Screening assay for the identification ov novel immunosuppressives using cultured T cells
AssigneeThe Regents of the University of California
InventorsArthur Weiss, James Fraser
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Abstract
A method for identifying compounds capable of inducing immunosuppression by inhibiting the CD28 signal transduction pathway and T cell comprises exposing cultured T cells to one or more test compounds. The T cells are obtained from a T cell line which stably incorporates DNA sequence comprising in reading frame an enhancer region responsive to a CD28-regulated nuclear binding protein and a marker gene. The cells are cultured under conditions which result in activation of both the T cell receptor and the CD28 receptor, resulting in enhanced expression of the marker gene. Test compounds which inhibit expression of the marker gene are considered as candidates for immunosuppressive drugs.
Claims4
A method for screening test compounds for the ability to inhibit the CD28 signal transduction pathway in T cells, said method comprising:
- exposing TCR-activated cultured T cells to conditions selected to induce the CD28 signal transduction pathway, wherein a DNA sequence comprising an enhancer region responsive to a CD28-regulated nuclear binding protein, a promoter, and a marker gene is stably incorporated in said T cells, and wherein said T cells are Jurkat-IL2-Luc cells, ATCC Accession No. CRL 11509;
- administering test compounds to the cultured cells;
- and identifying which test compounds suppress expression of the marker gene.
Dependencies are inferred from the claim text.