目的：研究IL-2与IL-12协同抗肿瘤作用及其基因治疗的可行性及疗效。方法：以逆转录病毒载体介导hIL-2基因转染人胃腺癌细胞，利用MTT绘制IL-2基因转导细胞和亲本胃腺癌细胞的生长曲线，应用转基因细胞MKN45/IL-2与IL-12对荷瘤小鼠进行治疗，以其对小鼠移植瘤的生长速度及对生存期的影响，作为抗肿瘤效果的评价；应用逆转录病毒构建IL-12表达载体，以IL-12基因修饰的肿瘤细胞EL-4/IL-12和IL-2治疗荷瘤小鼠，观察对小鼠移植瘤的抑制作用及对生存期的影响。结果：转染后的MKN45肿瘤细胞与亲本细胞在体外生长能力上无显著性差异（p>0.05）；体内试验表明MKN45/ IL-2和IL-12治疗组小鼠移植瘤生长缓慢，体积明显小于对照组（p<0.05），生存期延长（p<0.05），二者联合应用可提高治疗效果；EL-4/IL-12细胞接种荷瘤小鼠后可明显抑制皮下移植瘤的生长，延长小鼠生存期（p<0.05），并且与IL-2有协同抗肿瘤作用（p<0.05）；而单独应用重组IL-2进行瘤周注射的抗肿瘤效果不显著（p>0.05）。结论：转基因细胞及其表达的细胞因子可以有效的激发机体的抗肿瘤免疫应答。IL-2与IL-12具有协同抗肿瘤作用。

目的：探讨用放射的IL-12和B7-1转基因细胞进行癌疫苗疗法的效果及协同作用。方法：将逆转录病毒载体重组的小鼠IL-12，B7-1基因表达载体分别导入小鼠EL-4胸腺瘤细胞利用转基因细胞治疗观察其抗肿瘤效果。结果：转基因细胞的肿瘤原性较EL-4/Wt，EL-4/Neo组明显降低 (P<0.001). 同时免疫原性明显增强，以60Co照射的EL-4/IL-12,EL-4/B7-1细胞免疫后，小鼠体内诱发了抗EL-4/Wt的系统性、保护性免疫。用60Co灭活的转基因细胞作为瘤苗进行实验性治疗，能够延长小鼠的生存时间，减慢肿瘤的生长速度，EL-4/IL-12和EL-4/B7-1联合应用较单一的转基因细胞提高了治疗效果 (P<0.01)。结论：放射的转基因细胞及其表达的IL-12细胞因子可以有效地激发机体的抗肿瘤免疫应答，IL-12与B7-1联合应用可以成为一种很有前景的癌疫苗。

Objective: To compare the difference in proliferation and cytotoxicity of A-NK cells cultured with serum-free medium AIMV and standard serum-containing medium in vitro, and also observe the assist effect of IL-12 on A-NK/IL-2 treatment and the morphology character of NK cells. Methods: Cellular proliferation was compared by MTT method in vitro. The morphology of the target cells killed by adherent natural killer cells (A-NK) was observed through electroscope. Result: All of the A-NK cells cultured in different conditions could rapidly proliferate (p>0.05) and kill target cells (p>0.05), death patterns of tumor target cells were necrosis and apoptosis. Conclusion: These data indicate that serum-free medium AIMV could replace standard serum-containing medium for culturing A-NK cells in vitro, A-NK cells activated by both IL-2 and IL-12 was superiority to the IL-2 activated cells, and could reduce side effects by high dose IL-2. The studies provided a new experimental basis for experimental and clinical treatment of A-NK cell in future.

Objective: We have studied the vaccine and synergic effects of IL-12 and B7-1 transfectant on antitumor immunity in vivo. Methods: The retrovirus vector encoding mIL-12 and mB7-1 gene was tranfected into EL-4 thymic lymphoma cells respectively. The cells were used to treated as the tumor vaccine and the therapeutic effect was observed. Results: In contrast to the mice immunized with EL-4/Wt or EL-4/Neo groups, the tumorigenicity of EL-4/IL-12 transfectant was decreased (p<0.001). The EL-4/IL-12 and EL-4/B7-1 cells irradiated with 60Co showed significant systematic protective effects against the sequence rechallenge of EL-4/Wt. EL-4/IL-12 cells with 60Co irradiated delayed the occurrence of tumor and prolonged the survival period of tumor bearing mice. Combination of the vaccines of EL-4/IL-12 and EL-4/B7-1 result in the enhanced the therapeutic effect compaired with each single transfectant group (p<0.001). Conclusion: The results showed that IL-12 transduced cells could enhance the antitumor immunity of host as cancer vaccine. Combination of the EL-4/IL-12 and EL-4/B7-1 transfectant could improve immunity of host and is a prospect Cancer vaccine.

Objective: We have studied the vaccine and synergic effects of IL-12 and B7-1 transfectant on antitumor immunity in vivo. Methods: The retrovirus vector encoding mIL-12 and mB7-1 gene was tranfected into EL-4 thymic lymphoma cells respectively. The cells were used to treated as the tumor vaccine and the therapeutic effect was observed. Results: In contrast to the mice immunized with EL-4/Wt or EL-4/Neo groups, the tumorigenicity of EL-4/IL-12 transfectant was decreased (p<0. 001). The EL-4/IL-12 and EL-4/B7-1 cells irradiated with 60Co showed significant systematic protective effects against the sequence rechallenge of EL-4/Wt. EL-4/IL-12 cells with 60Co irradiated delayed the occurrence of tumor and prolonged the survival period of tumor bearing mice. Combination of the vaccines of EL-4/IL-12 and EL-4/B7-1 result in the enhanced the therapeutic effect compaired with each single transfectant group (p<0. 001). Conclusion: The results showed that IL-12 transduced cells could enhance the antitumor immunity of host as cancer vaccine. Combination of the EL-4/IL-12 and EL-4/B7-1 transfectant could improve immunity of host and is a prospect Cancer vaccine.