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. 2021 Feb 3;11:603059. doi: 10.3389/fimmu.2020.603059

Figure 5.

Figure 5

Development of tolerance to new oral antigens is abrogated following depletion of peri-weaning Tregs. (A) Experimental schematic describing labeling of Tregs with DTR using tamoxifen followed by deletion of labeled cells on DOL 56, and DTH response (left, B and C) or transfer of OTII cells (right, D) FoxP3iDTR mice were injected with tamoxifen on DOL14 (light blue) or DOL24 (purple). C57Bl6 mice were used as controls and injected with a vehicle (black) or tamoxifen on DOL24 (gray). All groups were injected with diphtheria toxin on DOL56 for deletion of labeled cells on DOL 56. (B) Increase in footpad thickness following 7 days of Ova in drinking water, immunization, and footpad challenge with Ova, analyzed by one-way ANOVA with Dunnett’s test for multiple comparison. (C) Change in temperature following footpad challenge, analyzed by two-way ANOVA with Tukey’s test for multiple comparisons. (D) Number of Ova-specific OTII cells in the colon LP 7 days following adoptive transfer and dietary Ova, analyzed by one-way ANOVA with Dunnett’s test for multiple comparison. n = 6 mice per group in panels (B, C) from two independent experiments, c displays data from one representative experiment, n = 3 mice in panel (D) * denotes statistical significance < 0.05.