A HUMAN IL-1 INHIBITOR FOR THE TREATMENT OF SEPTIC SHOCK

WE PROPOSE TO PURIFY AN IL-1 INHIBITOR FROM THE SUPERNATANT OF MONOCYTES CULTURED ON PLATES COATED WITH IGG. THIS INHIBITORY ACTIVITY HAS BEEN SHOWN TO BE ABOUT 22 KILODALTONS IN MOLECULAR WEIGHT AND TO INHIBIT IL-1 IN SEVERAL ASSAYS, NAMELY IL-1 MEDIATED THYMOCYTE PROLIFERATION, COLLAGENASE RELEASE BY ARTICULAR CHONDROCYTES, AND PGE(2) RELEASE BY BOTH SYNOVIAL AND DERMAL FIBROBLASTS. WE WILL PRODUCE LARGE QUANTITIES OF THE INHIBITOR-CONTAINING SUPERNATANT AND PURIFY THE PROTEIN BY CONVENTIONAL CHROMATOGRAPHIC TECHNIQUES. INITIALLY WE WILL USE THE THYMOCYTE-PROFILERATION ASSAY TO MONITOR PURIFICATION BUT WILL IMMEDIATELY BEING DEVELOPING A FASTER PGE(2)-RELEASE ASSAY TO SPEED UP PURIFICATION. WE ALSO WILL BEGIN DEVELOPING MONOCLONAL ANTIBODIES TO THIS MOLECULE. OUR PHASE-I GOALS ARE TO DEVELOP A RAPID ASSAY FOR THIS INHIBITOR AND TO ATTAIN A 400-FOLD PURIFICATION.