Generation and maintenance of autoantigen-specific CD8(+) T cell clones isolated from NOD mice

The non-obese diabetic (NOD) mouse develops insulin dependent diabetes mell itus (IDDM) spontaneously with a higher incidence in females than in males, There are many similarities to the human disease, making it an ideal model . Our group is examining the role that CD4(+) and CD8(+) T cells play in ID DM in the NOD mouse, as it is known that both T cell subsets are required f or onset of disease. Although IDDM has an autoimmune etiology, the initial triggering event is unknown and the autoantigen involved has not been ident ified. This investigation focussed on one of the potential autoantigens inv olved, the enzyme glutamic acid decarboxylase (GAD). We raised GAD peptide- specific CD8(+) T cells by immunising NOD mice with the GAD peptide alongsi de an irrelevant peptide that induced a CD4+ T cell response. In order to m aintain these peptide specific T cells in vitro and generate clones, it was found that antibodies specific to CD4(+) and MHC class II molecules needed to be included in the culture medium. This paper outlines the methods we e mployed to generate and maintain these CD8(+) T cells in vitro. (C) 1999 El sevier Science B.V. All rights reserved.