Isolation of DCs and T cells from Splenic Lymphocytes

Splenic lymphocytes were collected and cultured as described previously55 (link). Part of splenic lymphocytes was incubated with anti-CD11c microbeads (Miltenyi Biotec, Bergisch Gladbach, Germany) and separated using AutoMacs (Miltenyi Biotec). Selected cells were considered CD11c⁺ DCs. Negatively selected cells were stained with anti-CD3 FITC (Miltenyi Biotec), and subsequently incubated with anti-FITC microbeads (Miltenyi Biotec) to obtain CD3⁺ T cells. DCs and CD3⁺ T cells were cultured in RPMI-1640 medium supplemented with 10% FBS for further detection of the percentages of PRRSV-specific CD3⁺ T cells producing IFN-γ and IL-4.