分子佐剂C3d3增强hCGβ蛋白疫苗的体液免疫效应  

Enhancement of antibodies to the hCGβ contraceptive vaccine by using the molecular adjuvant C3d3

作　　者：王秀丽[1] 李大金[1] 袁敏敏[1] 王明雁[1] 朱影[1] 孟毅[1] 

机构地区：[1]复旦大学附属妇产科研究所,上海200011

出　　处：《中国免疫学杂志》2005年第9期652-655,658,共5页Chinese Journal of Immunology

基　　金：上海市自然科学基金(00ZB14058);教育部博士学科点专项科研基金(9941);卫生部优秀青年人才专项科研基金(97031);国家自然科学基金(30271235);计划生育药具国家重点实验室课题(B2021)资助

摘　　要：目的:通过分子佐剂C3d3增强hCGβ避孕疫苗的免疫原性。方法:采用分子生物学技术以phCMV1为载体分别构建分泌型、带有6个组氨酸纯化标签的真核表达载体phCMV16HishCGβC3d3和phCMV16HishCGβ,在CHO细胞中获得重组蛋白的稳定、高效表达并用镍柱和凝胶过滤层析对其进行分离、纯化。分别用hCGβC3d3融合蛋白、hCGβ联合C3d3和单用hCGβ对BALBc与C57BL6小鼠进行免疫,共免疫两次,间隔4周,ELISA测定血清中的抗体滴度,比较各组的免疫效果。结果:无论是BALBc小鼠还是C57BL6小鼠,hCGβC3d3融合蛋白诱导产生的抗hCGβ抗体的出现时间都明显早于hCGβ与C3d3联合免疫和hCGβ单独免疫组。在BALBc小鼠,初次和再次免疫结果显示,C3d3使hCGβ的免疫原性增强了1995倍。C57BL6小鼠所显示出的体液免疫反应强度虽不如BALBc小鼠,但C3d3也使hCGβ的免疫原性增强了1259倍。结论:通过分子佐剂C3d3可以大幅地提高hCGβ避孕疫苗的免疫原性,在个性差异很大的不同品系小鼠,C3d3均能增强机体对hCGβ的体液免疫应答能力。Objective：To enhanee the immunogenity of hCGβ contraceptive vaccine by using the molecular adjuvant C3d3. Methods： phCMV1-6His-hCGβ and phCMV1-6His-hCGβ-C3d3 expression vectors,based on the phCMV1 ,were constructed by way of molecular cloning. The recombinant 6His-hCGβ and 6His-hCGβ-C3d3 fusion proteins were gained by stable expression in CHO cells. The recombinant proteins of interest were purified using the immobilized metal-ion affinity chromatography（IMAC） under native conditions and Sephadex G-150 column gel filtration chromatography. BLAB/c and C57BL/6 mice were inoculated with hCGβ-C3d3 fusion protein, hCGβ plus C3d3 and hCGβ respectively, followed by a boost at 28 days. Antibody titers of serum were determined by enzyme-linked immunosorbent assay（ELISA）. Results： We have successfully constructed the eukaryotie expression vectors of phCMV1-6His-hCGβ and phCMV1-6His-hCGβ-C3d3 and gained adequately the correspondent recombinant proteins from the transfected CHO cell culture supematant. The hCGβ-C3d3 fusion protein achieved an earlier immune response than hCGβ plus C3d3 or hCGβ alone both on BALB/c and C57BL/6 mice. hCGβ-C3d3 fusion protein was 1 995-fold and 1 259-fold more immunogenic respectively in BALB/c and C57BL/6 mice than hCGβ alone. Conclusion： Molecular adjuvant C3d3 could enhances the antibody responses to hCGβ contraceptive vaccine compared to non-C3d fused forms of hCGβ, and this approach might be one way to overcome the poor immunogenicity of hCGβ contraceptive vaccine.

关 键 词：分子佐剂 C3D3 HCGΒ 避孕疫苗 体液免疫 

分 类 号：R169.41[医药卫生—公共卫生与预防医学]