Fig. 3: Histones induce a Th17 cell-specific response.
a Mean expression levels of T cell activation markers after 16 h co-culture with 0–10 µg/mL histone in the presence or absence (±) of αCD28 measured by flow cytometry n = 5. gMFI of each marker was normalized to the mean of no αCD28, 0 µg/mL condition and heat map created from the mean of the normalized values. b Differentiation of mouse naive T cells into Th1, Th17, and Treg cells in the presence of histones was measured by flow cytometric quantification of the expression of key factors IFNγ, IL-17 and FOXP3 from total live cells. c Graphical representation of b at various concentrations of histones with expression levels normalized to expression levels in the absence of histones; Th17 10 µg/mL histones n = 3, all others n = 4. d, e Frequency of cells expressing RORγt and f relative total RORγt expression at 32 h (calculated from the geometric mean) measured by flow cytometry n = 9. g, h Differentiation of mouse naive T cells into “pathogenic” Th17 cells with the addition of IL-23 in the presence of histones measured by flow cytometric quantification of IL-17 and GM-CSF after 5 days of culture n = 3. i Relative gene expression (fold-change) of histone-treated non-pathogenic (left) and pathogenic Th17 cells ± TGFβ after 3 days of differentiation compared to H2O-treated controls. Hprt expression was used as a housekeeping control n = 3. c, e, f, h Data bars show mean ± SEM. or i range and distribution, p > 0.05; “*”p < 0.05; “**”p < 0.01; “***”p < 0.001; “****”p < 0.0001. Where significance is not indicated, differences were ns. a, c Two-way ANOVA with Tukey’s multiple comparisons test, e, f, h paired student’s two-tailed t-test, i one-sample t-test (performed on ΔΔCt values). Data points represent individual mice. All data are representative of a minimum of two independent experiments. Source data are provided as a Source Data file.
