STANDARDIZATION OF ASSAY FOR ANTIBODIES TO ISLET CELLS

THE TEST FOR ISLET CELL CYTOPLASMIC ANTIBODIES (ICA) IS A GOOD PREDICTOR OF INDIVIDUALS AT RISK TO DEVELOP TYPE I DIABETES AND THUS HAS POTENTIAL VALUE IN IDENTIFYING CANDI- DATES FOR FUTURE MEASURES AIMED AT PREVENTING THE DISEASE. HOWEVER, PROBLEMS WITH STANDARDIZATION LIMIT THE RELIABILITYOF THE ICA ASSAY. THE GOAL OF THIS PROJECT IS TO MAKE AVAILABLE A STANDARDIZED, AND HENCE MORE RELIABLE, SYSTEM FOR DETECTING ICA. SPECIFIC OBJECTIVES ARE TO: (1) ESTABLISH A PROTOCOL FOR PROCURING DONOR PANCREAS THAT RESULTS IN OPTIMAL REACTIVITY AND RETENTION OF TISSUE MORPHOLOGY; (2) ESTABLISH THE MOST SENSITIVE AND REPRODUC- IBLE FLUORESCENT INDICATOR SYSTEM FOR VISUALIZING THE ICA REACTION; (3) MAXIMIZE THE STABILITY OF ISLET CELL ANTIGENS IN TISSUE SECTIONS; AND (4) DEVELOP A SEPARATE PARALLEL SYSTEM THAT ALLOWS THE USER TO ASSESS HIS SKILLS IN PERFORMING, READING, AND INTERPRETING THE ICA TEST. THESE AIMS WILL BE ADDRESSED BY: (1) DOCUMENTING TIME LIMITS FOR TISSUE PROCESSING BEYOND WHICH REACTIVITY AND HISTOLOGY DETERIORATE AND EXAMINING PANCREAS PERFUSION AS A MEANS OF EXTENDING THESE TIME LIMITS; (2) COMPARING FIVE FLOURESCENT INDICATORS FOR SPECIFICITY, SENSITIVITY, AND REPRODUCIBILITYIN THE ICA ASSAY; (3) COMPARING SHELF-LIVES OF PANCREAS SECTIONS AFTER VARIOUS MODES OF FIXATION; AND (4) DEVELOPINGTHE COMPONENTS OF A USER STANDARDIZATION KIT. THIS RESEARCHWILL LAY THE GROUNDWORK FOR PHASE II DEVELOPMENT OF A COMMERCIALLY MARKETABLE ICA ASSAY PACKAGE.