肝片吸虫DNA疫苗的构建及其免疫效应的初步研究

用ＥcorRI酶切含肝片吸虫保护性抗原基因ＦＨ３的重组质粒pUC18/FH3,回收ＦＨ３（－１．０Ｋb)片段克隆到真核表达载体pBlueCMV上，酶切筛选顺向插入重组子pBlueCMV-FH3，即得到一种肝片吸虫ＤＮＡ疫苗，制备纯化该疫苗并免疫小鼠，小鼠肌肉细胞中有一定的ＦＨ３抗原表达；用ＥＬＩＳＡ检测抗血清表明，注射疫苗的小鼠均伴随产生一定量的特异性抗体，用肝片吸早囊蚴攻击（２０囊蚴／鼠）小鼠，初步显示有一定的减虫率。

CONSTRUCTION A DNA VACCINE OF FASCIOLA.HEPATICA AND THE PRIMARY STUDY ON ITS IMMUNE RESPONSES

Proctective antigen gene FH3 of F. hepatica was cut out from recombinant plasmid pUC18/FH3 using EcoRI and then cloned into a eukaryotic expression vector pBlueCMV,and a F. hepatica DNA vaccine pBlueCMV-FH3 was constructed. After prelation and purification, the DNA vaccine was transferred into Kunming mice in the amount of 100ug/mouse through intramuscular inoculation. Muscle tissues of some mice were cut off to test the expression of FH3 antigen, and the result showed that FH3 antigen was indeed expressed. The humoral immune response results showed that this vaccine could induce specific antibodies,Furthermore, the titers of mice immuned 3 times were much higher than those of immuned single time. After immuned 3 times, some mice were challenged with cysticercus (20 cysticercus per mouse) of F. hepatica,and the immuno-protection rate was 41%.