草鱼、中华鳖脾细胞培养上清液IL-2物质的检测

IDENTIFICATION OF AN INTERLEUKIN-2 SUBSTANCE IN SPLENOCYTE CULTURE SUPERNATANT OF GRASS CARP CTENOPHARYNGODON IDELLUS AND CHINESE SOFT-SHELLED TURTLE TRIONYX SINENSIS

    摘要
  • 摘要: 用刀豆蛋白A(ConA)刺激诱导草鱼和中华鳖脾细胞,收集细胞培养上清液,用小鼠胸腺细胞增殖试验和对小鼠L929细胞系杀伤试验检测上清液中白细胞介素2(简称IL-2)活性.结果表明:草鱼、中华鳖脾细胞培养上清液中有IL-2样活性物质,这种物质使小鼠的胸腺细胞3HTdR掺入量明显增加,在相同效靶比的条件下对小鼠L929细胞系杀伤率也显着增强,这种IL-2活性均能被抗人rIL-2血清所抑制.中华鳖脾细胞培养上清液(含IL-2)对中华鳖胸腺细胞也有较明显的促增殖作用并能消除兔抗中华鳖胸腺细胞血清(RATTS)对中华鳖胸腺细胞增殖的抑制作用.进一步用抗人白细胞分化抗原CD25(IL-2受体,简称IL-2R)单克隆抗体进行免疫组化交叉反应提示草鱼、中华鳖淋巴细胞膜上含有与人类IL-2R类似功能和结构的物质.

     

    Abstract: Interleukin 2(IL-2)activity by thymocyte(C57BL/6-8 mice)proliferation assay and fibroblast(L929fibroblasts)cytotoxicity assay has been found in supernatant of mitogen(ConA)stimulated splenocyte cultures of grass carp Ctenophaxyngodon idellus, Chinese soft shelled turtle Trionyx sinensis.This IL-2 activity was neutralized by polyclonal anti human IL-2 antisera.Besides, the splenocyte culture supernatant(IL-2)of Trionyx sinensis increased obviously thymocyte(Trionyx sinensis)proliferation,which was also neutralized by polyclonal anti human IL-2 antisera,and eliminated the suppressive effect of rabbit anti thymocyte(Trionyx sinensis)antisera(RATTS)on thymocyte(Trionyx sinensis).Further,with indirect immunohistochemistry technique,CD25(IL-2 receptor or IL-2R)cross reaction has been identified on lymphocyte membrane of Ctenophaxyngodon idellus and Trionyx sinensis using mouse monoclonal antibody directed against Human Leucocyte Differentiation Antigen CD25(IL-2R).

     

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