ANTIBODY-MEDIATED DETECTION SYSTEMS FOR ACROLEIN-DNA ADDUCTS

THE LONG-TERM OBJECTIVES OF THIS PROJECT ARE (1) TO ESTABLISH A STABLE SOURCE OF WELL-CHARACTERIZED ANTIBODIES TO DNA ADDUCTS OF CARCINOGEN/MUTAGENS SO THAT THESE IMPORTANT PROBES CAN BE MADE AVAILABLE FOR FUNDAMENTAL RESEARCH INTO THE MECHANISM(S) OF CARCINOGENESIS, AND (2) TOPROVIDE SERVICES/ASSAY KITS FOR MONITORING HUMAN POPULATIONSFOR POSSIBLE EXPOSURE TO SOME ENVIRONMENTAL CARCINOGENS BY SCREENING FOR NEOANTIGENS IN BIOLOGICAL SPECIMENS. SPECIFICAIMS FOR THIS PHASE I EFFORT WILL INCLUDE THE PRODUCTION OF ACROLEIN-DNA ADDUCTS BY TREATING CALF THYMUS DNA IN VITRO WITH VARYING CONCENTRATIONS OF THE CHEMICAL TO OBTAIN DNA SAMPLES CONTAINING DIFFERENT LEVELS OF REACTION. THE PROJECT WILL ALSO INCLUDE UTILIZATION OF A NUMBER OF IMMUNOLOGICAL METHODS THAT WOULD REDUCE THE AMOUNT OF IMMUNOGEN OR INCREASE THE EFFICIENCY OF ANTIBODY PRODUCTION BY THE USE OF RBF/DN MICE WITH A MARKER CHROMOSOME THAT CONTAINS THE GENES FOR BOTH APRT AND IMMUNOGLOBIN HEAVY CHAIN; THE USE OF HL-653 CELLS DEFICIENT IN BOTH HPRT AND APRT AS THE FUSION PARTNER; USE OF IN VITRO IMMUNIZATION WITH OR WITHOUT PROTEIN COUPLING; USE OF INTRASPLENIC IMMUNIZATION, OR USE OF ALTERNATE ADJUVANT SYSTEM.