树突状细胞疫苗诱导免疫效应细胞对肝癌细胞BEL-7402的生长抑制

Immunoeffector cells induced by dendritic cells vaccine for 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(Dendriticcells,DC)诱导免疫效应细胞对BEL-7402的生长抑制。方法:体外分别用人GM-CSF、IL-4、TNF-a、BEL-7402肿瘤相关抗原(TAA)和人IL-2刺激正常人DC和去DC的单核细胞(免疫效应细胞),6d后混合培养DC和免疫效应细胞1d。体外实验时效应细胞分无DC刺激组(A0组)、细胞因子培养的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-7402前2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激活的免疫效应细胞后再间断给予DC培养上清液6次。对照组(Ⅰ)给等量的1640培养液。结果: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12天时12例全部发生移植瘤;组Ⅱ,观察30d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相差有非常显著性意义(P=0.00466);于给予DC激活的免疫效应细胞后的第45天处死所有裸鼠并称瘤体的重量,组Ⅰ、Ⅱ、Ⅲ两两比较相差有非常显著性意义(P<0.01)。结论:DC在抗恶性肿瘤中有重要作用。

Objective： To observe in vitro and vivo multi-cytokines or tumor allogenic antigen （TAA） stimulated normal dendritic cells （DC） to induce immunoeffect cells on inhibiting the growth of human hepatocellcular cancer cell line BEL-7402. Methods： The normal DC and mononuclear cells without DC were stimulated with combination of granulocyte/macrophage colony stimulating factor, interleukin-4 and tumor necrosis factor-a and TAA In vitro DC were cultured with mononuclear cells without DC （immunoeffector cells） with interleukin-2 （100 U/mL） for 6 days, then mix-cultured DC and immunoeffector cells for one day. The immunoeffector cells were divided into A0 group （DC stimulated without culture）, A1 group （DC stimulated, cultured with cytokines）, A2 group （DC stimulated, culturedwith cytokines and TAA） in vitro study. In vivo study, the nude mice were allocated into 3 groups： preventive group （Ⅱ）, 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 6 times; control group （Ⅰ） gave equivalents amount of 1640 culture liquid. Results： The maximal killing rates were 3.5%, 68.1% and 81% in A0, A1 and A2 groups respectively in vitro. In vivo study, all 12 nude mice developed implanted tumor on the 12th day in group Ⅰ and group Ⅲ; in group Ⅱ, on the 60th and 45th day, one out of the six nude mice developed implanted tumor. Comparing group Ⅰ and Ⅱ, the statistical difference was very significant （P=0.004）; on given DC vaccine stimulated immunoeffector cells at the 45 th day, safrificed all the nude mice and weighed the tumor body, the difference in group Ⅰ, Ⅱ and Ⅲ was significant （P 〈 0.01）. Conclusion： DC vaccine may play an important role in the prevention and treatment of malignant tumor.