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ISSN 2052-1472 (online)

Reproduction Abstracts (2014) 1 P151 | DOI: 10.1530/repabs.1.P151

Detection of ZP-autoantibody in the sera from infertile women using a human multiepitope peptide as antigen

Wan Xiang Xu1, Jian Wang1, Weijie Zhu2, Xiaoxue Chang3, Xiaoxi Sun4, Satish K Gupta5, Yaping He1, Haiping Tang1, Huijuan Shi1, Shaohua Gu6, Chaoneng Ji7 & Yi Xie6

1Shanghai Institute of Planned Parenthood Research, Shanghai, China; 2Institute of Reproductive Immunology, Jinan University, Guangzhou, China; 3Medical School, Henan University of Science and Technology, Luoyang, China; 4Shanghai Medical College, Obstetrics and Gynecology Hospital, Fudan University, Shanghai, China; 5National Institute of Immunology, New Delhi, India; 6School of Life Science, Institute of Genetics, Fudan University, Shanghai, China; 7State Key Laboratory of Genetic Engineering, School of Life Sciences, Institute of Genetics, Fudan University, Shanghai 200433, People’s Republic of China.

Background: The role of autoantibodies against zona pellucida (ZP) in infertility and success of IVF is debatable. In the present study, a novel recombinant human zona protein is used to address this issue.

Materials and methods: Fourteen serum samples positive for porcine ZP, six from infertile patients, and 20 from healthy women were analysed. Sera were tested for their reactivity against a recombinant multi-epitope peptide (named ZPCP7) composed of ten predictable antigenic peptides from human ZP proteins by ELISA. In addition, seven identified fine epitope peptides (fused with a GST188 carrier protein) of human ZP3 and ZP4 were used as antigens for western blotting.

Results and discussion: Eight of 14 porcine ZP antibody positive sera and one out of six testing infertile patients showed positive reaction in ELISA when all sera were tested at 1:2560 dilution. Further, out of nine serum samples seven also showed reactivity to three to five of epitope peptides fused with truncated GST188 carrier protein in western blot respectively. Thus, seven sera samples were finally determined as ZP antibody positive in our two steps method of analysis. Also, the positive serum sample (no. S3) was able to recognize native human ZP in indirect immunofluorescence experiment. These results provide evidence that ZP antibodies are associated with infertility. The use of recombinant multi-epitope peptide based on human zona proteins may also overcome the problem of false positivity observed using porcine ZP as an antigen.