FIG 9.

T helper and memory T cell responses in pigs infected with nsp1β mutants. PBMCs collected at 7, 21, and 35 dpi were unstimulated or restimulated with the respective WT or mutant viruses that were used to infect pigs. Cells were immunostained using the pig-specific markers CD3, CD4, and CD8α, followed by intracellular IFN-γ detection. Each lymphocyte subset was grouped based on the combination of cellular markers: CD3⁺ CD4⁻ CD8α⁺ (CTLs/γδ T cells) (A), CD3⁺ CD4⁻ CD8α⁺ IFN-γ⁺ (activated CTLs/γδ T cells) (B), CD3⁺ CD4⁺ CD8α⁺ (T helper/memory cells) (C), CD3⁺ CD4⁺ CD8α⁺ IFN-γ⁺ (activated T helper/memory cells) (D), and CD3⁻ CD8α⁺ IFN-γ⁺ (activated NK cells) (E). A key explaining the treatment groups in panels A, B, D, and E is given in panel C. Each bar is the mean value ± SEM of data from 3 pigs. Statistical significance between wild-type and mutant virus-infected pig groups was determined by one-way ANOVA followed by Tukey's t test and is indicated with an asterisk (*, P < 0.05).