Induction of Release of Tumor Necrosis Factor and IL-6from Human Mononuclear Cells byBacteroidesstrains

Abstract The role of anaerobic Gram-negative bacteria in inducing cytokines during mixed infections involving aerobic and anaerobic bacteria is relatively poorly defined. The purpose of this study was to establish whether or not intact Bacteroides fragilis and related species, isolated from severe infections and from the faeces of healthy persons are capable of releasing tumor necrosis factor (TNF) and IL-6 from human mononuclear cells and whole blood. The purified lipopolysaccharides of Bacteroides fragilis strain (No. 7), extracted by the aqueous phenol method from BHI cultures and from BHI culture supplemented with 5% horse serum, were also tested. TNF release was detected by the WEHI 164-dependent bioassay and IL-6 production by the B-9 cell-dependent bioassay. Heat-inactivated Bacteroides strains belonging to different species were able to induce TNF (1×10 1 −5×10 2 U/mL) and IL-6 (1×10 1 −5×10 5 pg/mL) release from human mononuclear cells. When whole blood was used, the production of TNF and IL-6 was more pronounced (very probably because of the presence of certain serum factors). The culturing conditions (the presence of 5% horse serum in the BHI broth) influenced the inducing activity of almost all strains tested. The isolated lipopolysaccharide of Bacteroides fragilis strain No. 7 proved to have a rough profile on PAGE. There were no differences in TNF and IL-6 induction when the lipopolysaccharides of the strain was cultured in BHI or in BHI supplemented with 5% horse serum. Bacteroides strains often outnumber Enterobacteriaceae in the faeces and in mixed infections, and their role in inducing and/or modulating the host response in septic shock should not be overlooked.