Anti cd25 mab

Icariin was purchased from MEYER company (Shanghai, China), whereas the antibodies used for western blotting were purchased from Cell Signaling Technology (Danvers, MA). The antibodies were anti-CD3 mAb (PE), anti-CD4 mAb (APC), anti-CD8a mAb (PE/Cy7), anti-CD25 mAb (APC/Cy7), anti-CD44 mAb (FITC), anti-CD62L mAb (APC/Cy7), anti-IFN-γ mAb (PE), HO-1 mAb (PE/Cy7), anti-CD16/32 mAb and anti-Foxp3 mAb (FITC) (eBioscience, San Jose, CA, USA).

BALB/c mice were randomly divided into four groups, including control group, vehicle group, Esculetin low-dose group (ES-L) and Esculetin high-dose group (ES-H). The back hair of all the mice was shaved in an area of 3 × 2 cm. To establish a mouse model of psoriasis, all groups except the control group were topically administered with a dose of 62.5 mg imiquimod cream (containing 5% imiquimod) on the back skin of mice for 7 consecutive days. Esculetin was prepared by 10% PEG400 solution. The mice of ES-L and ES-H groups were orally administered with esculetin at a dose of 50 and 100 mg/kg/day, respectively, for 7 consecutive days. While the control group and vehicle group were given 10% PEG400 solution daily for 7 consecutive days, all of the treatments began from the day when IMQ was administered. After 7 days, mice were sacrificed and their skin, spleens and lymph nodes were collected for further analyses. To deplete CD4⁺Foxp3⁺ Tregs, mice in some groups were treated i.p. with anti-CD25 mAb (Clone: PC61, eBioscience) at 0.2 mg on days 0, 2, 4, and 6 after IMQ was applied. This treatment with PC61 generally depletes around 80% of the CD4⁺CD25⁺ Tregs, as described in our previous studies (31 (link)).