qPCR and ELISPOT.rar

FMDV has been shown to efficiently replicate in the mouse model, with peak viral loads detected in the spleens on the 5 day post-infection (dpi)

CD8⁺ and CD4⁺ T cells were isolated from splenocytes using magnetic bead-based positive selection kit (Miltenyi Biotec, Germany). A total of 2 × 10⁵ purified CD8⁺ or CD4⁺ T cells were co-cultured with 10⁵ DC2.4 cells, serving as antigen-presenting cells (APCs), in a 96-well Mouse IFN-γ precoated ELISPOT kit (Dakewe, 2210003, China). Cells were stimulated in triplicate with the specified peptide (10 µg/mL) following the manufacturer’s instructions. Positive (PMA-Ionomycin) and negative (DMSO) controls were included in all experiments to ensure assay validity.