Novel Sperm and Gonadotropin-Releasing Hormone-Based Recombinant Fusion Protein: Achievement of 100% Contraceptive Efficacy by Co-Immunization of Male and Female Mice

摘要

Improvements in long-term female contraception can be achieved by vaccinating with sperm-derived proteins. Here, recombinant proteins comprising either (i) N- (amino acid residues 1-80) or C- (amino acid residues 76-126) terminal fragments of mouse sperm protein 17 (Sp17) fused to the promiscuous T non-B cell epitope of tetanus toxoid (TT), amino acid residues 830-844 followed by di-lysine linker (KK) (TT-KK-Sp17(N) or TT-KK-Sp17(C), respectively) or (ii) mouse equatorin (amino acid residues 21-185) fused to the T non-B cell epitope of bovine RNase (amino acid residues 94-104) were expressed in Escherichia coli. Immunization of female FVB/J mice, using alum as an adjuvant, led to the generation of high antibody titers against the above proteins. Antibodies against both N- and C-terminal fragments of Sp17 reacted with the entire capacitated mouse spermatozoa, whereas those against equatorin reacted exclusively with the equatorial region. Despite the reactivity of all immune sera, only sera from mice immunized with TT-KK-Sp17(N) and TT-KK-Sp17(C) significantly reduced mouse in vitro fertilization. Mating studies of the immunized females with un-immunized male mice revealed the highest infertility in the TT-KK-Sp17(C)-immunized group. In an attempt to further boost the immune response, the C-terminal fragment of Sp17 was expressed as fusion protein with a tandemrepeat of gonadotropin-releasing hormone (GnRH) (Sp17(C)-GnRH(2)). Immunization of both male and female mice with Sp17(C)-GnRH(2) led to higher contraceptive efficacy compared to mice immunized with TT-KK-Sp17(C). Interestingly, mating studies wherein partners were both immunized with Sp17(C)-GnRH(2) showed a complete failure of female mice to conceive. Thus, immunization of both males and females with Sp17(C)-GnRH(2) has the potential to increase contraceptive efficacy. (C) 2016 Wiley Periodicals, Inc.