【摘要】 目的　构建人巨细胞病毒 (HCMV)pp6 5基因表达载体并初步评价其作为核酸疫苗的体内免疫效果。方法　利用分子克隆方法构建HCMVpp6 5表达载体pcDNA3.pp6 5 ;对其活性进行体外鉴定后 ,以肌肉和皮下 2种途径、3种剂量组免疫小鼠 ;测量免疫小鼠的体重、体温等一般状况 ,利用MTT方法测定免疫小鼠的T细胞增殖活性、NK活性 ,利用ELISA方法测定小鼠的抗HCMVIgM、IgG抗体 ,免疫小鼠血清的病毒中和实验。结果　成功构建HCMVpp6 5表达载体pcDNA3.pp6 5并免疫小鼠后 ,小鼠体重、体温等一般状况未见明显变化 ,T细胞增殖活性增强 ,NK细胞活性增强 ,HCMVIgM、IgG抗体增高 ,且有时间、剂量依赖性 ,免疫小鼠血清对HCMV有部分中和作用。结论　pcDNA3.pp6 5表达载体可引起机体特异性细胞免疫和体液免疫 ,可作为一种有一定应用前景的核酸疫苗进一步深入研究。更多还原

【Abstract】 Objective To construct a HCMV pp65 expression vector and to test in vivo its immunological effects. Methods Constructed HCMV pp65 expression vector pcDNA3.pp65 by molecular biology methods. Detected the biological activity of pcDNA3.pp65 in vitro . BALB/c mice were immunized by pcDNA3.pp65 by i.m and hypodermic injection. The weight and temperature were measured after immunization. T cell proliferation activity, NK activity and serum neutralization for HCMV were tested. Serum IgM and IgG levels of HCMV were detected by ELISA method. Results The expression vector pcDNA3.pp65 was successfully constructed. The T cell proliferation activity, NK cell activity, HCMV IgM and IgG antibodies were increased with the time after immunization and dose. It appeared that the immunized serum could neutralize HCMV virulence, but there was no significant difference between the control and the experimental mice. Conclusion HCMV DNA vaccine of pcDNA3.pp65 could induce cellular and humoral immunological reaction.更多还原