肿瘤细胞裂解物抗小鼠B16F10黑色素瘤的作用研究

反复冻融B16F10肿瘤细胞制备裂解物,以白喉毒素(Diphtheria toxin,DT)为载体,OK432和来源于结核分枝杆菌(Mycobacterium tuberculosis)热休克蛋白70(HSP70)第407-426(mHSP70407～426,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-OK432(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(B16 tumor cell lysate,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。

Anti-Tumor Effects of B16F10 Tumor Cell Lysate Vaccine in Mouse Melanoma

To make tumor vaccine B16F10-DT-M2-OK432,the freeze-thaw method was adopted to obtain B16F10 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-bearing 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 significantly 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.