目的：多方面比较无血清培养基AIMV与完全培养基对体外细胞因子激活的NK细胞的支持作用。 方法：分别用AIMV及完全培养基培养粘附NK细胞（A-NK）和非粘附NK细胞（NA-NK)，比较细胞的增殖能力、杀伤肿瘤细胞的能力、表达粘附分子CD11C的能力，并通过电镜对A-NK细胞所杀伤的肿瘤细胞进行形态学观察。结果：与完全培养基培养细胞相比，AIMV培养细胞增殖能力、杀伤肿瘤细胞的能力、表达CD11C的能力与之相当；在同一培养条件下A-NK细胞的增殖能力、杀伤活性及表达CD11C的能力明显强于NA-NK细胞；被A-NK细胞杀伤的肿瘤细胞死亡形式是溶解坏死和凋亡。结论：无血清培养基可替代完全培养基用于A-NK细胞的培养；A-NK细胞用于治疗肿瘤的过继免疫疗法要优于NA-NK细胞。

目的：探讨和总结对肿瘤患者进行A-NK细胞/IL-2临床静脉输注的护理经验。方法：观察接受治疗的患者体温、呼吸、血压等生命体征的变化情况。结果：收治患者中除个别人有一过性发热、头痛、呼吸加快的症状外，绝大多数患者无特殊反应，经处理后，症状在短期内消失。结论：应用A-NK细胞/ IL-2静脉输注治疗癌症患者是比较安全的，所采取的护理方法具有简单和实用性。

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.

目的：观察A-NK细胞在体外的增殖和体内抗肿瘤活性。方法：绘制A-NK细胞体外生长曲线，在动物实验中比较了局部和全身投给A-NK细胞的抗肿瘤活性。结果：在培养中，A-NK细胞在第10天获得最大增殖，其扩增数目为16.08倍，而NA-NK细胞仅为3.06倍，在动物实验中，局部投给A-NK细胞在肿瘤侵润和抗肿瘤效果以及生存期方面均好于NA-NK细胞(P<0.05)。结论：A-NK细胞是一种新型的免疫效应细胞，在体内外具有较高的增殖和抗肿瘤活性，此研究为A-NK细胞进行肿瘤临床的过继免疫治疗提供了新的方法学。

目的：比较A-NK细胞在完全培养基（CM）和无血清培养基 (AIMV) 中体外扩增及杀伤肿瘤细胞作用，同时探讨IL-12对于A-NK/IL-2治疗的辅助作用及其杀伤的形态学观察。 方法：用MTT法比较不同培养条件下的A-NK细胞体外增殖能力，并测定细胞体外杀伤肿瘤细胞的活性。通过做扫描和透射电镜，对A-NK细胞杀伤的肿瘤细胞进行形态学观察。结果：不同培养条件下的A-NK细胞均可在短期内大量扩增（p<0.05）,且均有很强的杀伤肿瘤的活性（p<0.05）。被A-NK细胞杀伤的肿瘤细胞的死亡形式是溶解坏死（necrosis）和凋零坏死(apoptosis)。结论：AIMV可替代CM用于A-NK细胞的培养。联合应用IL-12和IL-2激活A-NK细胞优于单独使用IL-2，可减轻高剂量IL-2带来的副作用。本研究为A-NK细胞的实验室研究和将来临床应用推广奠定了新的实验基础。

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.

目的：用IL-4联合IL-2共同诱导A-NK细胞，研究A-NK细胞体外细胞毒性及对直肠肿瘤生长的抑制作用。方法：用基因重组的IL-2和IL-4联合活化A-NK细胞，用LDH-L释放法测定效应细胞的细胞活性，同时观察A-NK细胞对人直肠癌细胞在裸鼠体内的生长抑制作用。结果：IL-2单独或IL-2联合IL-4均可以诱导A-NK细胞，但二者联合效果更好，通过LDH-L测定活化的A-NK细胞可在体外杀伤K562，Anip973和CC95细胞（39.00 ± 9.16 vs 77.68 ±12.80，43.10 ± 10.05 vs 80.02 ± 13.74，42.14 ± 9.72 vs79.10 ±12.65，P<0.01)，抑制人直肠癌肿瘤在裸鼠体内的生长（1.04 ± 0.15 vs 0.62 ± 0.16，P<0.01)，提示A-NK细胞膜表面存在IL-4受体的表达。结论：IL-4能够增强IL-2诱发的A-NK细胞抗肿瘤活性，此方法在将来肿瘤临床治疗中具有潜在的应用价值。

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.

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。

白细胞介素12是近年来研究最热门的一种细胞因子，有着多彩的生物学活性，在T细胞和NK等细胞的诱导、活化，免疫功能的调节，抗细菌、抗病毒、抗肿瘤和自身免疫病等方面发挥着非常重要的作用。本文仅围绕着它的结构，生物学活性和抗肿瘤方面的最新进展作以简要介绍。