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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。
The B16F10 tumor cells were lyophilized repeatedly to prepare lysates. Diphtheria toxin (DT) was used as a carrier, and OK432 and HSP70 407-426 (mHSP70407 ~ 426), Mycobacterium tuberculosis M) were adjuvanted with adjuvant to establish a tumor cell vaccine B16F10-DT-M2-OK432 (BDTMOK) to investigate whether it can inhibit mouse B16 melanoma and to investigate its anti-tumor mechanism Discussion. The prepared BDTMOK was used to immunize C57BL / 6 mice to detect the humoral immune response and cellular immune response, respectively. High titer anti-B16 tumor cell lysate (B16TCL) antibody was detected from the serum by ELISA. The results of lymphocyte proliferation experiments show that the immunization of BDTMOK can effectively stimulate the proliferation of spleen lymphocytes. The results of combination therapy and prophylaxis showed that BDTMOK elicited an effective protective effect against B16 tumor challenge. Compared with PBS-negative control group, subcutaneous injection of BDTMOK prolonged the incubation period of subcutaneous xenografts (P <0.05), and Mean tumor weight was significantly lower (P <0.05); angiogenesis was inhibited in the mouse intradermal tumor model (P <0.01). Vaccine BDTMOK can effectively inhibit the growth of mouse B16 melanoma.