Fig. 3

SJMHE1 increased CD4⁺CD25⁺ Foxp3⁺ T-cells in DTH mice. BALB/c mice were sensitised with OVA alone or combined with 30 μg of SJMHE1. Challenge with OVA occurred 7 days later; 24 h after the challenge, spleen and LNs from each mouse were pooled. Single-cell suspensions were prepared, and red blood cells were lysed. a Flow cytometry for CD3, CD4, CD25, and Foxp3 was performed, and data are expressed as the mean ± SD of 18 mice from three independent experiments; b Flow cytometry for CD3, CD4, CD25, and CTLA4 was performed. Data are expressed as the mean ± SD of 18 mice from three independent experiments; c Analysis of CD4⁺CD25⁺Foxp3⁺ T-cells from pooled splenic and LN cells by flow cytometry. Data are representative of the experiments; d Analysis of CD4⁺CD25⁺CTLA4⁺ T-cells from pooled spleen and LN cells by flow cytometry. Data are representative of the experiments. Values indicate the percentage of events in the indicated quadrant. Asterisks indicate significant differences analysed using the independent Student’s t-test (***P < 0.001)