重组新城疫病毒疫苗rl-RVG对A_(549)荷瘤小鼠瘤体生长的抑制作用

The tumor growth influence of recombinant NDV(rl-RVG) on lung adenocarcinoma A_(549) transfected in vivo

目的:探讨稳定表达狂犬病毒糖蛋白的重组新城疫病毒(NDV)疫苗(rl-RVG)对人肺腺癌A549细胞荷瘤鼠的生长抑制及其可能的免疫机制。方法:建立A_(549)荷瘤鼠模型并随机分为rlRVG组、NDV组和对照组,每组10只,分别于瘤体注射rl-RVG,NDV和PBS,每周2次,共3周。测量三组瘤体体积;PCR法测NDV及狂犬病毒糖蛋白(RVG)的表达,蛋白印迹法检测皮下瘤体组织RVG、NDV蛋白表达;流式细胞术检测荷瘤小鼠脾脏自然杀伤细胞(NK)细胞数。结果:与对照组相比,rl-RVG组及NDV组荷瘤鼠瘤体增长明显缓慢;NDV在rl-RVG组和NDV组中均有表达,而RVG在rl-RVG组中表达,对照组未见二者表达;rl-RVG组及NDV组瘤体NK细胞数明显增多,且rl-RVG组明显高于NDV组。结论:rl-RVG成功转染至A549荷瘤鼠,对其瘤体生长有着明显的抑制作用,可能与rl-RVG激活机体的免疫反应相关。

Objective:To explore the effects of recombinant avirulent newcastle disease virus(NDV)LaSota strain expressing the rabies virus glycoprotein(rl-RVG)on the proliferation of A549lung adenocarcinoma tumor-bearing mice and its immune mechanism.Methods:A tumor model of human A549into nude mice was created and divided randomly into rl-RVG group,NDV group and control group.The mice in each group received rl-RVG,NDV and PBS injection,respectively,twice a week,for3weeks.The growth of tumor were recorded,and the expression of NDV and rabies virus glycoprotein,RVG were detected by PCR and Western blot.The number of NK(CD3-CD49+)cells were detected by flow cytometry.Results:Compared with control group,the growth of tumor in rl-RVG group and NDV group were inhibited more effectively.NDV was expressed in rl-RVG group and NDV group,RVG was expressed in rl-RVG group,while both two were not expressed in control group.The number of NK cells was significantly higher in rl-RVG and NDV groups than in the PBS group.All the phenomenons were more obvious in rl-RVG group than that in NDV group.Conclusion:Recombinant rl-RVG transfected successfully,and effectively inhibited the growth of A549lung adenocarcinoma by mechanism which inducing regulating cell immune response.