摘要
将猪细小病毒VP2基因克隆至pCI-neo真核表达载体中,构建了pCIneo-VP2重组质粒,转染至PK-15细胞中,利用免疫荧光方法检测在体外表达情况;并以小鼠为动物模型,将pCIneo-VP2、pCI-neo重组质粒、猪细小病毒活疫苗和对照组通过肌肉注射进行免疫,检测免疫小鼠的淋巴细胞转化功能,特异性CTL杀伤活性和血清抗体滴度。结果显示,pCIneo-VP2在体外能够诱导PK-15细胞表达VP2蛋白,小鼠注射pCIneo-VP2质粒1周后能够诱导机体产生抗体,4周时达到峰值,与活疫苗对照组产生的抗体滴度、诱导T淋巴细胞增殖和诱导强的细胞毒性基本一致。试验表明,构建的pCIneo-VP2能够有效诱导机体产生体液免疫和细胞免疫,为研制出高效、新型猪细小病毒疫苗提供了科学依据和试验依据。
To construct gene vaccine of Porcine parvovirus （PPV） and to investigate its immune response in mice, the recombinant eukaryotic expression plasmid of pCIneo-VP2 was constructed and transfected into PK-15 cells by lipofectamine, the expressed product was detected by immunofluorescence assay. To study the immune responses of DNA vaccine in vivo, the recombinant plasmid pCIneo-VP2, the control pCI-neo and the PPV live vaccine were immunized by intramuscular injection in mice. Anti-PPV antibodies were measured by ELISA. lymphocyte proliferation activity was detected using MTY method, and the specific CTL were assayed too. The results show that the immunized mice produced PPV antibody after one week, and reached to the highest level after four weeks. Compared with the control group, the pCIneo-VP2 immunized group produced significant differences in the antibody titers, the lymphocyte proliferation activity and the specific killing activities of CTL. The pCIneo-VP2 induced humora and cellular immune responses similarly to that the live vaccine induced. These results manifested that the PPV DNA vaccine successfully induced humoral and cellular immune response in mice.
出处
《中国生物工程杂志》
CAS
CSCD
北大核心
2006年第12期63-67,共5页
China Biotechnology
基金
国家"十五"食品安全重大攻关专项(2001BA804A30-11)
河南省"重点"科技攻关项目(0123011500)
