CEA迷你肽表位基因疫苗诱导小鼠抗肿瘤免疫的研究

发布时间：2018-01-31 17:14

本文关键词： 癌胚抗原迷你基因基因疫苗肿瘤生物治疗　出处：《中国免疫学杂志》2017年03期 　论文类型：期刊论文

【摘要】：目的:利用含有CEA_(625-667)基因的单倍体疫苗pc DNA-CEA_(625-667)及三串联体的DNA疫苗pc DNA-tri CEA_(625-667)免疫小鼠后,观察其诱导的抗肿瘤免疫效应。方法:采用4~6周纯系BALB/c小鼠,肌肉注射法分pc-DNA3.0、pc DNACEA_(625-667)、pc DNA-tri CEA_(625-667)三组免疫小鼠,对其激发的机体特异性及非特异性免疫反应进行研究。流式细胞术检测免疫小鼠脾细胞的T细胞亚群和CD4~+/CD8~+比值;3H-TdR掺入法检测免疫小鼠的特异性淋巴细胞增殖;Western blot杂交及ELISA法检测免疫小鼠血清中的CEA特异性抗体;ELISA法检测免疫动物脾细胞体外诱导IFN-γ、IL-4、GM-CSF的分泌水平。结果:实验组与对照组的CD4~+/CD8~+比值差异无统计学意义;经过基因疫苗免疫的小鼠与天然小鼠相比,其脾细胞在体外与短肽共孵育之后,会在更短的时间内出现更明显的细胞增殖;免疫了CEA迷你基因串联体肿瘤疫苗的小鼠血清中可以产生低滴度的抗体,提示HTL的活化;免疫小鼠脾细胞上清中IFN-γ的含量明显高于对照组,而pc-DNA3.0,pc DNA-CEA_(625-667),pc DNA-triCEA_(625-667)各组IL-4的含量均很低,差异无统计学意义;迷你基因三倍体疫苗所引发的增殖效应以及释放细胞因子的水平均高于迷你基因一倍体,说明我们所采用将目的基因多倍串联的抗原改造的方式起到了增强免疫效应的作用。结论:CEA迷你肽表位基因单倍体及三倍体疫苗均不能显著改变动物体内CD4/CD8比值,但均能诱导HTL活化,使被免疫机体T细胞趋向于Th1效应且三倍体疫苗的免疫原性强于单倍体疫苗。
[Abstract]:Objective: to make use of the haploid vaccine PC DNA-CEA virus containing the gene CEAX 625-667). Mice were immunized with DNA vaccine PC DNA-tri CEA (625-667). Methods: BALB/c mice were divided into two groups by intramuscular injection of pc-DNA 3.0 and pcDNACEA (625-667). The mice were immunized with PC DNA-tri. The specific and nonspecific immune responses stimulated by the immunized mice were studied. The T cell subsets and CD4 / / CD8 ~ ratio of spleen cells were detected by flow cytometry. The specific lymphocyte proliferation of immunized mice was detected by 3H-TdR incorporation. Western blot hybridization and ELISA assay were used to detect the CEA specific antibody in the serum of immunized mice. ELISA assay was used to detect IFN- 纬 IL-4 induced by splenocytes of immunized animals in vitro. Results: there was no significant difference in the ratio of CD4 to / / CD8 ~ between the experimental group and the control group. Compared with the natural mice, the spleen cells of the mice immunized with the gene vaccine could proliferate more obviously in a shorter time after the spleen cells were incubated with the short peptide in vitro. Mice immunized with CEA mini gene tandem tumor vaccine could produce low titer antibody, indicating the activation of HTL. The content of IFN- 纬 in the supernatant of spleen cells of immunized mice was significantly higher than that of the control group, while pc-DNA 3.0 pc-DNA-CEA was found in the spleen cell supernatant of immunized mice. The content of IL-4 in each group was very low, and the difference was not statistically significant. The proliferative effect and the release level of cytokines induced by the triploid vaccine were higher than those of the triploid gene. The results showed that the antigenic modification of the multiplex series of the target gene played an important role in enhancing the immune response. Conclusion:. Neither haploid nor triploid vaccine of CEA mini peptide epitope gene could significantly change the CD4/CD8 ratio in animals. All of them could induce the activation of HTL, and the immunogenicity of triploid vaccine was stronger than that of haploid vaccine.
【作者单位】：吉林省人民医院医学诊治实验中心;吉林大学第一医院呼吸内科;
【基金】：吉林省科技厅国际合作项目(20140414043GH);吉林省科技厅中青年科技创新领军人才及团队项目(20140519018JH);吉林省科技厅重点实验室项目(20122113)资助吉林省人社厅省人才开发资金(2016年度)
【分类号】：R392
【正文快照】： 1本文受吉林省科技厅国际合作项目(20140414043GH)、吉林省科技厅中青年科技创新领军人才及团队项目(20140519018JH)、吉林省人社厅省人才开发资金(2016年度)和吉林省科技厅重点实验室项目(20122113)资助。2吉林大学第一医院呼吸内科,长春130021。癌胚抗原(Carcinoembryonic a

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