分子佐剂C3d增强草原兔尾鼠卵透明带3DNA疫苗的免疫应答  

ENHANCING THE INDUCED IMMUNE RESPONSES OF LAGURUS LAGURUS ZONA PELLUCIDA 3 DNA VACCINE BY MOLECULAR ADJUVANT C3d

作　　者：王焱冰[1] 李轶杰[1] 张富春[1] 

机构地区：[1]新疆大学生命科学与技术学院新疆生物资源基因工程重点实验室,乌鲁木齐830046

出　　处：《分子细胞生物学报》2006年第4期297-303,共7页Journal of Molecular Cell Biology

基　　金：国家自然科学基金资助项目(No.30360062)

摘　　要：为增强免疫不育的效果,探讨分子佐剂C3d对草原兔尾鼠透明带3(LaguruslagurusZonaPellucida3,LZP3)基因免疫的调节作用。构建了含3个拷贝C3d的lzp3基因真核细胞表达质粒pcDNA3-LZP3-C3d3(pcD-LC),以不含C3d的质粒pcDNA3-LZP3(pcD-L)为对照,体外转染HeLa细胞,RT-PCR和Westernblot分别检测到lzp3mRNA和蛋白的表达。对NIH雌性小鼠肌肉进行基因免疫注射,ELISA结果表明pcD-LC免疫诱导的特异性抗LZP3-IgG水平明显高于pcD-L对照组(P<0.05),且有长期免疫应答效应;MTT结果显示pcD-LC能够诱导淋巴细胞增殖活性的提高。抗生育实验表明pcD-LC免疫组显著降低窝产仔数(P<0.05),且卵巢切片正常。研究结果证实C3d能够增强lzp3DNA疫苗的特异性免疫应答并达到抗生育效果,为DNA疫苗控制草原兔尾鼠的深入研究提供了基础。To investigate the enhancement immunization induced by molecular adjuvant of Lagurus lagurus zona pellucida 3 gene （lzp3） C3d in comparison with plasmid pcDNA3-LZP3 （pcD-L） in non-C3d fused form, the eukaryotic expression vector pcDNA3-LZP3-C3d3（pcD-LC） including lzp3 and three copies of C3d in fusion form was constructed. In vitro,RT-PCR and Western blot showed that lzp3 mRNA and protein were expressed in HeLa cells transfected with pcD-L and pcD-LC, respectively. In vivo,the result of ELISA showed that NIH female mice vaccined with pcD-LC elicited higher titer, longer lasting anti-LZP3 antibody than non-C3d fused form pcD-L （P〈0.05） by intramuscular gene immunization. MTT analysis indicated that the lymphocytes proliferation response of the pcD-LC primed mice was significantly stronger than that of the blank plasmid injected mice. Antifertility experiment showed that pcD-LC had enhanced effect on blocking fertility of mice （P〈0.05） with no disruption of follicular development. These results proved that C3d enhanced immune responses induced by lzp3 vaccine and antifertility effect,and provided the basis for the future study of the Lagurus lagurus reproductive controlling.

关 键 词：草原兔尾鼠卵透明带3 DNA 疫苗 分子佐剂C3d 免疫应答 抗生育 

分 类 号：S812.6[农业科学—草业科学]