Y Baine, B M Stankunas, P Miller, C Hobbs, L Tiberio, J Koch, K Yoon, D Sawutz, C Surowy
Department of Immunology, Sanofi Winthrop Pharmaceuticals Research Division, Collegeville, PA 19426, USA.
Journal of immunology (Baltimore, Md. : 1950) 1995 Apr 15IL-2-mediated T cell proliferation is a critical early event in the inflammatory process. Formation of the NFAT-1 transcriptional complex on the IL-2 promoter is essential for IL-2 transcription. Using a cell line that is stably transfected with a trimer of the NFAT-1 regulatory element linked to a lac-Z reporter gene, we screened for inhibitors of NFAT-1-mediated beta-galactosidase activity. WIN 61058 and WIN 53071 were identified as microM inhibitors. These compounds also inhibited beta-galactosidase mRNA levels. Similar inhibition of NFAT-1-mediated gene expression was observed in a second cell line, which is stably transfected with NFAT-1 regulatory elements linked to the reporter gene for sCD8. At 10 microM, both compounds inhibited IL-2 mRNA and protein levels in the NFAT-1-linked lac-Z transfectants, and in human lymphocytes. Both compounds inhibited the mixed lymphocyte reaction, and this inhibition was reversed by exogenous IL-2. WIN 53071 inhibited IL-2 production induced in the calcium-dependent PMA and ionomycin pathway. Conversely, calcium-independent anti-CD28 Ab and PMA-induced IL-2 production was resistant. Both compounds altered the NFAT-1 transcriptional complex, causing its retarded mobility on gels. By these functional criteria, we believe we have identified two structurally distinct, novel inhibitors of NFAT-1-mediated transcription.
Y Baine, B M Stankunas, P Miller, C Hobbs, L Tiberio, J Koch, K Yoon, D Sawutz, C Surowy. Functional characterization of novel IL-2 transcriptional inhibitors. Journal of immunology (Baltimore, Md. : 1950). 1995 Apr 15;154(8):3667-77
PMID: 7706710
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