树突状细胞激活效应细胞的最佳效靶比及其体外杀伤活性的研究

STUDY ON THE OPTIMAL EFFICIENT RATIO ON TARGET CELLS AND THE ANTITUMOR ACTIVITY OF EFFECT CELLS STIMULATED BY DENDRITIC CELLS IN VITRO

目的:探讨树突状细胞(DC)激活肿瘤浸润淋巴细胞(TIL)和小鼠脾淋巴细胞(小鼠脾LC)的最佳效靶比并观察被有效激活的TIL和小鼠脾LC等对小鼠H22肝癌细胞的体外杀伤作用。方法:从荷瘤小鼠四肢长骨提取DC,联合应用GM-CSF、IL-4和小鼠H22肝癌细胞全细胞性抗原致敏DC,然后依据不同的激活效靶比用致敏DC体外激活TIL、小鼠脾LC,分别检测、比较被不同程度激活的TIL、小鼠脾LC以及未激活的小鼠脾LC、未激活的TIL对小鼠H22肝癌细胞的杀伤活性。结果:①当DC∶靶细胞(E/T)为1∶400和1∶200时,所激活的TIL或小鼠脾LC的杀伤活性较弱;当E/T为1∶100时,杀伤活性有较明显的提高,与前两者比较,差异有统计学意义(P<0.05);当E/T=1∶50时,杀伤活性增加,与前三者比较差异均有统计学意义(P<0.01或P<0.05);而当E/T=1∶25、1∶12.5和1∶6.25时,杀伤活性与E/T=1∶50时比较无明显变化(P>0.05);②当E/T=1∶50时,未经DC激活的小鼠脾LC(A组)杀伤率为(9.73±1.40)%,未经DC激活的TIL(B组)和DC激活的小鼠脾LC(C组)杀伤率分别为(50.91±2.36)%和(49.70±2.70)%,DC激活的TIL(D组)的杀伤率为(73.49±2.46)%。A组与其他组分别比较,差异均有统计学意义(P<0.01);B组和C组比较差异无统计学意义(P>0.05);D组与其他各组分别比较,差异均有统计学意义(P<0.01)。结论:①当E/T=1∶50时,DC能很好的发挥抗原提呈作用,使小鼠脾LC和TIL得以充分激活;②充分激活的小鼠脾LC或TIL对小鼠H22肝癌细胞的体外杀伤活性较未激活的小鼠脾LC或未激活的TIL均具有明显的提高,尤以激活的TIL的杀伤活性最强。

Abstract Objective： To investigate the optimal efficient ratio of efficacy to target （E/T） and the antitumor activity of tumor-infiltrating lymphocytes（TILs） and the mouse＇s splenocyte lymphocytes stimulated by dendritic cells（DCs） on H22 cells in vitro. Methods： DCs were isolated from mouse bone marrow and stimulated by granuloeyte/ macrophage colony stimulating factor （GM-CSF）, interleukin-4 （IL-4） and tumor cellular antigen. Then TILs and the mouse＇s splenocyte lymphocytes were stimulated by the activation DCs under different E/T conditions, and their anti-tumor activity on H22 cells in vitro were observed respectively. Results：（1）At E/T of 1 ： 400 and 1 ： 200 respectively, the anti-tumor activity of TILs or the mouse＇s splenocyte lymphocytes stimulated by DCs were low respectively; at E/T of 1 ： 100, the antitumor activity increased, and the difference was significant as compared with those at E/T of 1 ： 400 and 1 ： 200 respectively （ P 〈0.05）; at E/T of 1 ： 50, the anti-tumor activity increased remarkably,and there was significant difference as compared with those at E/T of 1 ： 400 , 1 ： 200 and 1 ： 100 respectively （ P〈 0. 01 or P 〈0.05） ; but at E/T of 1 ： 25, 1 ： 12.5 and 1 ： 6.25 respectively, the anti-tumor activity almost had no change as compared with that at E/T of 1 ： 50（ P 〉0.05）. （2）at E/T of 1 ： 50, the anti-tumor activity of the mouse＇s splenocyte lymphocytes not stimulated by DCs（group A） was（9. 73±1.40）%, while the antitumor activity of the TILs not stimulated by DCs （group B） and the mouse＇s splenocyte lymphocytes stimulated by DCs （group C） were （50.91 ± 2.36） % and （49.70±2.70） % respectively. The antitumor activity of the TILs stimulated by DCs （group D） was（73.49± 2.46）%. the anti-tumor activity o{ group A as compared with those of other three groups respectively had a significant difference （ P 〈0.01）,there wasn＇t a significant difference between the anti-tumor activity of group B and that of group C （ P 0. 05）,the anti-tumor activity of group D as compared with the others respectively had a significant difference （P d0. 01）. Conelusion：（1）At E/T of 1 ： 50,TILs and the mouse＇s splenocyte lymphocytes can be stimulated efficiently by DCs. （2）the anti-tumor activity of TILs and the mouse＇s splenocyte lymphocytes stimulated efficiently by DCs on H22 cells were significantly higher than those of TILs and the mouse＇s splenocyte lymphocytes not stimulated by DCs respectively, and especially for TILs stimulated efficiently by DCs, the anti-tumor activity was highest.