Figure 1.

Effect of Ara-LAM pretreatment on the frequency and phenotype of T regulatory cells. (A) CD4⁺ T cells (1 × 10⁶) were purified by MACS from spleen of infected and indicated treatment groups of mice 28 days after infection, plated aseptically followed by fixation and staining for PE-conjugated CD25 and FITC-conjugated Foxp3 as mentioned in the section ‘Materials and Methods’ and analyzed by flow cytometry. Data are from one of three representative experiments. (B) In a separate experiment, CD4⁺CD25⁺ Treg cells (2 × 10⁵) were purified from spleen of differently treated mice by MACS as described in the section ‘Materials and Methods’ and collected in TRIZOL for mRNA extraction. The expression of Foxp3, GITR and CTLA4 mRNAs was analyzed by semiquantitative RT-PCR. The data are representative of three independent experiments, which yielded similar results.