白细胞介素2-重组毒素对活化T淋巴细胞的靶向杀伤效应

利用ＰＣＲ－Ｃｌｏｎｉｎｇ技术和ＤＮＡ重组技术，构建了两类ＩＬ－２－ＰＥ４０＇嵌合基因的表达载体，分别由ＩＰＴＧ和温度（４２℃）诱导表达。在对表达产物进行了酶切位点和片段大小、电泳迁移率、分子量的初步鉴定后，又对融合蛋白的白细胞介素２（ＩＬ－２）和ＰＥ４０＇组分的抗原性进行了ＥＬＩＳＡ鉴定。从菌体蛋白中初步分离纯化了重组毒素，并测定其对细胞表面表达ＩＬ－２受体的活化Ｔ淋巴细胞的靶向杀伤效应及对混合淋巴细胞反应的抑制作用。结果表明，ＩＬ－２－ＰＥ４０＇融合蛋白确实对ＩＬ－２Ｒ＋细胞具有明显的靶向杀伤效应并呈剂量依赖关系，与对照组相比，随着保温时间的延长（６、２４、４８ｈ），杀伤效应趋于增强（Ｐ＜０．０５）。混合淋巴细胞培养结果显示，该融合蛋白对单相和双相反应均有不同程度的免疫抑制作用（Ｐ＜０．０１）。

The Targeted Killing Effect of Recombination Pseudomonas Exotoxin (IL-2-PE40') on Activated and IL-2R+ T-lymphocytes

Two type of prokaryote express vectors into which chimeric IL2PE40' gene fragment was inserted were constructed by using PCRcloning technique and DNA recombinant technique and expressed after addition of IPTG or change of temperature (up to 42 ). Furthermore the fusion proteins expressed in host cells were also identified by using SDSPAG electrophoresis and immunological ELISA. The experimental results showed that expressing products of chimeric gene containing both IL2 and PE antigen components and the expressed IL2PE40' fusion protein was very toxic to the activated IL2R T lymphocytes (compared with the control group, P<005) and able to inhibite the reaction of mixed lymphocytes with signal or double direction in vitro(P<001)