IL-2重组毒素的克隆、表达及对IL-2R^+淋巴细胞的特异性细胞毒效应

The cloning,expressing of IL-2-PE40’ chimeric gene and cytotoxic effect on the activated IL-2R^+ lymphocytes

目的 :为了探讨肿瘤细胞的靶向性杀伤途径。方法 :采用PCR Cloning技术和DNA重组技术 ,构建了 2类IL 2 PE40’嵌合基因的表达载体 ,分别由IPTG和温度 (42℃ )诱导表达 ,并对表达产物进行了酶切片段大小、电泳迁移率、分子量、IL 2和PE40’组分抗原性及细胞学效应的检测。结果 :发现重组毒素对表面表达IL 2受体的活化T淋巴细胞具有明显杀伤效应 ,并有抑制混合淋巴细胞反应的作用。结论 :该IL 2 PE40’融合蛋白确实具有明显的特异性免疫抑制效应。

Objective:To find a therapeutic pathway special to kill tumour cells.Methods:The PCR cloning technique was used to construct three prokaryotic expressing vectors into which chimeric IL 2 PE40’ gene fragment was inserted and induced to express by IPTG or 42℃ respectively.The SDS PAGE、ELISA、cytological and enzymological assay were also used to identify the different characteristic of the expressed product.Results:It was found that IL 2 PE40’ fusion protein was very toxic to the activated IL 2R + T lymphocytes and able to inhibit mixed lymphocytes reaction in vitro.Conclusion:The IL 2 PE40’ fusion constructed had obvious inhibition to body immune function.