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A quantitative immunocytochemical method for the measurement of islet cell cytoplasmic antibodies
1Department of Endocrinology and Immunology, Novo Nordisk A/S, Bagsvaerd, Denmark.
A quantitative immunocytochemical method for the measurement of islet cell cytoplasmic antibodies has been developed. The method employs human or rat pancreas, a protein A-peroxidase/diaminobenzidine secondary antibody system and an independent measurement of islet total and exocrine mean integrated absorbance by scanning microdensitometry. Specific islet cell cytoplasmic antibody binding (islet total-exocrine mean integrated absorbance) was dependent on serum dilution and substrate reaction time. The detection limit was approximately 5 JDF units. Specific islet cell cytoplasmic antibody binding values with human and rat pancreas were similar. Specific islet cell cytoplasmic antibody binding (human pancreas) was greater (p less than 0.001) in sera from patients with newly diagnosed insulin dependent diabetes mellitus (0.119 +/- 0.086, n = 29) compared to normal sera (0.003 +/- 0.008, n = 29). Thus, the method has been validated and may be useful for measuring the blocking effect of potential antigens on specific islet cell cytoplasmic antibody.
A quantitative immunocytochemical method for the measurement of islet cell cytoplasmic antibodies has been developed. The method employs human or rat pancreas, a protein A-peroxidase/diaminobenzidine secondary antibody system and an independent measurement of islet total and exocrine mean integrated absorbance by scanning microdensitometry. Specific islet cell cytoplasmic antibody binding (islet total-exocrine mean integrated absorbance) was dependent on serum dilution and substrate reaction time. The detection limit was approximately 5 JDF units. Specific islet cell cytoplasmic antibody binding values with human and rat pancreas were similar. Specific islet cell cytoplasmic antibody binding (human pancreas) was greater (p less than 0.001) in sera from patients with newly diagnosed insulin dependent diabetes mellitus (0.119 +/- 0.086, n = 29) compared to normal sera (0.003 +/- 0.008, n = 29). Thus, the method has been validated and may be useful for measuring the blocking effect of potential antigens on specific islet cell cytoplasmic antibody.