CDNA OF JANUSINE, A NOVEL IMMUNOMODULATORY MONOKINE

HIV INFECTED MACROPHAGES RELEASE ELEVATED QUANTITIES OF IL1 AND A NOVEL 7KD MONOKINE WITH PLEIOTROPIC ACTIVITIES. ON THE ONE HAND, IT INHIBITS IL1 AND IL6 INDUCED T LYMPHOCYTE COMITOGENESIS, BUT ON THE OTHER HAND, IT STIMULATES B CELL MITOGENESIS. THIS IMMUNOMODULATORY MONOKINE IS CALLED JANUSINE BECAUSE OF ITS SEEMINGLY OPPOSITE BIOACTIVITIES. IT IS HYPOTHESIZED THAT JANUSINE PLAYS AN IMPORTANT ROLE IN THE PATHOGENESIS OF HIV INFECTION AND OTHER CONDITIONS OF DYSFUNCTIONAL IMMUNOREGULATION. THE OVERALL GOAL OF THE RESEARCH IS TO CLONE, EXPRESS, AND CHARACTERIZE THE ACTIVITIES OF JANUSINE. IN PHASE I, THE FULL LENGTH CDNA CLONES OF JANUSINE WILL BE OBTAINED. CDNA LIBRARIES WILL BE CONSTRUCTED IN LAMBDA GT 11 USING MRNA FROM LPS STIMULATED HUMAN MACROPHAGES OR MACROPHAGE CELL LINES AND SCREENED USING A NEUTRALIZING IMMUNOBLOT POSITIVE ANTI-JANUSINE MONOCLONAL ANTIBODY. BIOACTIVE JANUSINE CLONES WILL BE IDENTIFIED USING EXPRESSION IN THE OKAYAMA-BERG VECTOR TRANSFECTED INTO COS 7 CELLS. THE JANUSINE CDNA WILL BE EXPRESSED AND THE PROTEIN PURIFIED FOR BIOLOGICAL STUDIES. IN PHASE II, THE THERAPEUTIC POTENTIAL OF JANUSINE WILL BE EVALUATED. THE CLONING OF JANUSINE MAY PROVIDE A NOVEL THERAPEUTIC FOR DISEASES CHARACTERIZED BY UNCONTROLLED INFLAMMATION SUCH AS RHEUMATOID ARTHRITIS. INHIBITORS OF THIS MONOKINE MAY BENEFIT AIDS OR SEPSIS PATIENTS.