Fig. 4

Exposure to C. concisus Zot enhances the response of THP-1 macrophage-like cells to E. coli K12. The production of pro-inflammatory cytokines and phagocytosis by THP-1 macrophage-like cells enhances the response to E. coli K12 following exposure to C. concisus Zot were assessed. THP-1 cells were treated with EP and EP-ZotP14UCO-S1 at 50 μg/100 μl for 2 h. The proteins were removed and cells were washed with DPBS. Measurement of cytokines levels (a and b) the cells were then incubated with E.coli K12 Bioparticles for an additional 2 h and the supernatants were used to measure TNF-α and IL-8 using commercially available ELISA kits. Phagocytosis (c) the phagocytosed E. coli K12 Bioparticles were measured using a Fluorescence Plate Reader. The values in the negative control were subtracted from the positive control and experimental wells. The levels of phagocytosis in THP-1 macrophage-like cells treated with different proteins were expressed as percentage relative to that of the positive control. Data are shown as the mean ± SD from triplicates and are representative of two independent experiments. ** Indicates P < 0.01. EP-ZotP14UCO-S1 constituted of E. coli proteins and the Zot of C. concisus strain P14UCO-S1 expressed in E. coli. EP control E. coli proteins