The goal of this research program is to clone and characterize proteins which are located on the surface of the human sperm and to evaluate their immunocontraceptive potential. Attention will be directed to proteins which are auto- or iso-antigens and are recognized by sperm agglutinating antibodies. A computerized 2-D database based on migration of human sperm proteins in two dimensional gels has been developed. Within this database a subset of 98 human sperm surface proteins accessible to surface iodination and biotinylation has been identified. The locations of these surface proteins in two dimensional gels have been catalogued in a sperm protein encyclopedia. A subset of these sperm surface proteins will be microsequenced, and complete open reading frames will be obtained by molecular cloning. Unique sperm proteins will be characterized and evaluated as potential contraceptive immunogens according to strict criteria which include testis specificity and involvement in functions mediated at the sperm surface.
The specific aims are: 1) To microsequence human sperm surface proteins through a combination of Edman degradation and/or tandem mass spectrometry; 2) To clone and sequence the complete open reading frames for an estimated 20 cDNAs representing unique sperm surface proteins using a combination of molecular approaches; 3) To determine tissue specificity of the cloned cDNAs by Northern blot, RT-PCR, and immunohistochemical analyses on an organ bank of human and non- human primate mRNAs and tissues; 4) To confirm the sperm surface localization of candidate immunogens and test their contraceptive potential through in vitro functional assays; and 5) To conduct immunogenicity and fertility trials of several novel sperm surface proteins in mice and macaques. Achievement of these aims will increase our understanding of the molecular character of proteins exposed on the exterior of the human sperm surface and aid in selection of candidate antigens appropriate for human immunocontraception by immortalizing and manipulating the cDNAs encoding these proteins.

Project Start
2001-03-01
Project End
2003-02-28
Budget Start
Budget End
Support Year
10
Fiscal Year
2002
Total Cost
Indirect Cost
Name
University of Virginia
Department
Type
DUNS #
001910777
City
Charlottesville
State
VA
Country
United States
Zip Code
22904
Pires, Eusebio S; Hlavin, Callie; Macnamara, Ellen et al. (2013) SAS1B protein [ovastacin] shows temporal and spatial restriction to oocytes in several eutherian orders and initiates translation at the primary to secondary follicle transition. Dev Dyn 242:1405-26
Shumilin, Igor A; Cymborowski, Marcin; Chertihin, Olga et al. (2012) Identification of unknown protein function using metabolite cocktail screening. Structure 20:1715-25
Naaby-Hansen, Soren; Herr, John C (2010) Heat shock proteins on the human sperm surface. J Reprod Immunol 84:32-40
Naaby-Hansen, Soren; Diekman, Alan; Shetty, Jagathpala et al. (2010) Identification of calcium-binding proteins associated with the human sperm plasma membrane. Reprod Biol Endocrinol 8:6
Wolkowicz, M J; Digilio, L; Klotz, K et al. (2008) Equatorial segment protein (ESP) is a human alloantigen involved in sperm-egg binding and fusion. J Androl 29:272-82
Klotz, Kenneth L; Coppola, Michael A; Labrecque, Michel et al. (2008) Clinical and consumer trial performance of a sensitive immunodiagnostic home test that qualitatively detects low concentrations of sperm following vasectomy. J Urol 180:2569-76
Kurth, Barbara E; Digilio, Laura; Snow, Phillip et al. (2008) Immunogenicity of a multi-component recombinant human acrosomal protein vaccine in female Macaca fascicularis. J Reprod Immunol 77:126-41
Herr, John C; Chertihin, Olga; Digilio, Laura et al. (2008) Distribution of RNA binding protein MOEP19 in the oocyte cortex and early embryo indicates pre-patterning related to blastomere polarity and trophectoderm specification. Dev Biol 314:300-16
Shetty, Jagathpala; Bronson, Richard A; Herr, John C (2008) Human sperm protein encyclopedia and alloantigen index: mining novel allo-antigens using sera from ASA-positive infertile patients and vasectomized men. J Reprod Immunol 77:23-31
Xu, Bingfang; Hao, Zhonglin; Jha, Kula N et al. (2008) TSKS concentrates in spermatid centrioles during flagellogenesis. Dev Biol 319:201-10

Showing the most recent 10 out of 121 publications