The preparation and application of N-terminal 57 amino acid protein of the follicle-stimulating hormone receptor as a candidate male contraceptive vaccine.
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.
Cheng
C
Xu
Ying-Chun
YC
Li
Hua
H
Yang
Yan
Y
Long
Min-Jian
MJ
Chen
Yu-Feng
YF
Qin
Yan-Kai
YK
Xia
Ling
L
Song
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.
Ai-Hua
AH
Gu
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.
Xin-Ru
XR
Wang
D000818:Animals; D003272:Contraceptive Agents, Male / Q000008:administration & dosage*; D005260:Female; D006801:Humans; D008255:Macaca radiata; D008297:Male; D011247:Pregnancy; D018873:Pregnancy Rate; D011962:Receptors, FSH / Q000378:metabolism / Q000627:therapeutic use*; D012708:Sertoli Cells / Q000187:drug effects; D013091:Spermatogenesis / Q000187:drug effects*; D013094:Spermatozoa / Q000187:drug effects; D022642:Vaccines, Contraceptive / Q000008:administration & dosage*
D003272:Contraceptive Agents, Male; D011962:Receptors, FSH; D022642:Vaccines, Contraceptive