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Hum. Reprod. Advance Access published online on September 7, 2006
Human Reproduction, doi:10.1093/humrep/del068
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© The Author 2006. Published by Oxford University Press on behalf of the European Society of Human Reproduction and Embryology. All rights reserved. For Permissions, please email: journals.permissions@oxfordjournals.org
Received October 18, 2005
Revised January 6, 2006
Accepted February 10, 2006

Article

Immunogenicity study of recombinant human sperm-associated antigen 9 in bonnet macaque (Macaca radiata)

Ritu Rana 1, Nirmala Jagadish 1, Manoj Garg 1, Deepshikha Mishra 1, Neetu Dahiya 1, Dipak Chaurasiya 1, and Anil Suri 1 *

1 Genes and Proteins Laboratory, National Institute of Immunology, Aruna Asaf Ali Marg, New Delhi, India

* To whom correspondence should be addressed.
Anil Suri, E-mail: anil{at}nii.res.in


  Abstract

BACKGROUND: The human sperm-associated antigen 9 (hSPAG9) is of special interest attributing to the findings indicating that SPAG9 is an acrosomal molecule. SPAG9 is not only restricted to acrosomal compartment but also persists in equatorial segment post-acrosome reaction, which is a key location in sperm-egg interaction. METHODS AND RESULTS: Immunogenicity studies in macaques were carried out with recombinant hSPAG9 (rhSPAG9) adsorbed on alum, which resulted in high titres of anti-rhSPAG9 antibodies as determined by enzyme-linked immunosorbent assay (ELISA). Immunoblotting analysis employing anti-rhSPAG9 antibodies generated in monkeys indicated that antibodies specifically reacted with native SPAG9 from macaque and human sperm and rhSPAG9 protein. Furthermore, indirect immunofluorescence experiments demonstrated SPAG9 localization in the acrosomal compartment of macaque and human sperm. In addition, monkey antibodies against rhSPAG9 significantly inhibited the human spermatozoa adherence or penetration in zona-free hamster oocytes. CONCLUSION: These results demonstrate that rhSPAG9 adsorbed on alum is highly immunogenic in subhuman primate model and therefore represents a suitable sperm-based vaccine immunogen for fertility trials in macaque.

Keywords: bonnet macaque/human sperm/immunogenicity/sperm-associated antigen 9/sperm-egg interaction.
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