人巨细胞病毒gB基因真核表达载体在小鼠中的免疫效果

Immune Effect of DNA of Eukaryotic Expression Vector of Human Cytomegalovirus (HCMV) Glycoprotein B in Mice

目的观察人巨细胞病毒(HCMV)gB基因真核表达载体在小鼠中的免疫效果.方法大量提取目的质粒,分3个剂量组进行多点肌肉注射免疫小鼠,利用MTT法检测免疫小鼠的T细胞增殖活性,ELISA法检测小鼠血清中HCMV特异性抗体,中和试验检测小鼠血清中和抗体.结果 ELISA检测结果表明,与空白对照、空载体对照组相比,3个剂量组pcDNA3.1/gB质粒免疫后小鼠血浆中抗HCMV IgG抗体水平均明显增高(P＜0.05),中和抗体水平为1:20～1:40,而对照组血清中无中和抗体存在.淋巴细胞增殖指数免疫小鼠与正常小鼠差异无显著意义.结论已构建的HCMVgB基因真核表达载体可以诱导小鼠产生明显的体液免疫,为核酸疫苗研究奠定了基础.

Objective To observe the immune effect of DNA of eukaryotic expression vector of human cytomegalovirus（HCMV） glycoprotein B（gB） in mice.Methods Three groups of BALB/c mice were injected i.m. with the extracted plasmid pcDNA3.1/gB at three dosages respectively. The T lymphocyte proliferation activity, serum HCMV specific antibody and serum neutralizing antibody of the mice were detected by MTT method, ELISA and neutralization test respectively. Results ELISA showed that, compared with those of mice in blank control and empty vector control groups,all the serum anti-HCMV IgG levels of mice immunized with pcDNA3.1/gB at three dosages improved significantly （ P 〈 0.05）. The serum neutralizing antibody titers of the immunized mice were 1 ： 20-1 ： 40, however, no neutralizing antibody were detected in the sera of mice in two control groups. T lymphocyte proliferation test showed no significant difference between the stimulating indexes of immunized and normal mice, Conclusion The constructed eukaryotic expression vector of HCMV gB gene induced significant humoral immunity. It laid a foundation of study on DNA vaccine.