Reduction of fertility in male mice immunised with pSG.SS.C3d3.YL.Bin1b recombinant vaccine

Abstract

Objectives To study the anti-fertility effect of a DNA vaccine using Bin1b as the target antigen in male mice.

Methods A novel recombinant eukaryotic vector containing a fusion gene sequence of mouse Bin1b in tandem with three copies of C3d fragment (C3d3) was used to construct pSG.SS.C3d3.YL.Bin1b. The correct expression of the Bin1b-C3d3 protein was confirmed in transfected HEK293 cells by indirect immunofluorescence and western blot analysis. The fertility of immunised mice was determined by a mating experiment and sperm motility test. Anti-Bin1b antibody titres in sera were examined by ELISA assays. Binding activity of C3d3 fragment of the fusion protein was verified in C3d receptor-expressing Raji cells and flow cytometric analysis.

Results Immunisation of pSG.SS.C3d3.YL.Bin1b recombinant DNA vaccine significantly decreased sperm motility and compromised fertility in male mice. ELISA results showed that the titres of anti-Bin1b IgG in sera of immunised mice increased markedly with the immunisation process. Further, the anti-fertility effect of pSG.SS.C3d3.YL.Bin1b was significantly better than that of pSG.SS.YL.Bin1b DNA vaccine and generated higher titres of anti-Bin1b antibody.

Conclusions Our results show that recombinant DNA vaccine targeting Bin1b can markedly reduce fertility in male mice, providing an alternative approach for birth control.

Chinese Abstract

摘要

目的研究以Bin1b作为雄性小鼠靶抗原的DNA疫苗的抗生育作用。

方法使用包含小鼠Bin1b与三拷贝C3d片段（C3d3）融合基因序列的新型重组真核载体构建pSG.SS.C3d3.YL.Bin1b。间接免疫荧光和蛋白印迹分析证实Bin1b-C3d3蛋白在转染HEK293细胞中正确表达。交配试验和精子运动试验来确定免疫小鼠的生育能力。ELISA 检测血清中的抗Bin1b抗体滴度。C3d受体表达Raji细胞和流式细胞分析验证融合蛋白中C3d3片段的结合活性。

结果免疫接种pSG.SS.C3d3.YL.Bin1b重组NDZ疫苗显著降低雄性小鼠的精子运动力，损害其生育能力。ELISA 结果表明，免疫过程中雄性小鼠抗Bin1b抗体IgG的血清滴度显著降低。而且，pSG.SS.C3d3.YL.Bin1b产生较高的抗Bin1b抗体滴度，其抗生育效应明显优于pSG.SS.YL.Bin1b DNA疫苗的抗生育效应。

结论我们的研究结果表明，以Bin1b为靶抗原的重组DNA疫苗能够显著降低雄性小鼠的生育能力，为生育控制提供了一种新的方法。

关键词

抗生育能力，Bin1b，C3d，重组载体疫苗

Keywords:

• Anti-fertility
• Bin1b
• C3d
• Recombinant vector vaccine

Declaration of interest: The authors report no conflicts of interest. The authors alone are responsible for the content and writing of the paper.