CAJ | 학술논문

为了研究基因疫苗对绵羊的免疫保护效果,本研究构建了捻转血矛线虫(H.contortus)Hc38基因DNA疫苗.将H.contortus Hc38基因保守结构域克隆到真核表达载体pcDNA3.1中,免疫鼠8d后用RT-PCR检测到该疫苗在鼠肌肉组织中进行了转录.将纯化的DNA疫苗免疫绵羊后,用western blot和ELISA方法检测疫苗在绵羊体内的翻译和诱导IgG的产生.二免后2周用10 000条H.contortus第3期幼虫攻击实验动物,检测绵羊粪便虫卵排出、成虫数量等免疫保护性指标.该H.contortus Hc38 DNA疫苗与对照组比较,免疫组绵羊排出虫卵减少66.6%、成虫减少33.1%.特别值得注意的是免疫组的静脉注射方式产生抗体最高,相应羊的虫卵数和成虫数低.本实验证明Hc38基因DNA疫苗对绵羊虫卵及成虫发育具有明显的抑制作用.
위료연구기인역묘대면양적면역보호효과,본연구구건료념전혈모선충(H.contortus)Hc38기인DNA역묘.장H.contortus Hc38기인보수결구역극륭도진핵표체재체pcDNA3.1중,면역서8d후용RT-PCR검측도해역묘재서기육조직중진행료전록.장순화적DNA역묘면역면양후,용western blot화ELISA방법검측역묘재면양체내적번역화유도IgG적산생.이면후2주용10 000조H.contortus제3기유충공격실험동물,검측면양분편충란배출、성충수량등면역보호성지표.해H.contortus Hc38 DNA역묘여대조조비교,면역조면양배출충란감소66.6%、성충감소33.1%.특별치득주의적시면역조적정맥주사방식산생항체최고,상응양적충란수화성충수저.본실험증명Hc38기인DNA역묘대면양충란급성충발육구유명현적억제작용.
To study protective effects of Hc38 DNA vaccines against Haemonchus contortus in sheep, we constructed DNA vaccines containing conserved Hc38 protein domain. The Hc38 gene was inserted into pcDNA3.1 and administered to mice. Transcriptions of the vaccines were confirmed by RT-PCR in mouse muscles. Sheep were immunized with DNA vaccine and antibody (IgG) production was analyzed by western blot and ELISA. After 2 weeks, all sheep were challenged by 10000 H. Contortus L3 strain. The results showed that EPG (eggs per gram feces) and worm burdens in immunized sheep reduced by 66.6 % and 33.1%, compared to the control group. Vaccination by intravenous injection induced highest antibody level and resulted in lowest EPG and worm burdens in sheep.