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. 2020 Sep 4;11:2071. doi: 10.3389/fimmu.2020.02071

FIGURE 4.

FIGURE 4

In vitro evidence suggests limited anergy of CD4+ cells in dTg HEL/TCR mice. CD4+ T cells isolated from 3A9 TCR or dTg HEL/TCR mice aged P50-P70 (in the presence of mitomycin C-treated antigen-presenting feeder cells from B10. Br WT mice) were incubated with decreasing concentrations of HEL protein (10– 5–10– 11 M over 72 h), and their proliferation assessed using flow cytometry to measure CFSE dilution. Limited anergy of dTg HEL/TCR CD4+ T cells (i.e., reduced specific responsiveness to HEL reflected in consistently low proliferation) was observed at and below a HEL antigen concentration of 10– 10 M. Significant differences were assessed using one-way ANOVA, followed by Tukey post hoc test with ***p < 0.001, ****p < 0.0001 on a 95% level of confidence. The experiment was independently repeated twice.