Interferon gamma (IFNγ) ELISPOT assay
MultiScreen filter plates (EMD Millipore Corp. Billerica, MA) were coated with 10 µg/mL
mouse anti-porcine IFNγ monoclonal antibodies (Mabtech Inc. Cincinnati, OH), incubated
overnight at 4°C, washed three times with PBS and blocked with culture medium at 37°C for
1 h. PBMC (8 × 105
) were dispensed into each well and incubated with 10 µg/mL whole cell
sonicated Mhyo antigen (courtesy of M. Raymond, Iowa State University), 5 µg/mL