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Follicle-stimulating hormone (FSH) via interaction with G-protein coupled specific receptors plays a central role in the control of gametogenesis in mammals of both sexes. In females, FSH is crucial for follicle growth, follicle maturation and ovulation. FSH receptors, together with luteinizing hormone-chorionic gonadotropin and thyrotropin recepto...
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Glycoprotein receptors are a subfamily of G-protein coupled receptors, including the follicle hormone (FSH) receptor (FSHR), thyroid-stimulating hormone receptor (TSH), and luteinizing/chorionic gonadotrophin hormone receptor (LHCGR). These receptors display common structural features such as a prominent extracellular domain, with a leucine-rich re...
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... Although antibodies against fragments of hCG have been tested as a contraceptive vaccine [116], this approach was terminated due to a lack of efficacy. Immunization against fragments of FSH-ß [117] or FSH receptor [118] was also proposed as nonsteroidal contraceptives in both males and females. However, these approaches also suffered from lack of efficacy and uncertain long-term outcomes. ...
Gonadotropins belong to the family of dimeric glycoprotein hormones and regulate gonadal physiology mediated by G protein-coupled, seven-transmembrane receptors. These glycoprotein hormones are widely used in the clinic to promote ovarian follicle development and for treating some cases of male infertility. We traced the co-evolution of dimeric gonadotropin hormones and their receptors, together with thyrotropin and its receptor. We updated recent findings on human genetic variants of these genes and their association with dizygotic twining, polycystic ovarian syndrome, primary ovarian insufficiency, male-limited precocious puberty and infertility. In addition to the known physiological roles of gonadotropin-receptor signaling in gonadal tissues, we also discussed emerging understanding of extra-gonadal functions of gonadotropins in bones and adipose tissues, together with recent advances in in vivo imaging of gonadotropin receptors in live animals. Recent development of gonadotropin receptor agonists and antagonists were summarized with an emphasis on the development of functional antagonists for FSH receptors to alleviate osteoporosis and obesity associated with menopause.
... The applicative potential of anti-FSHR antibodies was demonstrated in different models. The cAMP production in cell lines overexpressing the FSH receptor was successfully antagonized by antibodies raised against its N-term [2] and its activity in vivo was neutralized by antibodies directed against the thumb region close to the active sulfotyrosine [4]. ...
... Since these antibodies have usually antagonist behavior, it is probable that their actual epitope overlaps the sequence 285-309 that corresponds to the region bound by the hormone FSH [4]. Only the immunization with decapeptides corresponding to the N-terminal FSHR ectodomain allowed for the production of agonist antibodies with clear stimulatory effect [2]. Otherwise, despite the increasing interest for FSHR in oncological diagnostic for both immunohistochemistry and imaging applications [9,14e17,27], no other antibody has been characterized for its binding features and has been proposed for in vivo treatment or to deliver either drugs or imaging probes. ...
Antibodies are essential reagents that are increasingly used in diagnostics and therapy. Their specificity and capacity to recognize their native antigen are critical characteristics for their in vivo application. Follicle-stimulating hormone receptor is a GPCR protein regulating ovarian follicular maturation and spermatogenesis. Recently, its potentiality as a cancer biomarker has been demonstrated but no antibody suitable for in vivo tumor targeting and treatment has been characterized so far. In this paper we describe the first successful attempt to recover recombinant antibodies against the FSHR and that: i) are directly panned from a pre-immune library using whole cells expressing the target receptor at their surface; ii) show inhibitory activity towards the FSH-induced cAMP accumulation; iii) do not share the same epitope with the natural binder FSH; iv) can be produced inexpensively as mono- or bivalent functional molecules in the bacterial cytoplasm. We expect that the proposed biopanning strategy will be profitable to identify useful functional antibodies for further members of the GPCR class.
... Jean-Jacques Remy's group produced recombinant filamentous phages displaying three overlapping Nterminal decapeptides of FSHR. When administered to animals they induced neutralizing antibodies against FSHR, blocking FSH-FSHR interaction [76,77]. The constructs were tested in several mammalian species and in animal groups of different sexual maturity. ...
Background:
Population control of domestic, wild, invasive, and captive animal species is a global issue of importance to public health, animal welfare and the economy. There is pressing need for effective, safe, and inexpensive contraceptive technologies to address this problem. Contraceptive vaccines, designed to stimulate the immune system in order to block critical reproductive events and suppress fertility, may provide a solution. Filamentous bacteriophages can be used as platforms for development of such vaccines.
Objective:
In this review authors highlight structural and immunogenic properties of filamentous phages, and discuss applications of phage-peptide vaccines for advancement of immunocontraception technology in animals.
