Reactivity of islet cell antibodies (ICA) to recombinant glutamic acid decarboxylase (GAD)

To study the heterogeneity of islet cell antibodies (ICA), recombinant rat and human GAD65 expressed as bacterial fusion proteins were used to inhibit ICA reactivity in sera from recent onset type 1 diabetic children and ICA-positive first degree relatives of diabetic patients. Rat GAD65 was expressed as a fusion protein in the expression vector RSET and inhibited ICA (GAD+ICA) in 26% of 23 recent onset patients, 29% of 14 ICA positive first degree relatives (FDR) who progressed to diabetes (prediabetics) and 50% of 20 FDR who did not progress to diabetes 18 months to 5 years (31+/-14 months) after collection of the sample. GAD+ICA were inversely associated with the presence of insulin autoantibodies (IAA) (P=0.006). GAD antibodies (GAD-Ab) were also detected by immunoprecipitation of in vitro transcribed and translated [S-35] methionine-labelled human GAD65. GAD-Ab were present in 83% of recent onset patients, 86% of prediabetics and 95% of the relatives who did not progress to diabetes. The level of GAD-Ab was higher in the presence of GAD+ICA (1.39+/-0.57 vs 0.79+/-0.6 index units; P=0.001). ICA levels were higher in GAD-Ab negative than in GAD-Ab positive sera (377 +/- 256 vs 195 +/- 231 JDFU; P=0.03). Our results confirm that a recombinant GAD65 fusion protein can be used to detect ICA heterogeneity. However, neither inhibition of ICA with recombinant GAD, nor direct detection of GAD-Ab improved the prediction of progression to clinical diabetes in ICA positive FDR. (C) 1995 Academic Press Limited