and soluble TNF (sTNF) is required to suppress interferon (IFN)-γ-induced CD40, major histocompatibility complex (MHC) class II and chemokine receptor 2 (CCR2) expression by macrophages. Bone marrow-derived macrophages (BMDMϕ) from wild-type or TNF receptor 1 (TNFR1^−/−) mice were incubated with 100 U/ml IFN-γ for 24 h prior to analysis of cell surface expression of MHC class II, CD40 and CCR2 by fluorescence activated cell sorter (FACS). In some samples, IFN-γ and 10 µg/ml immunoglobulin (Ig)G, sTNFR-Ig or IFN-γ and 10 µg/ml I-XPro or XPro1595 were added to wild-type cells (a). BMDMϕ from wild-type or TNFR1^−/− mice were also incubated with 100 ng/ml lipopolysaccharide (LPS) for 24 h prior to analysis of cell surface expression of MHC class II, CD40 and CCR2 by FACS. In some samples, LPS and 10 µg/ml IgG, sTNFR-Ig or LPS and 10 µg/ml I-XPro or XPro1595 were added to wild-type cells (b). Representative results from three independent experiments are shown.