Immunosuppressant PG490 (triptolide) inhibits T-cell interleukin-2 expression at the level of purine-box/nuclear factor of activated T-cells and NF-kappa B transcriptional activation

PG490 (triptolide) is a diterpene triepoxide with potent immunosuppressive and antiinflammatory properties. PG490 inhibits interleukin(IL)-2 expressio n by normal human peripheral blood lymphocytes stimulated with phorbol 12-m yristate 13-acetate (PMA) and antibody to CD3 (IC50 of 10 ng/ml), and with PMA and ionomycin (Iono, IC50 of 40 ng/ml). In Jurkat T-cells, PG490 inhibi ts PMA/Iono-stimulated IL-2 transcription. PG490 inhibits the induction of DNA binding activity at the purine-box/antigen receptor response element (A RRE)/nuclear factor of activated T-cells (NF-AT) target sequence but not at the NF-kappa B site. PG490 can completely inhibit transcriptional activati on at the purine-box/ARRE/NF-AT and NF-kappa B target DNA sequences trigger ed by all stimuli examined (PMA, PMA/Iono, tumor necrosis factor-alpha). PG 490 also inhibits PMA-stimulated activation of a chimeric transcription fac tor in which the C-terminal TA1 transactivation domain of NF-kappa B p65 is fused to the DNA binding domain of GAL4. In 16HBE human bronchial epitheli al cells, IL-8 expression is regulated predominantly by NF-kappa B, and PG4 90 but not cyclosporin A can completely inhibit expression of IL-8. The mec hanism of PG490 inhibition of cytokine gene expression differs from cyclosp orin A and involves nuclear inhibition of transcriptional activation of NF- kappa B and the purine-box regulator operating at the ARRE/NF-AT site at a step after specific DNA binding.