| Abstract
| - The non-obese diabetic (NOD) mouse spontaneously develops T-cell-mediated autolmmune Insulitis.We analyzed the clonotypes of Tcell inflltrates of the NOD mouse Islets using a new method we have developed recently, which consists of RT-PCR amplification of the CDR3 region of the TCR β chain mRNA and subsequent single-stand conformation polymorphism(SSCP) analysis.NOD mic of 10-32 weeks of age were shown to accumulate ollgocional T cell in the pancreas. To examine wheather each T cell clone stays in small area of the pancreas or spreads over the whole pancreas, a pancreas was divided into two pieces, which werethem subsequently analyzed in a pair by the above PCR-SSCP method. When a pair produces common bands with the same mobility in SSCP gel, they are likely to represent the presence of the same T cell clones between these two parts of the pancreas. Aged mice (24-32 weeks old) with severe Insulitis obvlousy produced more common bands for most of the Vβ subfamilles than younger mice (10 weeks old) with only perlinslitis sequencing verified that these common bands have the same TCR junction sequences, suggeesting that they were derived from the same T cell clones. These results suggest that clonal revalence of T cells infiltrating into the pancreas occurs in the late satage of insulitis development and that a limited number of T cel clones finally predominate over the whole panceas.
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