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目的探讨人血管内皮细胞生长因子受体-2(KDR)胞外段(Ig1~3)mRNA修饰的树突状细胞(DC)诱发的体外特异性细胞免疫效应。方法构建pmRNA IRES-hKDR)(Ig1~3),体外转录出相应的mRNA,Western blot鉴定其蛋白表达;以hKDR(Ig1~3)mRNA致敏来源于小鼠骨髓的DC,并免疫小鼠,同时设空白DC对照组,7d后取鼠脾细胞为效应细胞,表达mKDR的肿瘤细胞为靶细胞按100∶1、50∶1、25∶1的比例混合,用乳酸脱氢酶(LDH)释放法检测细胞毒作用。结果未经修饰DC激活的细胞毒性T淋巴细胞(CTL)对靶细胞的杀伤率为(19.6±3.7)%、(10.2±2.6)%、(6.2±2.2)%,经mRNA修饰的DC激活的CTL对靶细胞杀伤率为(75.2±2.3)%、(54.3±3.4)%、(20.9±3.1)%,同一效靶比情况下显著强于其他3组。结论hKDR(Ig1~3)mRNA致敏的DC能诱导针对Hepa1-6(mKDR)的特异性细胞免疫反应。
Objective To investigate the in vitro specific cellular immune response induced by dendritic cells (DCs) modified by extracellular domain (Ig1 ~ 3) of human vascular endothelial cell growth factor receptor-2 (KDR). Methods The mRNA of IRES-hKDR (Ig1-3) was constructed and the corresponding mRNA was transcribed in vitro. The protein expression was detected by Western blot. The DCs derived from bone marrow of mice were sensitized with hKDR (Ig1 ~ 3) mRNA and the mice were immunized. At the same time, a blank DC control group was established. After 7 days, spleen cells were taken as effector cells. The target cells expressing mKDR were mixed in the ratio of 100: 1, 50: 1 and 25: 1 and released with lactate dehydrogenase Method to detect cytotoxicity. Results The cytotoxicity of untreated DCs on CTLs was (19.6 ± 3.7)%, (10.2 ± 2.6)% and (6.2 ± 2.2)%, respectively. The number of cytotoxic T lymphocytes activated by mRNA was The cytotoxicity of CTL to target cells was (75.2 ± 2.3)%, (54.3 ± 3.4)% and (20.9 ± 3.1)%, respectively. Conclusions DC sensitized with hKDR (Ig1 ~ 3) mRNA can induce specific cellular immune responses against Hepa1-6 (mKDR).