MOESM7 of Frequency of Th17 cells correlates with the presence of lung lesions in pigs chronically infected with Actinobacillus pleuropneumoniae

Additional file 7. Co-production of TNF-α and IL-17A by CD4 + T cells in peripheral blood. Phenotyping and intracellular cytokine staining were performed on PBMC following overnight in vitro stimulation (APP-CCE, medium, PMA/Ionomycin). (A) Living CD4+ T cells were gated (not shown; see Additional file 1) and further analyzed for production of TNF-α and IL-17A. Data from the same animals as in Additional file 2 are shown. Approximately 3 × 105 (APP and medium) and 5 × 104 (PMA/Ionomycin) cells are shown in the contour plots. (B) Frequency of IL-17A/TNF-α co-producing CD4+ T cells in PBMC of infected animals (red dots) and control animals (blue dots) during acute and chronic phase. Numbers next to colored dots indicate numbers of individual animals. Median percent values are indicated by black bars. Medium-corrected percentage values are presented (% of IL-17A+ TNF-α+ cells within total CD4+ T cells for APP-CCE stimulation minus % of IL-17A+ TNF-α+ cells within total CD4+ T cells for medium incubation). Arrow heads are introduced in the main text