Results:
Phages can be engineered to display fusion (non-phage) peptides as coat proteins. Such modifications can be accomplished via genetic manipulation of phage DNA, or by chemical conjugation of synthetic peptides to phage surface proteins. Phage fusions with antigenic determinants induce humoral as well as cell-mediated immune responses in animals, making them attractive as vaccines. Additional advantages of the phage platform include environmental stability, low cost, and safety for immunized animals and those administering the vaccines.
Conclusion:
Filamentous phages are viable platforms for vaccine development that can be engineered with molecular and organismal specificity. Phage-based vaccines can be produced in abundance at low cost, are environmentally stable, and are immunogenic when administered via multiple routes. These features are essential for a contraceptive vaccine to be operationally practical in animal applications. Adaptability of the phage platform also makes it attractive for design of human immunocontraceptive agents.
... Both of the approaches, cloning in a phage vector or selecting from a random phage display library, were used in the past successfully for generation of phages that modify animal reproductive functions. To develop a contraceptive, Abdennebi et al. (1999) produced recombinant filamentous phages displaying three overlapping N-terminal decapeptides of the follicle-stimulating hormone receptor (FSHR). The constructs were tested in several mammalian species and in animal groups of different sexual maturity. ...
... When injected in animals, they induced anti-FSHR immunity capable of inhibiting follicle-stimulating hormone binding. The vaccinations were shown to impair fertility in adult mice and inhibit ovulation rates in ewes and to induce infertility in adult male bonnet monkeys (Abdennebi et al., 1999;Rao et al., 2004). The same constructs were employed for immunization of male mice and goats at the prepubertal stage (Abdennebi et al., 2003). ...
Phage display is based on genetic engineering of phage coat proteins resulting in fusion peptides displayed on the surface of phage particles. The technology is widely used for generation of phages with novel characteristics for numerous applications in biomedicine and far beyond. The focus of this study was on development of phage-peptide constructs that stimulate production of antibodies against gonadotropin releasing hormone (GnRH). Phage-peptide constructs that elicit production of neutralizing GnRH antibodies can be used for anti-fertility and anti-cancer applications. Phage-GnRH constructs were generated via selection from a phage display library using several types of GnRH antibodies as selection targets. Such phage constructs were characterized for sequence similarities to GnRH peptide and frequency of their occurrence in the selection rounds. Five of the constructs with suitable characteristics were tested in mice as a single dose 5×10(11) virions (vir) vaccine and were found to be able to stimulate production of GnRH-specific antibodies, but not to suppress testosterone (indirect indicator of GnRH antibody neutralizing properties). Next, one of the constructs was tested at a higher dose of 2×10(12) vir per mouse in combination with a poly(lactide- co -glycolide) (PLGA)-based adjuvant. This resulted in multifold increase in GnRH antibody production and significant reduction of serum testosterone, indicating that antibodies produced in response to the phage-GnRH immunization possess neutralizing properties. To achieve optimal immune responses for desired applications, phage-GnRH constructs can be modified with respect to flanking sequences of GnRH-like peptides displayed on phage. Anticipated therapeutic effects also might be attained using optimized phage doses, a combination of several constructs in a single treatment, or application of adjuvants and advanced phage delivery systems.
... Several strategies for phage-based contraceptive vaccines have been proposed for control of animal populations. For example, immunization with phage displaying follicle-stimulating hormone peptides on pVIII elicited antibodies that impaired the fertility of mice and ewes (Abdennebi et al., 1999). Phage displaying or chemically TABLE 2 | Studies using filamentous phage as an immunogenic carrier for peptide B-cell epitopes. ...
For the past 25 years, phage display technology has been an invaluable tool for studies of protein-protein interactions. However, the inherent biological, biochemical, and biophysical properties of filamentous bacteriophage, as well as the ease of its genetic manipulation, also make it an attractive platform outside the traditional phage display canon. This review will focus on the unique properties of the filamentous bacteriophage and highlight its diverse applications in current research. Particular emphases are placed on: (i) the advantages of the phage as a vaccine carrier, including its high immunogenicity, relative antigenic simplicity and ability to activate a range of immune responses, (ii) the phage's potential as a prophylactic and therapeutic agent for infectious and chronic diseases, (iii) the regularity of the virion major coat protein lattice, which enables a variety of bioconjugation and surface chemistry applications, particularly in nanomaterials, and (iv) the phage's large population sizes and fast generation times, which make it an excellent model system for directed protein evolution. Despite their ubiquity in the biosphere, metagenomics work is just beginning to explore the ecology of filamentous and non-filamentous phage, and their role in the evolution of bacterial populations. Thus, the filamentous phage represents a robust, inexpensive, and versatile microorganism whose bioengineering applications continue to expand in new directions, although its limitations in some spheres impose obstacles to its widespread adoption and use.
