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Enzyme-linked immunospot (ELISPOT) assay
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Splenocytes and lung lymphocytes were stimulated with 500 ng/well IIV or 5 μg/well HA-specific T cell epitope peptide mixture (LYEKVRNQL, GIPSRISI, GMVDGWYG, VRPVTSGCF, RGFFGAIAGF, and IYSTVASSL) for 48 h at 37 °C. HA peptides were synthesized by Peptron (Daejeon, Korea). The ELISPOT assays for the detection of IFN-γ or IL-4-secreting T cells were conducted using mouse IFN-γ and IL-4 ELISpotBASIC (Mabtech, Stockholm, Sweden). Each step was performed according to the manufacturer’s instructions.
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