GENERATION OF ANTIGEN-SPECIFIC MABS USING SCID-HU MICE

THE OVERALL GOAL OF THIS WORK IS TO DEVELOP AN ADOPTIVE IMMUNIZATION SCHEME USING SCID MICE RECONSTITUTED WITH HUMAN PERIPHERAL BLOOD LYMPHOCYTES TO GENERATE ANTIGEN-SPECIFIC HUMAN MABS, SCID MICE WILL BE PRE-TREATED WITH CYTOTOXIC AGENTS TO GIVE IMPROVED LEVELS OF EARLY HUMAN CELL RECONSTITUTION. THESE SCID-HU MICE WILL THEN BE IMMUNIZED WITH EITHER SONICATED OR LIVE BORRELIA BURGDORFERI AND HUMAN ANTIBODY RESPONSES MONITORED. SCID-HU MICE PRODUCING ANTIGEN SPECIFIC ANTIBODIES WILL BE SACRIFICED AND THE CHIMERIC TISSUES TRANSFORMED WITH EBV. THIS STEP WILL SELECT OUT HUMAN B LYMPHOCYTES FROM THE MIXTURE, ACTIVATE AND TRANSFORM THEM. LYMPHOBLASTOID LINES PRODUCING ANTIGEN-SPECIFIC HUMAN ANTIBODIES WILL BE STABILIZED AND IMMORTALIZED BY HYBRIDIZATION WITH A MOUSE- HUMAN HETEROMYELOMA LINE. IT IS HOPED THAT THIS WILL LEAD TO A GENERAL METHOD OF HUMAN CELL IMMUNIZATION AND HUMAN MAB FORMATION. THESE HMABS CAN BE UTILIZED AS INTERNATIONAL STANDARDS FOR CLINICAL DIAGNOSTIC ASSAYS.