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. 2017 Oct 24;8:1340. doi: 10.3389/fimmu.2017.01340

Figure 5.

Figure 5

DNA vaccination against spring viremia of carp virus (SVCV) induces the formation of virus-specific T cells. PBLs were isolated 3 months after vaccination from non-vaccinated (pcDNA3) and vaccinated (pcDNA3-SVCV-G) carp. Carboxyfluorescein succinimidyl ester (CFSE)-labeled PBLs were stimulated for 6 days at 27°C with recombinant Ifnγ2 (100 ng/mL) or Il10b (0.25 U/mL) alone, or in combination with SVCV (MOI of 25). As a control, conditioned medium was used. T-cells were identified using a cross-reactive anti-Zap70 antibody, and proliferation was analyzed by flow cytometry. (A) Percentages of proliferating Zap70+ T cells are expressed relative to the respective conditioned medium control after subtraction of the percentage of proliferating cells in the medium control. For each group, bars represent average and SD of n = 7 fish/stimulus, except for the Il10b stimulated samples for which n = 3 fish were used. Asterisk (*) indicates a significant differences between the pcDNA3 and pcDNA3-SVCV-G group as assessed by one-way ANOVA. (B) Representative histogram plots of CFSE-labeled Zap70+ T cells from non-vaccinated (pcDNA3, left panel) and vaccinated (pcDNA3-SVCV-G, right panel) carp of the experiment in panel (A).