Gs 9620

Neutralizing antibodies against mouse IFN- alpha receptor or IFN- gamma (Bio X Cell) were i.p. injected at a dose of 150 μg/mouse into 1-week-old wt-129 mice, either before or after i.p. inoculation with EV-A71 clinical isolate F23 (10⁸ PFU/mouse). The neutralizing antibody against the IFN-alpha receptor was injected 1 day before EV-A71 inoculation, followed by repetitive antibody treatments on 1 and 3 dpi. Recombinant mouse IL28A and IL28B were both purchased from R & D Systems®. Various doses of mouse IL28 was i.p. injected into mice at dpi = 1 (Additional file 4: Figure S4a). Recombinant mouse IFNαA (PBL Assay Science Co.), GS9620 (Cayman Chemical) and R837 (InvivoGen), were orally or i.p. injected into wt-129 mice infected with EV-A71, at the dose of 300 IU/g (mouse IFNαA), 3.0 μg/g (GS-9620), 4.5μg/g (GS-9620), and 5.0 μg/g (R837), respectively.

R837 (selective TLR7 agonist), R848 (TLR7 & TLR8 agonist) and TL8-052 (selective TLR8 agonist) were acquired from Invivogen and stored at −20°C. TLR-agonist R837 was used at a concentration of 5 μg/mL. R848 at 10 μg/mL and TL8-052 at 5 μg/mL.
Vesatolimod, Gardiquimod, Telratolimod, Selgantolimod and Motolimod were acquired from a commercial distributor (MedChemExpress) and stored as per the manufacturer’s instructions. Concentrations were used in increasing concentration to establish working concentrations as follows: TLR-agonist GS-9620/Vesatolimod, 0.1 μM, 1 μM, 3 μM, 10 μM, 20 μM, 30 μM, 60 μM (selective TLR7 agonist), Gardiquimod, 0.1 μM, 1 μM, 3 μM, 10 μM, 20 μM, 30 μM (selective TLR7 agonist), 3 M-052/Telratolimod, 0.1 μM, 1 μM, 3 μM, 10 μM, 20 μM, 30 μM (TLR7 & TLR8 agonist), GS-9688/Selgantolimod, 0.1 μM, 1 μM, 3 μM, 10 μM, 20 μM, 30 μM (selective TLR8 agonist), MTX-1337/Motolimod, 0.1 μM, 1 μM, 3 μM, 10 μM, 20 μM, 30 μM (selective TLR8 agonist).
PolyIC-lyovec (LMW) was acquired from Invivogen and dissolved in LAL-water as per the manufacturer’s instructions and used at a concentration of 2 μg/mL. Salmonella Lipopolysaccharide (LPS) was used as positive controls at a concentration of 10 ng/mL.