分子佐剂C3d增强hCGβ基因免疫体液免疫效应

Molecular adjuvant C3d can improve humoral immunity of hCGβ DNA immunization

目的 :该实验室前期工作已成功构建真核表达质粒pcDNA3 hCGβ、pcDNA3 hCGβ C3d3且证明其能在真核表达系统中有效表达。通过动物DNA免疫 ,以期证实分子佐剂C3d增强免疫避孕疫苗免疫原性。方法 :抽提与纯化pcDNA3、pcD NA3 hCGβ、pcDNA3 hCGβ C3d3质粒 ,进行动物DNA免疫。免疫剂量分别为 5、10、2 0pmol,共免疫 2次 ,每次间隔 3周。末次免疫后 6周采血 ,用间接ELISA分析实验动物外周血抗hCGβ抗体效价。 结果 :C3d分子佐剂能明显提高抗hCGβ抗体滴度 ;且其作用呈剂量依赖性。结论 :分子佐剂确实能增强hCGβ免疫原性 ;此结果将有助于免疫避孕疫苗的技术进步及实际应用。

Objective:In the past study, a plasmid of pcDNA3 hCGβ C3d3 had been constructed. It was suggested that the fusing protein be expressed by the pcDNA3 hCGβ C3d3 plasmid both in the transient expression system in COS 7 cells, and the stable expression system in CHO cells. In the present research, it would be testified that the C3d molecular adjuvant could improve the immunogenicity of the hCGβ DNA immunization or not. Methods:BALB/C mice of 6 weeks old were immunized intramuscularly two times at interval of 3 weeks by the plasmid pcDNA3, pcDNA3 hCGβ, pcDNA3 hCGβ C3d3 at dosage of 5, 10, 20 pmol, respectively. The anti hCGβ antibody titers were determined by indirect ELISA in 6 weeks of last immunization. Results:The result showed that the C3d molecular adjuvant could increase significantly the titer of anti hCGβ antibody in a dose dependent manner after DNA immunization.Conclusion:The C3d molecular adjuvant does improve the humoral immunity of the hCGβ DNA immunization, which would be helpful for technical progress and clinical trial of the contraceptive vaccine. [