Expression of PPRV glycoproteins and functionally active ovine IL-2 and GMCSF, in vitro. (A) HEK 293 cells were infected with Ad-F, Ad-H or Ad-GMCSF, and Vero-SLAM cells were infected with PPRV or mock infected. Total cell lysates were analysed by Western blot using an anti-PPRV F monoclonal antibody or polyclonal rabbit anti-PPRV H serum as indicated. (B, C) HEK 293 cells were infected with Ad-IL-2, Ad-GMCSF or Ad-H, or mock infected overnight and cell lysates (CLs) and supernatants (SNs) were harvested. The biological activity of GMCSF in CLs and SNs combined from Ad-GMCSF, Ad-H or mock-infected cells was determined by analysis of proliferation of goat bone marrow cells. The biological activity of IL-2 in SNs from Ad-IL-2, Ad-H or mock-infected cells was determined by analysis of proliferation of goat blood lymphocytes. Proliferation was measured by 3H-thymidine incorporation after 7 days incubation and expressed as the mean stimulation index ± S.D. of triplicate wells.