Figure 7.

Vaccination with MAYV/IRES strain induces a better splenic immune response after wt MAYV challenge. Flow cytometric analysis was conducted to evaluate the impact of vaccination on the cellular immune response to wt MAYV infection of mice previously vaccinated with wt MAYV or MAYV/IRES. Six-week-old BALB/c mice were inoculated with 2 × 10⁵ PFU/50 μL of wt MAYV or MAYV/IRES via intraplantar (i.pl.) route and 28 days later mice were challenged with 2 × 10⁵ PFU/50 μL of wt MAYV at the same hind paw. After 3 days, animals were culled, and splenocytes were isolated for flow cytometry. Cells were incubated in the presence of the protein transport inhibitor brefeldin A, followed by staining with antibodies against surface markers expressed by DCs (a–d), monocytes/macrophages (e–g), T CD8⁺ (e–i) and T CD4⁺ (h–l) lymphocytes. Splenocytes were then fixed, permeabilized and incubated with antibodies against intracellular TNF-α (c,g), IL-17 (k) and Foxp3 (l). Gating strategy is illustrated in Supplemental Fig. 4. Data is expressed as mean ± SEM of at least 5 animals. MOCK- not-infected. *p < 0.05 when compared to control uninfected cells (MOCK). ^#for p < 0.05 when compared to wt MAYV vaccinated mice, as assessed by one-way ANOVA followed by Newman-Keuls post-test.