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(P<0.05)。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(P<0.01)。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(P<0.01)。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。
Objective To investigate whether 2 repeats of mycobacterial microbial hot shock protein 70（ HSP70） 407-426（ 2mHSP70407-426,M_2） could act as an effective molecular adjuvant to enhance the anti-tumor efficiency of vascular endothelial growth factor（ VEGF） fusion protein vaccine. Methods The gene encoding two tandem repeat sequences of HSP70407-426 was introduced to the terminus of the constructed hVEGF121 mutant 1 gene to prepare a recombined hVEGF121 mutant 2protein vaccine by PCR technique. The anti-tumor efficacy of hVEGF121 mutant 2 protein vaccine was investigated using a H22 liver cancer subcutaneous tumor model. The 24 mice were randomly divided into three groups： model group,experiment 1 group（ 80 μg hVEGF121 mutant 1 protein） and experiment 2 group（ 80 μg hVEGF121 mutant 2protein）. The humoral and cellular immune responses were detected by ELISA and splenic lymphocyte proliferation assay. The anti-angiogenesis effect was evaluated by an intradermal tumor model. Results Thetumor weight of the experimental 1 group and the experimental 2 group were（ 1. 17 ± 0. 25） g and（ 1. 58 ± 0. 28） g.Compared with the experimental 1 group,the tumor weight of tumor bearing mice in the experimental 2 group was lower significantly（ P〈0. 05）. The anti-VEGF antibodies of the experimental 1 group and experimental 2 group was 0. 54 ± 0. 09,0. 74 ± 0. 1. Compared with experimental 1 group,higher titers of anti-VEGF antibody were detected in immune serum of the experimental 2 group with statistically significant（ P〈0. 05）. The proliferative activity of spleen lymphocytes in the experimental 1 group and the experimental 2 group was 0. 26 ± 0. 03,0. 36 ± 0. 04. Compared with the experimental 1 group,the vaccine immunization in experimental 2 group can stimulate the proliferation of spleen lymphocytes more effectively with statistically significant（ P〈0. 05）. Conclusion M_2 could act as an effective adjuvant to help VEGF fusion protein vaccine to elicit a strong anti-tumor effi
出处
《中国临床药理学杂志》
CAS
CSCD
北大核心
2017年第13期1225-1228,共4页
The Chinese Journal of Clinical Pharmacology
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)
关键词
血管内皮生长因子
热休克蛋白70
分子佐剂
蛋白疫苗
肝癌
vascular endothelial growth factor
microbial heat shock protein 70
molecular adjuvant
recombined protein vaccine
hepatocellular carcinoma
