分子佐剂修饰的血管内皮生长因子融合蛋白疫苗抗肿瘤活性研究

发布时间：2018-11-12 07:40

【摘要】：目的研究两段重复的热休克蛋白70(mHSP70)407-426序列(2mHSP70_(407-426),M_2)能否增强血管内皮生长因子(VEGF)融合蛋白疫苗抗肿瘤活性。方法以加端PCR方式,将M_2以柔性肽连接于hVEGF121突变体1的C末端,得到hVEGF121突变体2蛋白。用H22肝癌细胞构建荷瘤肝癌小鼠模型。24只小鼠按照体重随机分为3组:模型组,实验1组(hVEGF121突变体1蛋白80μg)及实验2组(hVEGF121突变体2蛋白80μg)。以酶联免疫吸附法检测免疫小鼠血清中的抗体水平,以脾淋巴细胞增殖实验检测免疫小鼠细胞免疫应答水平,以皮内肿瘤血管模型考察蛋白疫苗抗血管生成活性。结果实验1组与实验2组的瘤重分别为(1.58±0.28),(1.17±0.25)g,与实验1组比较,实验2组差异有统计学意义(P0.05)。实验1组与实验2组的小鼠免疫血清中抗-VEGF抗体分别为0.54±0.09,0.74±0.1,与实验1组比较,实验2组小鼠免疫血清中检测到更高滴度的抗-VEGF抗体,差异有统计学意义(P0.01)。实验1组与实验2组的刺激脾淋巴细胞的增殖活性分别为0.26±0.03,0.36±0.04,与实验1组比较,实验2组疫苗免疫能够更有效地刺激脾淋巴细胞的增殖,差异有统计学意义(P0.01)。结论 M_2作为分子佐剂可以有效增强VEGF融合蛋白疫苗的抗肿瘤活性。
[Abstract]:Objective to investigate whether the double repeat heat shock protein 70 (mHSP70) 407-426 (2mHSP707-426) can enhance the antitumor activity of vascular endothelial growth factor (VEGF) fusion protein vaccine. Methods Mstu2 was ligated to the C-terminal of hVEGF121 mutant 1 by adding terminal PCR. The hVEGF121 mutant 2 protein was obtained. 24 mice were randomly divided into three groups according to their body weight: model group, experimental group 1 (hVEGF121 mutant 1 protein 80 渭 g) and experimental group 2 (hVEGF121 mutant 2 protein 80 渭 g). The level of antibody in serum of immunized mice was detected by enzyme-linked immunosorbent assay (Elisa), the cellular immune response of immunized mice was detected by spleen lymphocyte proliferation test, and the anti-angiogenic activity of protein vaccine was evaluated by intradermal tumor vascular model. Results the tumor weight of group 1 and group 2 were (1.58 卤0.28), (, 1.17 卤0.25) g, which was significantly higher than that of group 1 (P0.05). The anti-VEGF antibody in the immunized serum of experimental group 1 and experimental group 2 was 0.54 卤0.09 and 0.74 卤0.1, respectively. Compared with group 1, the titer of anti-VEGF antibody was detected in the immune serum of experimental group 2. The difference was statistically significant (P0.01). The proliferative activity of splenic lymphocytes in experimental group 1 and experimental group 2 was 0.26 卤0.03 and 0.36 卤0.04, respectively. Compared with experimental group 1, vaccine immunization of experimental group 2 was more effective in stimulating the proliferation of splenic lymphocytes. The difference was statistically significant (P0.01). Conclusion MK _ 2 as a molecular adjuvant can effectively enhance the anti-tumor activity of VEGF fusion protein vaccine.
【作者单位】：滨州医学院葡萄酒学院;滨州医学院药学院方剂效应与临床评价国家中医药管理局重点实验室;淄博市沂源县人民医院;
【基金】：国家自然科学基金资助项目(81541158;8150131039) 山东省优秀中青年科学家科研奖励基金资助项目(BS2014YY051) 山东省自然科学基金资助项目(ZR2015PH002) 山东省医药卫生科技发展计划基金资助项目(2014WS0479) 山东省高校科技计划基金资助项目(J15LM51;J15LE51)
【分类号】：R392-33

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