Fig. 2.

Neither anti-HMGB1 antibodies nor box A suppressed LPS activity in the Limulus assay; m2G7 did not suppress pro-inflammatory cytokine release induced by TNF, peptidoglycan (PEG), Poly I: C or CpG DNA from human macrophages. a. Polyclonal anti-HMGB1 antibody (pAB, 1 µg/ml), m2G7 (1 μg/ml) or box A (10 μg/ml) were added to LPS (from 0 to 2 U/ml) solution and incubated at 37 °C for 30 min and limulus assay was performed. Results are representative of 2 separate measurements each done in triplicate. b. Primary human macrophages in 96-well culture plate (5 × 10⁴ cells/well) were stimulated with mouse TNF in the presence or absence of m2G7 for 16 h. IL-1β released was measured. N = 3 experiments. c. Primary human macrophages in 96-well culture plate (5 × 10⁴ cells/well) were stimulated with TLR 2 agonist PGN (5 μg/ml), TLR3 agonist Poly I:C (50 μg/ml) or TLR9 agonist CpG DNA (1 μM) for 16 h. TNF released was measured. N = 3 separate experiments