树突状细胞诱导免疫效应细胞对BEL-7402的生长抑制

Immunoeffector cells induced by dendritic cells inhibiting growth of BEL-7402

目的 体内外观察多种细胞因子和 (或 )肿瘤相关抗原 (tumorallogenicantigen ,TAA)刺激的正常人树突状细胞 (DC)诱导免疫效应细胞对BEL 740 2的生长抑制。方法 体外分别用人GM CSF、IL 4、TNFα、BEL 740 2TAA和人IL 2刺激正常人DC和去DC的单个核细胞 (免疫效应细胞 ) ,5～ 6d后混合培养DC和免疫效应细胞 1～ 2d。体外实验时效应细胞分无DC刺激组 (A0组 )、细胞因子培养和DC刺激组 (A1组 )、细胞因子和TAA培养的DC刺激组 (A2组 )。体内实验时 ,裸鼠分为 3组 :预防组 (Ⅱ ) ,于接种BEL 740 2前 1～ 2d天给DC激活的免疫效应细胞 ;治疗组 (Ⅲ ) ,待全部裸鼠移植瘤长出时给DC激活的免疫效应细胞 ;组Ⅱ、Ⅲ于给予DC激活免疫效应细胞后再间断给予DC培养上清液 3～6d。对照组 (Ⅰ )给等量的 164 0培养液。结果 多细胞因子体外可刺激DC发生扩增并高表达B7分子。体外实验中最大杀伤效率 :A2组为 81% ,A1组为 68.1% ,A0组为 3 .5 %。体内实验中 :组Ⅰ和组Ⅲ第 12d时 12例全部发生移植瘤 ;组Ⅱ观察 3 0d时 ,6例有 1例发生移植瘤 ,观察 45d时 ,仍只有 1例发生移植瘤 ,组Ⅰ、Ⅱ相差有非常显著意义 (P =0 .0 0 466) ;在给予DC激活的免疫效应细胞后的第 45d处死所有裸鼠并称瘤体的重量 ,组Ⅰ、Ⅱ、Ⅲ比较相差有非常?

Objective To observe in vitro and vivo multi cytokines or tumor allogenic antigen (TAA) stimulated normal dendritic cells (DC) to induce immunoeffector cells on inhibiting the growth of human hepatocellular carcinoma cell line BEL 7402. Methods In vitro, human GM CSF, IL 4, TNFα, BEL 7402 TAA and human IL 2 stimulated normal dendritic cells and mononuclear cells (immunoeffector cells) without stimulated dendritic cells. 5～6 days later, mixed the culture of DC and immunoeffector cells for 1～2 days. In vitro study, the immunoeffector cells were divided into A0 group (DC stimulated), Al group (DC stimulated,cultured with cytokines), A2 group (DC stimulated, cultured with cytokines and TAA). During the in vivo study, the nude mice were allocated into 3 groups: preventive group (Ⅱ), 1～2 days before inoculating BEL 7402, gave DC activated immunoeffector cells; treated group (Ⅲ), gave 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) gave equivalentes amout of 1640 cultured liquid. Results The multicytokines could stimulate DC to clone and express high B7 molecules in vitro, the maximal killing rate in A2 group was 81%, Al group 68.1% and A0 group 3.5% . In vivo study, all 12 mice developed implanted tumor on the 12th day in group Ⅰ and group Ⅲ; in group Ⅱ, on the 30th day, one out of the six mice develped implanted tumor, after observation for 45 days, still only one mouse had implanted tumor. Comparing group Ⅰ and Ⅱ, the statistical difference was very significant ( P = 0.00466 ); on given DC stimulated immunoeffector cells at the 45th day, safrificed all the nude mice and weighed the tumor body. The differences in group Ⅰ, Ⅱ and Ⅲ was significant ( P < 0.01 ). Conclusion DC may play an important role in the prevention and treatment of malignant tumor.