不同刺激诱导肺癌患者引流淋巴结细胞抗自体肿瘤的体外研究

The study of antitumor effect of tumor-draining lymph node cells from lung cancer patients induced by different stimulus in vitro

目的:探索适合于临床大规模应用的有效刺激、活化肿瘤引流淋巴结(Tumor-draininglymphnode,TDLN)细胞的方法。方法:对3种刺激剂(IL-2、IL-2+自身肺癌细胞抗原和IL-2+GM-CSF+IL-4+自身肺癌细胞抗原)诱导TDLN细胞体外抗自体肿瘤作用,通过乳酸脱氢酶法测定了CTL杀伤活性,观察了细胞形态学变化,通过流式细胞技术测定了细胞CD83的阳性表达率。结果:经MTT法检测,IL-2+GM-CSF+IL-4+自身肺癌细胞抗原诱导的TDLN细胞的增殖程度明显高于IL-2+自身肺癌细胞抗原刺激组和IL-2刺激组(P<0·01)。IL-2+GM-CSF+IL-4+自身肺癌细胞抗原诱导的肿瘤特异性CTL活性明显高于IL-2+自身肺癌细胞抗原刺激组和IL-2刺激组,CD83阳性表达细胞率也明显高于IL-2+自身肺癌细胞抗原刺激组和IL-2刺激组。结论:用IL-2+GM-CSF+IL-4+自身肺癌细胞抗原刺激和活化TDLN,优于单独用IL-2刺激或IL-2+自身肺癌细胞抗原刺激,该作用可能与其诱导TDLN细胞产生DC数量的增加有关。

Objective：To find a feasible method to stimulate tumor-draining lymph node （TDLN） ceils in clinic. Methods：CTL activity of TDLN ceils induced by different stimulus （ IL-2 group, IL-2 ＋ autolngous tumor antigen group, IL-2 ＋ GM-CSF ＋ IL-4 ＋ autologous tumor antigen group） was measured by the method of maximal LDH enzyme release. The mechanisms were explored by observation in morphology and detection of the CD83 positive rate of TDLN cells. Results ：The level of growth of TDLN cells induced by （IL- 2 ＋ GM-CSF ＋ IL-4 ＋ autologous tumor antigen） was significantly higher than TDLN ceils induced by IL-2 and （ IL-2 ＋ autolngous tumor antigen） （ P 〈 0. 01 ）. Antitumor CTL activity of TDLN ceils induced by IL-2 ＋ GM-CSF ＋ IL-4 ＋ autologous tumor antigen was significantly higher than the two other groups. The CD83 positive rate of TDLN ceils induced by IL-2 ＋ GM-CSF ＋ IL-4 ＋ autologeus tumor antigen was significantly heightened. Conclusion ：The method of stimulating tumor-draining lymph node （TDLN） ceils by （IL-2 ＋ GM- CSF ＋ IL-4 ＋ autologeus tumor antigen） is better than by IL-2 or （ IL-2 ＋ autologeus tumor antigen）. The results might be assotiate with the facts that TDLN ceils induced by IL-2 ＋ GM-CSF ＋ IL-4 ＋ autologous tumor antigen could produced more dendritic ceils（DC）.