Antibody Inhibition of a Viral Type 1 Interferon Decoy Receptor Cures a Viral Disease by Restoring Interferon Signaling in the Liver
Figure 5
Identification of a mAb that inhibits the biological activity of ECTV T1-IFNbp.
A) ELISA plates were covered with recombinant T1-IFNbp and the presence of the protein was detected with the indicated mAbs or isotype control (IC). Data are representatives of 2 independent experiments with similar results. B) L929 cells (105) were incubated with 10 ng recombinant T1-IFNbp at 37°C for 1 h, thoroughly washed, and the binding of mAb 10G7 (blue line) or 10F3 (green) to the cell surface was analyzed by flow cytometry. C) L929 cells were infected with ECTV-GFP (upper panels) or ECTV-Δ166-GFP (lower panels) at MOI of 0.05 for 16 h. GFP expression and the binding of IC, 10G7 or 10F3 mAbs were determined by flow cytometry as indicated. D) 10 ng of recombinant ECTV T1-IFNbp was incubated with 10 µl rabbit anti-IFNbp or 10 ng of indicated mAbs for 30 min at 37°C, incubated with 1U mIFN-α for another 30 min at 37°C. The cocktail was then added to L929 cells in 96-well plates. The cells were incubated with this cocktail for 24 h, infected with VSV-eGFP at MOI of 0.1 for 16 h and observed for GFP fluorescence under the microscope.