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. 2019 Jul 16;10:1638. doi: 10.3389/fimmu.2019.01638

Figure 6.

Figure 6

CD4+ T cell differentiation upon VISTA engagement. CD4+ T cells were negatively isolated from PBMCs. Cells were cultured in 96-well flat-bottom plates and stimulated for 5 days at 37°C in anti-CD3 (2.5 ug/mL) coated plates simultaneously coated with either control Ig or VISTA-Ig (both 10 ug/mL). Soluble anti-CD28 was added at 250 ng/mL. After 5 days, CD4+ T cell differentiation was assessed by measuring the expression of intracellular transcription factors (T-bet, RORɤt, BCL6, GATA3, and FoxP3). (A) Frequency of transcription factor expression of CD4+ T cells from healthy controls when engaged to control Ig (light blue) and VISTA-Ig (dark blue) (n = 6). (B) Frequency of transcription factor expression of CD4+ T cells from GCA patients when engaged to control Ig (light orange) and VISTA-Ig (dark orange) (n = 6). (C) Radar plot of the frequency of transcription factor expression of CD4+ T cell from HC (blue) and GCA patients (orange) combined. Individual values represent the median. Significant differences by Wilcoxon signed-rank test are indicated: *P < 0.05, ns, non-significant.