Cytokine Secretion in Mouse Spleen Cells

The levels of the cytokines IL-2 and IFN-γ secreted by mouse spleen cells were detected by ELISA with the double-antibody sandwich method. The capture antibody was diluted with PBS to the appropriate concentration (3 μg/mL for IL-2 and 4 μg/mL for IFN-γ). Prepared spleen cells were added to 96-well plates (1 × 10⁶ cells per well), and gE protein was added to the wells other than the negative and positive control wells as a stimulus. PMA + ionomycin (10 μg/mL, DAKEWE, Beijing, China) was added to the 96-well plates (Corning Inc., Corning, NY, USA) as a positive control. The cells were incubated overnight at 37 °C and 5% CO₂, and the cell supernatants were collected to detect the levels of IL-2 and IFN-γ [22 (link)].