重组质粒pVAX1-HER2-Flt3L构建及抗肿瘤效果

发布时间：2018-04-29 22:12

本文选题：人表皮生长因子受体 + FMS相关的酪氨酸激酶配体　；参考：《生物技术》2017年06期

【摘要】：[目的]将具有免疫增效功能的Flt3L分子与HER2抗原融合表达,评价Flt3L是否能够提高抑瘤效果。[方法]将人的HER2编码区基因和小鼠Flt3L编码区基因用Furin/2A进行连接,全基因合成后,克隆至pVAX1载体,从而构建重组质粒pVAX1-HER2-Flt3L,同时构建对照质粒pVAX1-HER2和pVAX1-Flt3L。将上述质粒分别注射正常小鼠和稳定表达HER2抗原的荷瘤小鼠,评价其免疫原性和抑瘤效果。[结果]将各组重组质粒接种于Bal B/c小鼠后,抗体检测结果显示:重组融合质粒pVAX1-HER2-Flt3L组诱导产生的平均抗体滴度为2060,而单纯抗原质粒pVAX1-HER2组诱导产生的平均抗体滴度为1720;ELISPOT检测结果显示:重组融合质粒pVAX1-HER2-Flt3L组诱导产生的平均斑点数为133,而单纯抗原质粒pVAX1-HER2组诱导产生的平均抗体滴度为110;与此一致的是,在荷瘤小鼠中,该融合质粒也显示出更加显著的抑制肿瘤生长的效果,达到47%,而单纯抗原质粒仅为32%。[结论]Flt3L具有一定的免疫增效能力,能够显著提高表达HER2抗原重组质粒的抑瘤效果,深入研究其免疫保护机制具有潜在的临床意义。
[Abstract]:[objective] to evaluate whether Flt3L can improve tumor inhibition by fusion expression of Flt3L molecule with HER2 antigen. [methods] Human HER2 coding region gene and mouse Flt3L coding region gene were ligated with Furin/2A. The whole gene was synthesized and cloned into pVAX1 vector. The recombinant plasmid pVAX1-HER2-Flt3L was constructed, and the control plasmids pVAX1-HER2 and pVAX1-Flt3L were constructed. The above plasmids were injected into normal mice and tumor-bearing mice stably expressing HER2 antigen to evaluate their immunogenicity and tumor inhibition effect. [results] the recombinant plasmids were inoculated into Bal B / c mice. The results of antibody detection showed that the average titer of antibody induced by recombinant fusion plasmid pVAX1-HER2-Flt3L group was 2060, while that induced by simple antigen plasmid pVAX1-HER2 group was 1720 ELISPOT. The results showed that the recombinant fusion plasmid pVAX1-HER2-Flt3L group induced production. The average number of spots was 133, and the average antibody titer induced by pVAX1-HER2 was 110. In tumor-bearing mice, the fusion plasmid also showed a more significant inhibitory effect on tumor growth, reaching 47%, while only 32 antigenic plasmids were found in tumor-bearing mice. [conclusion] Flt3L has a certain immune synergistic ability and can significantly improve the tumor inhibition effect of recombinant plasmid expressing HER2 antigen. It is of potential clinical significance to study the immune protective mechanism of Flt3L.
【作者单位】：黑龙江中医药大学佳木斯学院;佳木斯大学附属第一医院;中国农业科学院农产品加工研究所主食加工技术研究所;
【基金】：黑龙江省自然科学基金项目(“马钱苷和莫诺苷抗恶性黑素瘤的作用机制研究”,No.D201275)
【分类号】：R730.51

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