树突状细胞疫苗诱导免疫效应细胞对PC3生长的抑制作用

Inhibitory effect of Immunoeffector cells on growth of PC3 induced by dendritic cell vaccines

目的体内外观察多种细胞因子及肿瘤相关抗原(TAA)体外培养的树突状细胞(DC)诱导免疫效应细胞对PC3的抑制。方法用淋巴细胞分离液分离抗凝新鲜全血以获得单核细胞(PBMC),用贴壁法获取DC和去DC的单核细胞(免疫效应细胞),并分别用粒/巨噬细胞集落刺激因子(GM-CSF)、人白细胞介素(IL)-4、肿瘤坏死因子(TNF)-α、PC3肿瘤相关抗原(PC3TAA)和人白介素2(IL-2)培养正常人DC和免疫效应细胞,5-6d后将DC和免疫效应细胞混合培养1-2d。观察DC,免疫效应细胞生长状况。酶法检测DC诱导免疫效应细胞对PC3的毒性作用。体内观察DC诱导免疫效应细胞和DC培养上清液对裸鼠PC3移植瘤的生长抑制作用。结果体外多种细胞因子和肿瘤相关抗原能有效引起DC增殖。DC疫苗诱导免疫效应细胞对PC3的最大杀伤效率为62.4%。体内实验见:阴性对照组Ⅰ,单纯细胞治疗组Ⅲ及其联合培养上清液治疗组Ⅳ裸鼠均于第12天发生移植瘤;预防治疗组Ⅱ,观察30d时,6例只有2例发生移植瘤;于移植瘤产生后的第45天处死所有裸鼠并称瘤体的重量,各组比较差异有统计学意义(P<0.05),两两比较,组Ⅰ分别与组Ⅱ、Ⅲ比较差异有统计学意义(P<0.001),组Ⅰ与组Ⅳ、组Ⅱ与组Ⅲ比较差异有统计学意义(P<0.05)。结论DC疫苗在抗恶性肿瘤中有重要作用。

Objective To observe the inhibition of immunoeffector cells on the growth PC3 cells induced by dentritic ceils （DC） in vitro cultured with multi-cytokines or tumor allogentic antigen （TAA）. Methods In vitro, human GM-CSF, IL-4, TNF-α, PC3 TAA and human IL-2 were use to stimulate normal DC and monouclear cells （immunoeffecetor cells）. Five-6 days later, the DC and immunoeffector cells were cultured for 1-2 days and supernatant harvested separately. In the in vitro cytotoxin study, the immunoeffector cells were divided into A0 group （without DC stimulated）, A1 group （DC stimulated, cultured with cytokines）, A2 group （DC stimulated, cultured with cyto-kines and TAA）. During the in vivo study, the nude mice were allocated to 4 groups： preventive group （Ⅱ）, 1-2 days before inoculating PC3, giving DCactivated immunoeffector cells;treated group （Ⅲ）, only giving DC activated immunoeffector cells after emergence of implanted tumor in all nude mice; treated group （Ⅳ ）, giving DC activated immunoeffector cells after emergence of implanted tumor in all nude mice with intermittent administration of DC cultured supernatant 3-6 times; control group （Ⅰ）, giving equivalent amount of 1640 cultured liquid. Results In vitro, the maximal killing rate was 62.4%. In the in vivo study, all 18 mice developed implanted tumor on the day 12 in group Ⅰ , Ⅲ and Ⅳ; In group Ⅱ, on the 30th day, 2 of the 6 mice developed implanted tumor. After observation for 45 days, only 2 mice had implanted tumor. On the 45th day after development of implanted tumor, all the nude mice were sacrificed and the tumor body weighed. There was significant difference among group Ⅰ, Ⅱ , Ⅲand Ⅳ（P 〈 0.05）. Conclusion DC vaccines may play an important role in anti-tumor therapy.