C3d3-hCGβ融合蛋白免疫BALB/c小鼠增强抗hCG-β T_H2型体液免疫效应  

Enhance of anti-hCGβ T_H2 bias in hCGβ-C3d3 fusion protein immunized BALB/c mice

作　　者：王秀丽[1] 李大金[1] 袁敏敏[1] 

机构地区：[1]复旦大学附属妇产科研究所,上海200011

出　　处：《中华微生物学和免疫学杂志》2004年第8期666-669,共4页Chinese Journal of Microbiology and Immunology

基　　金：上海市自然科学基金项目 ( 0 0ZB14 0 5 8) ;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( 970 3 1) ;国家自然科学基金 ( 3 0 2 712 3 5 );计划生育药具国家重点实验室课题 (B2 -0 2 -1)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并实现免疫效应向TH2型体液免疫偏倚。方法　在CHO细胞中表达、纯化hCGβ、C3d3和hCGβ C3d3融合蛋白。分别用hCGβ C3d3融合蛋白、单用hCGβ、hCGβ联合C3d3和hCGβ加用弗氏完全佐剂免疫BALB c小鼠 ,共免疫 2次 ,间隔 4周。ELISA测定血清中抗hCGβ抗体滴度。末次免疫后 3周处死小鼠制备单个脾细胞悬液 ,体外用hCG刺激培养 4 8h ,ELISA检测上清中TH1型 (IFN γ、IL 2 )和TH2型 (IL 4、IL 10 )细胞因子分泌水平。结果　C3d3使hCGβ的免疫原性增强了 1995倍 ,C3d3的佐剂能力是弗氏完全佐剂的 10倍 (初次免疫 )～ 32倍 (再次免疫 )。借助C3d3分子佐剂 ,hCGβ C3d3融合蛋白可产生很明显的TH2型体液免疫优势效应。结论　分子佐剂C3d3可以大幅增强hCGβ避孕疫苗的免疫原性 ,使免疫效应向TH2型体液免疫偏倚。ObjectiveTo enhance the immunogenicity and T_H2-biased humoral immunity of hCGβ contraceptive vaccine via the molecular adjuvant C3d3. MethodshCGβ,C3d3 and hCGβ-C3d3 fusion protein were gained and purified. BALB/c mice were inoculated with hCGβ-C3d3 fusion protein,hCGβ,hCGβ plus C3d3 and hCGβ plus CFA/IFA respectively,followed by a boost at the day 28. The antibody titer of serum was determined by indirect enzyme-linked immunosorbent assay (ELISA). At the third week after the last inoculation,the spleen cells of the immunized mice were prepared and stimulated by hCG for 48 h. Then the cytokines of T_H1(IFN-γ,IL-2)and T_H2(IL-4,IL-10) in the supernatant were determined by ELISA. ResultsThe hCGβ-C3d3 fusion protein was 1995-fold and 10-32-fold more immunogenic than hCGβ alone and hCGβ plus CFA/IFA respectively. ConclusionMolecular adjuvant C3d3 greatly enhances the immunogenicity and T_H2 bias of hCGβ protein vaccine and this approach may be a novel approach to enhance the immunogenicity of hCGβ contraceptive vaccine

关 键 词：C3d3-hCGβ融合蛋白 免疫学 BALB/C 小鼠 抗hCG-βTH2型 体液免疫效应 免疫原性 

分 类 号：R392[医药卫生—免疫学]