中文摘要：

摘要反复冻融B16F10肿瘤细胞制备裂解物，以白喉毒素（Diphtheriatoxin，DT）为载体，OK432和来源于结核分枝杆菌（Mycobacteriumtuberculosis）热休克蛋白70（HSP70）第407-426（mHSP70_407~426，M）的两段串联重复序列M2为佐剂，制备了肿瘤细胞疫苗B16F10-DT-M2-0K432（BDTMOK），探讨其能否抑制小鼠B16黑色素瘤，并且对其抗肿瘤的作用机理进行部分探讨。以制备的BDTMOK免疫C57BL／6小鼠，分别检测体液免疫应答和细胞免疫应答。通过ELISA法，从血清中检测到高滴度的抗B16肿瘤细胞裂解物（B16tumorcelllysate，B16TCL）类抗体。淋巴细胞增殖实验的结果显示，BDTMOK的免疫能够有效的刺激脾淋巴细胞的增殖。预防结合治疗性实验的结果显示，BDTMOK激发的免疫应答对于B16肿瘤攻击起到有效的保护作用，与PBS阴性对照组比较，皮下注射BDTMOK可以延长皮下移植瘤发生的潜伏期（P〈0．05），并且平均瘤重显著降低（P〈0．05）；抑制了小鼠皮内肿瘤模型中的血管新生（P〈0．01）。疫苗BDTMOK能有效的抑制小鼠B16黑色素瘤的生长。

英文摘要：

To make tumor vaccine B16F10-DT-M2-OK432,the freeze-thaw method was adopted to obtain B16FI0 tumor cell lysate. Diphtheria toxin（DT） ,OK432 or M2 which was two tandem repeats of sequence 407-426 of microbial HSP70 were selected as carrier and adjuvants respectively. The C57BL/6 mice were immunized with B16F10-DT-M2-OK432 in order to explore whether the tumor cell vaccine can effectively inhibit B16F10 melanoma in tumor-beating mice and to study the mechanism of B16F10-DT-M2-OK432. After the last immunization, humoral immune and cellular immune response were detected. High titer of anti-B16F10 tumor cell lysate antibody was detected in immunized mice sera by ELISA. Splenic lymophocyte proliferation assay results showed that the proliferation activity of splenocytes from mice immunized with B16F10-DT-M2-OK432 vaccine was significantly higher. The results of prophylactic/ therapeutic experiment showed that B16F10-DT-M2-OK432 immunization significantly inhibited the growth of B16F10 tumor in mice. The tumor latency was prolonged and the average tumor weight in the mice immunized with B16F10-DT-M2-OK432 was signifi- cantly lower than that of mice injected with PBS （ P 〈 0.05 ）. Meanwhile, it also attenuated tumor-induced angiogenesis in intradermal tumor model mice（P 〈 0.01 ）. B16F10-DT-M2-OK432 vaccine can effectively suppress the growth of B16F10 melanoma in mice.