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. 2005 May;79(10):6052–6067. doi: 10.1128/JVI.79.10.6052-6067.2005

FIG. 10.

FIG. 10.

FIG. 10.

Activation of CD8+ lymphocytes from infected rabbits. (A) Increases in CD8+ IFN-γ+ cells upon stimulation of PBMCs isolated from rabbits at 3 (left) and 7 (right) days postinfection (dpi). PBMCs isolated from vMyx141KO-, vMyxgfp-, and mock-infected rabbits were cultured for 6 h in the presence of 25 ng/ml PMA plus 1 μM ionomycin and with Brefeldin A (BFA) for the final 4 h of the incubation. PBMCs were stained for the surface expression of CD8, fixed, permeabilized, and then stained for intracellular IFN-γ expression. Live gated events were assessed for anti-CD8-FITC and anti-IFN-γ-PE reactivity on a BD LSR II flow cytometer. Batch FACS analysis was then carried out with FlowJo software. (B) Representative dot plots of live gated PBMCs isolated from rabbits at 7 days postinfection from vMyxgfp- and vMyx141KO-infected rabbits. PBMCs were cultured for 6 h in the presence of 25 ng/ml PMA plus 1 μM ionomycin and with BFA for the final 4 h of incubation. PBMCs were stained for the surface expression of CD8, fixed, permeabilized, and stained for intracellular IFN-γ expression. Quadrant numbers represent percent total events assessed for anti-CD8-FITC and anti-IFN-γ-PE reactivity on a BD LSR II flow cytometer. Batch FACS analysis was then carried out with FlowJo software.