Figure 2. VV-specific immune responses in FoxP3^KO mice.

Splenocytes of cutaneously VV-infected FoxP3^KO and WT control mice were analyzed eight days p.i.. Data were compiled of three independent experiments. (A) Splenocytes were stained for CD8, VV-specific TCR (using B8R_20–27 MHC class I pentamers) and CD45R/B220 and analyzed by flow cytometry. Representative dot plots are shown (VV infected FoxP3^KO mice n = 17, WT n = 8). Uninfected mice did not have any VV-specific CD8⁺ T cells (Figure S3). (B) Splenocytes were stimulated in vitro with the VV peptide, B8R_20–27. Intracellular IFN-γ as well as CD8 and CD4 surface staining were performed and cells were analyzed by flow cytometry gating on CD8⁺ positive cells. Representative dot plots are shown (n = 7). (C) Cytokine levels in supernatants of splenocytes from VV-infected FoxP3^KO and WT control mice stimulated with irradiated VV-infected (black columns) or uninfected (white columns) splenocytes from naïve WT mice. Cytokines were determined by ELISA 24 hours (IFN-γ), 48 hours (IL-4) and 72 hours (IL-2) after stimulation (VV-infected FoxP3^KO mice n = 9, all others n = 7). ** p < 0.005, ***p < 0.0005