Gad₆₅ is recognized by t-cells, but not by antibodies from nod-mice

Abstract

Since the 64kDa-protein glutamic acid decarboxylase (GAD) is one of the major autoantigens in T-cell mediated Type 1 diabetes, its relevance as a T-cell antigen needs to be clarified. After isolation of splenic T-cells from non-obese diabetic (NOD) mice, a useful model for human Type 1 diabetes, we found that these T-cells proliferate spontaneously when incubated with human GAD₆₅, but only marginally after incubation with GAD₆₇, both recombinated in the baculovirus system. No effect was observed with non-diabetic NOD mice or with T-cells from H-2 identical NON-NOD-H-2^g7 control mice. It has been published previously that NOD mice develop autoantibodies against a 64kDa protein detected with mouse beta cells. In immunoprecipitation experiments with sera from the same NOD mice and ³³S-methionine-labelled GAD, no autoantibody binding could be detected. We conclude firstly that GAD₆₅ is an important T-cell antigen which is relevant early in the development of Type 1 diabetes and secondly that there is an antigenic epitope in the human GAD₆₅ molecule recognized by NOD T-cells, but not by NOD autoantibodies precipitating conformational epitopes. Our results therefore provide further evidence that GAD₆₅ is a T-cell antigen in NOD mice, being possibly also involved in very early processes leading to the development of human Type 1 diabetes.

Key Words:

• Type 1 (insulin-dependent) diabetes mellitus
• non obese diabetic (NOD) mouse
• autoimmunity
• T-cells
• glutamic acid decarboxylase (GAD)