单纯疱疹病毒1型糖蛋白D核酸疫苗的构建及初步免疫观察

目的　构建用于防治单纯疱疹的核酸疫苗并观察其免疫效果。方法　用PCR反应 ,从HSV 1F株基因组DNA中扩增出 gD蛋白的全部编码序列 ,将其插入 pcDNA 3 1 (+)的PCMV下游 ,构建HSVgD核酸疫苗 ,并用酶切分析和测序鉴定 ;通过中和试验、ELISA方法和攻击试验检测用 pLy D三次肌肉接种小鼠的免疫效果。结果　经鉴定证实了HSV gD核酸疫苗 pLy D的构建 ;实验小鼠产生了对HSV 1特异性的抗体 (抗体滴度 :ELISA法 1∶2 0 3,中和试验法 1∶37) ,并经受了HSV 1病毒的攻击 (存活率 :70 % )。结论　成功构建出了HSV gD核酸疫苗pLy D ,用其接种小鼠可以诱导产生特异性免疫反应 ,并具有良好的保护作用。

The construction and preliminary immunity of herpes simplex virus tpye 1glycoprotein D DNA vaccine

ObjectiveTo construct a DNA vaccine that ca n be used to prevent and treat the herpes simplex.MethodsThe whole encoding sequence of the glycoprotein D w as amplified from HSV1 DNA genome by polymerase chain reaction(PCR), and then w as cloned into downstream of P~{*-*)~}CMV~{**~} of pcDNA3.1( ),so as to result in the HS V1 nucleic acid vaccinepLyD, which was confirmed by the restriction endonu cle ase analysis and partial sequence. The ELISA, neutralization assay and the chall enge test were employed to evaluate immune response of the mice that were inocul ated triply with pLyD.ResultsThe construction of the HSV DNA vaccine was identifie d;~{"Z~}The inoculated mice produced specific antibodies to HSV1(ELISA titer was 1 ~{!C~}203, neutralization antibody titer was 1~{!C~}37)and survived after the HSV1 att ack.ConclusionThe HSV DNA vaccine is constructed successfully and demonstrates that it has protective effects on mice.