HCV高变区1模拟表位串联基因诱导免疫应答的实验研究

To study the immune response induced by tandem gene of mimic epitopes in hypervariable region 1 of HCV

目的:研究丙型肝炎病毒(HCV)高变区1(HVR1)模拟表位的串联基因免疫后在小鼠体内引起的免疫反应特点。方法:10只Balb/C小鼠随机分为2组,每组5只,试验组给予pCDNA3.1-target gene质粒免疫,对照组给予pCDNA3.1质粒免疫。采用淋巴细胞增殖实验检测脾细胞增殖情况,并检测小鼠脾细胞培养上清中的细胞因子(IL-4、10,IFN-,γTNF-α)水平。结果:肽1、肽5、肽6、肽7刺激基因免疫小鼠脾细胞后,小鼠脾细胞平均增殖指数(PI)分别为71.45±3.1、12.64±2.5、10.05±1.9和31.20±2.4。培养上清中干扰素-γ,肿瘤坏死因子-α,白介素-4,白介素-10都有不同程度的提高。结论:基因免疫使Th1和Th2类细胞因子水平都有所升高,诱导产生了表位特异性的细胞免疫反应。

Objective： to study the characteristic of immune response induced by tandem gene of mimic epitopes in hypervariable region 1 of HCV. Methods：10 Balb/C mice were divided into two groups. Group one was immuned with plasmid pCDNA3.1-target gene and group two was immuned with plasmid pCDNA3.1. Splenocyte proliferation was tested by 3 H-TdR proliferation assay and the level of cytokines in culture supematant was tested by ELISA. Results： splenocyte of immuned mice were proliferate greatly when stimulated by HVRI antigen synthetic peptides. The PI were 71.45± 3, 1, 12, 64 ±2.5, 10, 05±1.9, and 31.20±2.4 respectively after stimulate by peptide 1,2, 3 or 4. IL-4, IL-10, IFN-γ and TNF-α also increased in culture supernant, Conclusion：the level of Thl and Th2 cytokines were increased after immunation and epitope-specific cellular immune response was induced.