Figure 6.

Analysis of CD4⁺ and CD8⁺ T cells that produce IFN-γ following infection with TMEV. (A) Mononuclear cells from the CNS of mice at 7 and 21 dpi were cultured for 6 h in the presence of PBS or a mixture of 1 μM of viral peptides and assessed the production of intracellular IFN-γ using flow cytometry. The data are representative of three independent experiments. (B) Splenocytes from mice at 7 and 21 dpi were cultured for 72 h in the presence of PBS, a mixture of 1 μM viral peptides or anti-CD3/CD28 antibodies. The production of IFN-γ in the culture supernatants were assessed using ELISA. Mixtures of CD4⁺ T cell-specific epitopes (SP mix; VP1_233–250, VP2_74–86, and VP3_24–37; NSP mix, 3D_6–23 and 3D_20–38) and CD8 ⁺ T cell-specific epitopes (CD8 mix; VP3_159–166, VP3_173–181, and VP1_11–20) were used. Mixtures of plate-bound anti-CD3 and anti-CD28 antibodies (1 μg each) were used to stimulate pan-T cells. Values given are the means (± SD) of the results from triplicate wells. ^*p < 0.05; ^**p < 0.01; ^***p < 0.001.