Effect of Tac in Th17 subpopulations of CD4+T lymphocytes from the peripheral blood of early-post transplant recipients.

We used flow cytometry to examine how Tacrolimus regulates in vitro Th17 subpopulations of CD4+ T lymphocytes in Th17-polarizing condition. PBMC from renal transplant recipients were preincubated for 1 h in the presence of Tacrolimus and stimulated with Th17-polarizing condition for 48 h. Anti-CD3 (1 µg/ml), anti-CD28 (1 µg/ml), IL-1b (20 ng/ml), IL-6 (20 ng/ml) and IL-23 (20 ng/ml) were added to stimulate the differentiation of Th17 cells. Neutralizing antibodies to IFN-gamma (2 µg/ml) and IL-4 (2 µg/ml) were added in some experiments (R&D Systems). Flow cytometry of intracellular Th17 in CD4+ T cells stimulated in the presence of Th17-polarizing condition, assessed after 48 h and then stimulated for 4 h with PMA and ionomycin in the presence of GolgiStop. The data are representative of three independent experiments. The values are expressed as the mean ± SEM.