酵母表面展示IGRP_(206-214)-H-2K^d单链三聚体及对NOD鼠CD8^+T细胞的活化作用

Displaying of IGRP_(206-214)-H-2K^d SCT on yeast surface and activation of CD8^+T cells of NOD mice

通过酵母展示表达IGRP206-214-H-2Kd单链三聚体,筛选诱导条件获得高表达的IGRP206-214-H-2Kd单链三聚体酵母用于研究该三聚体诱导NOD鼠CD8+T细胞活化的作用。结果表明:转染IGRP206-214-H-2Kd单链三聚体的真核表达载体的酵母,在含2%D-半乳糖的酵母培养基中,20℃诱导18h,有77.1%的酵母能稳定表达IGRP206-214-H-2Kd单链三聚体蛋白。展示酵母与NOD鼠脾脏细胞共培养24hCD69+表达量最高,占CD8+T细胞的1.89%,共培养96h,CD25+表达量最高,占CD8+T细胞的4.25%,说明酵母展示获得的酵母能稳定表达IGRP206-214-H-2Kd单链三聚体蛋白对NOD鼠CD8+T细胞具有活化作用。

In this study, we constructed a yeast expression system for IGRPz06_214-H-2Ka-SCT displayed on the yeast cell surface, and screened the yeast with high expression of IGRP206-214-H-2Kd-SCT, to confirm the effect of IGRP206 214-H-2Kd-SCT on CD8＋ T cells of NOD mice. The results showed that, after screened inducing expression condi- tion, 77.1% of yeast cells expressed 1GRP206_214-H-2Kd protein under condition of 2% of D-galactose and 18 h of induced period at 20℃. Then, we proved that with the NOD mouse spleen cells co cultured for 24 h CD69 expression was the highest, accounting for 1.89 % of CDS＋T cells and co cultured for 96 h, CD25＋ expression was the highest, accounting for 4.25% of CD8＋ T cell. Our results suggest that yeast expressed specific proteins could activate spleen CD8＋ T cells of NOD mouse in vitro.