Viral Vectored Blood Stage Malaria Vaccine Figure 8. T cell responses induced in mice by vaccination with the mono-allelic and bi-allelic constructs. 5–6 week old female BALB/c mice (n = 6) were immunized as in Figure 6. Splenocytes were isolated two weeks after the final immunization (day 70) and stimulated with four peptide pools (Common, 3D7, FVO and C-terminus) or no peptide (Unstim) for 5 hours. After this time the cells were surface stained for CD4 and CD8 and then intracellularly stained for IFN-c, TNFa and IL-2 and analysed by flow cytometry for % of responding cells per total CD4+ or CD8+ T cell subset. % CD4+ and CD8+ T cells positive for IFN-c, TNF-a or IL-2 following the different vaccination regime is shown (8A-H). Individual data points and the median are shown. The asterisk indicates a significant difference between the groups by ANOVA (* P #0.05, ** P #0.01). doi:10.1371/journal.pone.0020977.g008 PLoS ONE | www.plosone.org 11 June 2011 | Volume 6 | Issue 6 | e20977