title: Immunological response of female macaques to the PH-20 sperm protein following injection of recombinant proteins or synthesized peptides.
abstract: Because of its location on the sperm surface and its multiple functions during fertilization, the PH-20 protein is a potential target for contraceptive vaccines. Cynomolgus macaques were immunized using four different adjuvants together with synthesized peptides or recombinant proteins representing selected regions of macaque PH-20. The synthesized peptide (amino acids 387-412, designated Peptide 4) was used as a linear molecule in a 1:1 ratio with a peptide sequence of tetanus toxoid, as well as a multiple antigenic peptide (MAP) matrix held together by scaffolding lysine residues. In the MAP construct, the ratio of Peptide 4 to tetanus peptide was 4:1. To circumvent the poor production of recombinant PH-20 in bacterial cells, two truncated forms of the molecule were expressed in Escherichia coli, G18 (encoding amino acids 143-510) and E10 (encoding amino acids 291-510). The adjuvants were Montanide ISA 51, Titermax Gold, Syntex adjuvant formulation (SAF), and QS-21. All of the antigen/adjuvant combinations produced significant immune responses as measured by ELISA. The circulating antibodies from immunized animals recognized macaque sperm surface PH-20 on Western blots and were shown by indirect immunofluorescence to bind to the surface of macaque sperm. Montanide and Titermax were associated with higher titers of anti-PH-20 antibodies than QS-21 and SAF adjuvants. Immunization with Titermax, however, resulted in sterile abscesses in 4 of 8 animals injected. We conclude that antigens derived from synthesized peptides and recombinant proteins representing selected regions of the PH-20 molecule can be used as vaccine components in combination with the adjuvant Montanide to elicit a significant sperm-directed antibody response in immunized macaques.
authors.affiliation: Division of Reproductive Biology, Department of Obstetrics and Gynecology, Institute of Toxicology and Environment Health, One Shields Avenue, University of California, Davis, CA 95616-8739, USA.
authors.forename: X
authors.initials: X
authors.lastname: Deng
authors.forename: S A
authors.initials: SA
authors.lastname: Meyers
authors.forename: T L
authors.initials: TL
authors.lastname: Tollner
authors.forename: A I
authors.initials: AI
authors.lastname: Yudin
authors.forename: P D
authors.initials: PD
authors.lastname: Primakoff
authors.forename: D N
authors.initials: DN
authors.lastname: He
authors.forename: J W
authors.initials: JW
authors.lastname: Overstreet
mesh_terms: D000119:Acetylmuramyl-Alanyl-Isoglutamine / Q000031:analogs & derivatives* / Q000494:pharmacology; D000276:Adjuvants, Immunologic / Q000494:pharmacology; D000595:Amino Acid Sequence; D000818:Animals; D000906:Antibodies / Q000097:blood; D015153:Blotting, Western; D015815:Cell Adhesion Molecules / Q000235:genetics / Q000276:immunology*; D003267:Contraception; D005260:Female; D006821:Hyaluronoglucosaminidase; D007114:Immunization; D008252:Macaca fascicularis; D008297:Male; D008969:Molecular Sequence Data; D011060:Poloxalene / Q000494:pharmacology; D011136:Polysorbates / Q000494:pharmacology; D011994:Recombinant Proteins / Q000276:immunology; D012503:Saponins / Q000494:pharmacology; D013185:Squalene / Q000031:analogs & derivatives* / Q000494:pharmacology
chemical_list: D000276:Adjuvants, Immunologic; D000906:Antibodies; D015815:Cell Adhesion Molecules; D011136:Polysorbates; D011994:Recombinant Proteins; D012503:Saponins; C077975:Syntex adjuvant formulation; C086752:TiterMax; D000119:Acetylmuramyl-Alanyl-Isoglutamine; C078785:saponin QA-21V1; D013185:Squalene; D011060:Poloxalene; D006821:Hyaluronoglucosaminidase; C066852:hyaluronidase PH-20