Kaul R, Afzalpurkar A, Gupta S K
Gamete Antigen Laboratory, National Institute of Immunology, New Delhi, India.
J Reprod Fertil Suppl. 1996;50:127-34.
Immunization of female bonnet monkeys (Macaca radiata) with pig zona pellucida glycoprotein ZP3 using adjuvants permissible for human use leads to infertility. Fifty per cent of the animals conceived after antibody titres declined. Animals that failed to regain their fertility did not show any obvious ovarian changes. As part of the attempt to design an effective immunocontraceptive vaccine based on synthetic peptides, the epitopes recognized by monoclonal antibodies (mAb) against pig ZP3 alpha and ZP3 beta and possessing contraceptive efficacy in vitro were mapped. These studies identified an N-blocked decapeptide from the N-terminus corresponding to 23-32 amino acids of the precursor protein of pig ZP3 beta. As an alternative approach to using synthetic peptides, DNA encoding bonnet monkey ZP3 was cloned and sequenced. The deduced primary amino acid sequence revealed an overall similarity of 93.9% with human ZP3. Bonnet monkey ZP3 corresponding to an internal 975 nucleotide fragment excluding the N-terminus signal sequence and the C-terminus transmembrane domain has been expressed in Escherichia coli. These results will assist in the design of an immunocontraceptive vaccine based on either synthetic peptides or recombinant glycoproteins or proteins corresponding to ZP.
使用人类可用佐剂用猪透明带糖蛋白ZP3对雌性帽猴(食蟹猴)进行免疫会导致不育。抗体滴度下降后,50%的动物受孕。未能恢复生育能力的动物未表现出任何明显的卵巢变化。作为基于合成肽设计有效免疫避孕疫苗尝试的一部分,绘制了针对猪ZP3α和ZP3β且在体外具有避孕效果的单克隆抗体(mAb)识别的表位。这些研究从猪ZP3β前体蛋白对应于23 - 32个氨基酸的N端鉴定出一种N端封闭的十肽。作为使用合成肽的替代方法,克隆并测序了编码帽猴ZP3的DNA。推导的一级氨基酸序列显示与人类ZP3的总体相似性为93.9%。对应于一个内部975核苷酸片段(不包括N端信号序列和C端跨膜结构域)的帽猴ZP3已在大肠杆菌中表达。这些结果将有助于基于合成肽、重组糖蛋白或与ZP对应的蛋白质设计免疫避孕疫苗。