Immunological response of female macaques to the PH-20 sperm protein following injection of recombinant proteins or synthesized peptides

X. Deng, Stuart A Meyers, Theodore L Tollner, A. I. Yudin, P. D. Primakoff, D. N. He, J. W. Overstreet

Research output: Contribution to journalArticle

14 Citations (Scopus)

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.

Original languageEnglish (US)
Pages (from-to)93-115
Number of pages23
JournalJournal of Reproductive Immunology
Volume54
Issue number1-2
DOIs
StatePublished - 2002

Fingerprint

Macaca
Recombinant Proteins
Peptides
Injections
Spermatozoa
Amino Acids
Contraceptive Vaccines
hyaluronidase PH-20
Antigens
Tetanus Toxoid
Antibodies
Tetanus
Indirect Fluorescent Antibody Technique
Fertilization
Gold
Abscess
Lysine
Antibody Formation
Immunization
Vaccines

Keywords

  • Contraception
  • Immunization
  • Macaque
  • PH-20
  • Sperm

ASJC Scopus subject areas

  • Immunology
  • Reproductive Medicine

Cite this

Immunological response of female macaques to the PH-20 sperm protein following injection of recombinant proteins or synthesized peptides. / Deng, X.; Meyers, Stuart A; Tollner, Theodore L; Yudin, A. I.; Primakoff, P. D.; He, D. N.; Overstreet, J. W.

In: Journal of Reproductive Immunology, Vol. 54, No. 1-2, 2002, p. 93-115.

Research output: Contribution to journalArticle

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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.",
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