Pharmacological inhibition of RORγt suppresses the Th17 pathway and alleviates arthritis in vivo
Fig 2
Cpd 1 is a potent and selective RORγt inhibitor in in vitro cellular assays.
(A) Overexpression of RORγt results in IL-17A production. HUT78 T-cells, stably transduced with human full-length RORγt or control vector, were stimulated with anti-CD3 antibody plus PMA for 48 hrs and IL-17A production was quantified by ELISA. (B) HUT78 T-cells expressing RORγt were incubated with serial dilutions of cpd 1 at the beginning of the stimulation with PMA and anti-CD3 antibody and after 48 hrs IL-17A concentrations were measured by ELISA. Representative example of a concentration-response curve from five independent experiments with triplicate readings is shown. Activity of cpd 1 against the nuclear hormone receptors RORα (C) or RORβ (D). Jurkat T-cells stably expressing GAL4 upstream activator sequences that control the transcription of the luciferase reporter gene were transfected with a GAL4-DNA binding domain fusion construct containing the RORα or RORβ-LBD. Cells were incubated for 24 hrs with cpd 1, followed by cell lysis and measurement of luciferase activity. Graphs are representative of three independent experiments performed in duplicates.