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The preparation and application of N-terminal 57 amino acid protein of the follicle-stimulating hormone receptor as a candidate male contraceptive vaccine

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Asian Journal of Andrology 2014年第4期16卷 623-630页

作者： Cheng Xu Ying-Chun Li Hua Yang Yan Long Min-Jian Chen Yu-Feng Qin Yan-Kai Xia Ling Song Ai-Hua Gu Xin-Ru Wang State Key Laboratory of Reproductive Medicine Institute of Toxicology Nanjing China Key Laboratory of Modern Toxicology of Ministry of Education School of Public Health Nanjing China Department of Physiology Nanjing Medical University Nanjing China Yangz.hou Center for Disease Control and Prevention Yangzhou China Department of Pharmacology China Pharmaceutical University Nanjing China.

Follicle-stimulating hormone receptor （FSHR）, which is expressed only on Sertoli cells and plays a key role in spermatogenesis, has been paid attention for its potential in male contraception vaccine research and development. This study introduces a method for the preparation and purification of human FSHR 57-amino acid protein （FSHR-57aa） as well as determination of its immunogenicity and antifertility effect. A recombinant pET-28a（＋）-FSHR-57aa plasmid was constructed and expressed in Escherichia coil strain BL21 StarTM （DE3） and the FSHR-57aa protein was separated and collected by cutting the gel and recovering activity by efficient refolding dialysis. The protein was identified by Western blot and high-performance liquid chromatography analysis with a band of nearly 7 kDa and a purity of 97.4%. Male monkeys were immunized with rhFSHR-57aa protein and a gradual rising of specific serum IgG antibody was found which reached a plateau on day 112 （16 weeks） after the first immunization. After mating of one male with three female monkeys, the pregnancy rate of those mated with males immunized against FSHR-57aa was significantly decreased while the serum hormone levels of testosterone and estradiol were not disturbed in the control or the FSHR-57aa groups. By evaluating pathological changes in testicular histology, we found that the blood-testis barrier remained intact, in spite of some small damage to Sertoli cells. In conclusion, our study demonstrates that the rhFSHR-57aa protein might be a feasible male contraceptive which could affect sperm production without disturbing hormone levels.

关键词： follicle-stimulating hormone receptor prokaryotic recombinant expression male contraception vaccine

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