Am80 inhibits donor Th1 and Th17 cells in vitro and in vivo. Sublethally irradiated BALB/c recipients were transplanted from WT B10.D2 donors. (A-B) PLN cells from recipients (n = 3-6 per group) on day 14 were treated with Am80 or vehicle solution for 24 hours, the supernatants were collected, and ELISA was performed to determine the cytokine levels. Graphs represent the levels of cytokines secreted per 1 × 10⁶ whole stimulated PLN cells. The data are from 1 representative of ≥ 3 independent experiments. (C-D) After BMT, recipients (n = 4-6 per group) were administered oral Am80 (1.0 mg/kg of body weight) or vehicle solution daily from day 0. PLNs of the recipients were stained for intracellular IFN-γ and IL-17. (C) The percentage and absolute number of IFN-γ⁺ and IL-17⁺–producing CD4⁺ T cells. Data are from 1 representative of ≥ 2 repeated experiments. (D) PLN cells from recipients (n = 3-6 per group) treated with Am80 or vehicle on day 16 were stimulated with PMA and ionomycin. Five hours later, the supernatants were collected to determine cytokine levels by CBA. Graphs represent the levels of cytokines secreted per 1 × 10⁶ whole stimulated PLN cells. The data are from 1 representative of ≥ 3 independent experiments. The means (± SEs) of each group are shown; *P < .05 and **P < .01.