Sars cov 2 s1 mixed peptide pools

Murine IFN-γ, IL-2, and IL-4 ELISPOT were performed following the kit instructions (Mabtech). In brief, splenocytes harvested from immunized mice were grinded into a single cell suspension. The suspended cells were seeded in 96-well ELISPOT plates at density of 250,000 cells/well in duplicates. Cells were stimulated with 0.4 μg/well SARS-CoV-2 S1 mixed peptide pools (Mabtech) at 37°C for 18–20 hours before counting the spot numbers by Mabtech ASTOR ELISpot reader.

The splenocytes harvested from immunized mice were grinded into a single cell suspension. The suspended cells were seeded on 96-well U-bottom plates at density of 2 × 10⁶ cells/well and stimulated with SARS-CoV-2 S1 mixed peptide pools (2 μg/mL, Mabtech) for 18–20 hours. The resulting cells were harvested for surface and intracellular markers staining. Cells were first incubated with anti-CD4 and anti-CD8 antibodies for surface staining followed by intracellular staining with anti-IFN-γ, anti-IL-4, and anti-Granzyme B antibodies (Biolegend). All flow cytometry data was acquired by Navious EX flow cytometer (Beckman Coulter) and analyzed by Kaluza software.