Frequencies of T cells releasing cytokines in response to Avi-OVA coated BCG-immunization.

<p>Splenocytes from immunized mice with PBS, BCG Wild-type, BCG WT surface decorated with Avi-OVA, BCG-p19-OVA and BCG-p19-AviOVA were stimulated with recombinant OVA protein for 16 hours followed by a 5 h-period treatment with Brefeldin A. Cells were then washed and stained first with PE-Cy7 anti-CD4 (A) or PE anti- CD8 Ab (B) then AF647 anti-IFN-γ Ab. Cells were then washed and analyzed by FACS. Results are expressed as two-parameter dot plots to show the average frequencies± SEM of IFN-γ producing cell subsets in CD4⁺ and CD8⁺ populations from two animals/group. Data shown are representative of three independent experiments. The data in graphs (C) are expressed as the mean of absolute number± SEM of IFN-γ releasing CD4⁺ T cells (left graph) or IFN-γ releasing CD8⁺ T cells (right graph) from two animals/group and three independent experiments. *P<0.05; **P<0.01; ***P<0.001.