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Abstract

Volume 16, Issue 4 (July 2014) 16, 623–630; 10.4103/1008-682X.125910

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

Cheng Xu1, Ying-Chun Li2, Hua Yang3, Yan Long4, Min-Jian Chen1, Yu-Feng Qin1, Yan-Kai Xia1, Ling Song1, Ai-Hua Gu1, Xin-Ru Wang1

1 State Key Laboratory of Reproductive Medicine, Institute of Toxicology; Key Laboratory of Modern Toxicology of Ministry of Education, School of Public Health, Nanjing, China, China
2 Department of Physiology, Nanjing Medical University, Nanjing, China
3 Yangzhou Center for Disease Control and Prevention, Yangzhou, China
4 Department of Pharmacology, China Pharmaceutical University, Nanjing, China

Correspondence: Dr. AH Gu (aihuagu@njmu.edu.cn) or Dr. XR Wang (xrwang@njmu.edu.cn)

Received: 05 September 2013; Revised: 10 November 2013;Accepted: 27 December 2013

Abstract

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 coli strain BL21 Star TM (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.

Keywords: follicle-stimulating hormone receptor; prokaryotic recombinant expression; male contraception; vaccine

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Asian Journal of Andrology CN 31-1795/R ISSN 1008-682X  Copyright © 2023  Shanghai Materia Medica, Chinese Academy of Sciences.  All rights reserved.