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...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 vs77.68±12.80,43.10 ±10.05 vs 80.02±13.74,42.14±9.72vs 79.10±2.65,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.展开更多
过继免疫治疗（AIT）是肿瘤生物治疗中的主要疗法。20世纪80年代起，LAK、TIL、CIL、CD3AK等效应细胞陆续被研究，国内外基础研究和临床应用报道层出不穷。1988年Vujanovic首次从LAK细胞中分离纯化了一种具有高效杀伤活性的LAK亚群，因...过继免疫治疗（AIT）是肿瘤生物治疗中的主要疗法。20世纪80年代起，LAK、TIL、CIL、CD3AK等效应细胞陆续被研究，国内外基础研究和临床应用报道层出不穷。1988年Vujanovic首次从LAK细胞中分离纯化了一种具有高效杀伤活性的LAK亚群，因其具有黏附在塑料表面的特征，故命名为A-LAK（adherent LAK）。20世纪90年代后以A—NK（IL-2 activated NK cell）代替了A-LAK。本实验参照文献简便快速地制备A-NK细胞，并在动物肝癌模型研究其抗肿瘤作用。展开更多
Objective： To observe the proliferation of A-NK in vitro and antitumor activity in vivo. Methods： The growth curve of A-NK and NA-NK cells was drawn in vitro. In the rat model, we compared the regional administratio...Objective： To observe the proliferation of A-NK in vitro and antitumor activity in vivo. Methods： The growth curve of A-NK and NA-NK cells was drawn in vitro. In the rat model, we compared the regional administration of A-NK-/IL-2 with the systemic administration. Results： The expansion of A-NK cells reached to climax on day 10 in the culture, increased 16.08 folds compared with the only 3.36 folds for NA-NK cells. In the rat model, we found that the regional administration of A-NK/IL-2 was better than systemic administration or administration of NA-NK/IL-2 not only in tumor infiltration and antitumor response, but also in the survival rate of rats （P〈0.05）. Conclusion： A-NK cells is a new immune effecter cells with high expansibility and high antitumor activity in vivo and in vitro.展开更多
To compare the differences between proliferation and cytotoxicity of adherent natural killer （A-NK） cells cultured with serum-free medium AIMV and standard serum-containing medium in vitro, and also observe the assi...To compare the differences between proliferation and cytotoxicity of adherent natural killer （A-NK） cells cultured with serum-free medium AIMV and standard serum-containing medium in vitro, and also observe the assisting effect of IL-12 on the activation and the morphology character of IL-2-treated A-NK cells, cellular proliferation was evaluated by MTT method in vitro. The morphology of the target cells killed by A-NK cells was observed through electroscope. All of the A-NK cells cultured in serum-free medium AIMV could rapidly proliferate and keep high cytotoxicity compared with that in standard serum-containing medium. A-NK cells activated by both moderate-dose IL-2 and IL-12 were superior to the high-dose IL-2-treated A-NK cells. These data indicated that serum-free medium AIMV could replace standard serum-containing medium for culturing A-NK cells, and moderate-dose IL-2 and IL-12 could reduce side effects caused by high-dose IL-2. The study provided a new experimental basis for experimental and clinical preparation of A-NK cells. Cellular ＆ Molecular Immunology.展开更多
文摘目的: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 vs77.68±12.80,43.10 ±10.05 vs 80.02±13.74,42.14±9.72vs 79.10±2.65,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.
文摘过继免疫治疗（AIT）是肿瘤生物治疗中的主要疗法。20世纪80年代起，LAK、TIL、CIL、CD3AK等效应细胞陆续被研究，国内外基础研究和临床应用报道层出不穷。1988年Vujanovic首次从LAK细胞中分离纯化了一种具有高效杀伤活性的LAK亚群，因其具有黏附在塑料表面的特征，故命名为A-LAK（adherent LAK）。20世纪90年代后以A—NK（IL-2 activated NK cell）代替了A-LAK。本实验参照文献简便快速地制备A-NK细胞，并在动物肝癌模型研究其抗肿瘤作用。
基金This work was supported by the National Natural Science Foundation of China(No. 39270765).
文摘Objective： To observe the proliferation of A-NK in vitro and antitumor activity in vivo. Methods： The growth curve of A-NK and NA-NK cells was drawn in vitro. In the rat model, we compared the regional administration of A-NK-/IL-2 with the systemic administration. Results： The expansion of A-NK cells reached to climax on day 10 in the culture, increased 16.08 folds compared with the only 3.36 folds for NA-NK cells. In the rat model, we found that the regional administration of A-NK/IL-2 was better than systemic administration or administration of NA-NK/IL-2 not only in tumor infiltration and antitumor response, but also in the survival rate of rats （P〈0.05）. Conclusion： A-NK cells is a new immune effecter cells with high expansibility and high antitumor activity in vivo and in vitro.
文摘To compare the differences between proliferation and cytotoxicity of adherent natural killer （A-NK） cells cultured with serum-free medium AIMV and standard serum-containing medium in vitro, and also observe the assisting effect of IL-12 on the activation and the morphology character of IL-2-treated A-NK cells, cellular proliferation was evaluated by MTT method in vitro. The morphology of the target cells killed by A-NK cells was observed through electroscope. All of the A-NK cells cultured in serum-free medium AIMV could rapidly proliferate and keep high cytotoxicity compared with that in standard serum-containing medium. A-NK cells activated by both moderate-dose IL-2 and IL-12 were superior to the high-dose IL-2-treated A-NK cells. These data indicated that serum-free medium AIMV could replace standard serum-containing medium for culturing A-NK cells, and moderate-dose IL-2 and IL-12 could reduce side effects caused by high-dose IL-2. The study provided a new experimental basis for experimental and clinical preparation of A-NK cells. Cellular ＆ Molecular Immunology.
