Briefly, antibody-coated membrane plates were seeded with5105 cells per well and stimulated with 0.1 ml of 5
106 TCID50/ml live TX98, CO99 virus, or uninfected MDCK culture supernatant intriplicate. Followingan 18–20 h incubation period, plates were developed according to manufacturer's recommendations. Dryplates were scanned and analyzed withS5UVCTL-ImmunoSpot in strumentation and software (CellularTechnology Ltd., ShakerHeights,OH). Individual pig responses were calculated as the average number of spots per triplicate well within a stimulation treatment.