... Immunization of prepubertal BALB/c male mice with phages displaying the FSHR specific decapeptide (amino acids 18-27) was shown to be an effective and reversible male contraceptive (Remy et al., 1996). Abdennebi et al. (1999) engineered filamentous phages displaying the peptides 18-27 (A), 25-34 (B) and 29-38 (C). They observed that Anti-C IgG had FSH-like agonistic activity and the peptidic vaccines A and B showed reversible inhibition of ovulation in ewes and impaired fertility in female mice. ...
... 6,7 The ECD of FSH receptor (FSHR) has also been considered a suitable target for a male vaccine, as it is expressed exclusively on testicular Sertoli cells. 8,9 With respect to gamete-associated components, research has focused on the relationship between sperm antigens and fertility in past decades. Among numerous molecular candidates related to sperm, epididymis protease inhibitor (Eppin) is a promising candidate. ...
In our previous study on adult male mice, we had identified one immunodominant epitope in hEppin and three epitopes in hFSHR that caused fertility inhibition. But it only demonstrated a moderate inhibitory effect on fertility, and the antifertility effect was unsatisfactory.
Based on the protein prime-peptide boost inoculation modalities, we further investigated whether the antifertility capacity could be enhanced by a combined immunization with the two antigens.
The results displayed a enhanced suppressed fertility (F2EP2C 6.67%) in male mice similar to that seen after four separate administrations of the two proteins (F12E-4 5%). The most likely mechanism by which this antifertility efficacy was achieved was probably through the production of antibodies that led not only to impairment of spermatogenesis but also to inhibition of sperm motility. Moreover, this treatment also induced high concentrations of neutralizing antibodies which were secreted into the lumen of the epididymis.
Thus, a combination immunization with hFSHR and hEppin enhanced the contraceptive effects and may provide a better means of immunocontraception.
© 2015 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.
... Studies on development of phagebased vaccines explored preparations for treatment of melanoma (Eriksson et al., 2007), HIV (De Berardinis et al., 2003, Alzheimer's disease (Frenkel et al., 2003), candidiasis (Wang et al., 2006;Yang et al., 2007), and rabies (Houimel and Dellagi, 2009). Furthermore, recombinant phages displaying decapeptides of follicle-stimulating hormone receptor were shown to impair fertility in mice and inhibit ovulation rates in ewes (Abdennebi et al., 1999) and to induce infertility in adult male bonnet monkeys (Abdennebi et al., 1999;Rao et al., 2004), suggesting the potential use of phage-based vaccines for immunocontraception. Several studies indicated that vaccines based on phage have the potentials to be effective immunogens after oral administration (Delmastro et al., 1997;Jensen-jarolim et al., 1998;Zuercher et al., 2000). ...
... Studies on development of phagebased vaccines explored preparations for treatment of melanoma (Eriksson et al., 2007), HIV (De Berardinis et al., 2003, Alzheimer's disease (Frenkel et al., 2003), candidiasis (Wang et al., 2006;Yang et al., 2007), and rabies (Houimel and Dellagi, 2009). Furthermore, recombinant phages displaying decapeptides of follicle-stimulating hormone receptor were shown to impair fertility in mice and inhibit ovulation rates in ewes (Abdennebi et al., 1999) and to induce infertility in adult male bonnet monkeys (Abdennebi et al., 1999;Rao et al., 2004), suggesting the potential use of phage-based vaccines for immunocontraception. Several studies indicated that vaccines based on phage have the potentials to be effective immunogens after oral administration (Delmastro et al., 1997;Jensen-jarolim et al., 1998;Zuercher et al., 2000). ...
Multiple phage-peptide constructs, where the peptides mimic sperm epitopes that bind to zona pellucida (ZP) proteins, were generated via selection from a phage display library using a novel approach. Selections were designed to allow for identification of ZP-binding phage clones with potential species-specific properties, an important feature for wildlife oral vaccines as the goal is to control overpopulation of a target species while not affecting non-target species' reproduction. Six phage-peptide antigens were injected intramuscularly into pigs and corresponding immune responses evaluated. Administration of the antigens into pigs stimulated production of anti-peptide antibodies, which were shown to act as anti-sperm antibodies. Potentially, such anti-sperm antibodies could interfere with sperm delivery or function in the male or female genital tract, leading to contraceptive effects. Staining of semen samples collected from different mammalian species, including pig, cat, dog, bull, and mouse, with anti-sera from pigs immunized with ZP-binding phage allowed identification of phage-peptide constructs with different levels of species specificity. Based on the intensity of the immune responses and specificity of these responses in different species, two of the antigens with fusion peptide sequences GEGGYGSHD and GQQGLNGDS were recognized as the most promising candidates for development of contraceptive vaccines for wild pigs.
... In mammals, we have demonstrated in previous studies that targeting specific regions of LH and FSH receptors through the immune pathway induces impairment of adult fertility (Remy et al., 1996;Abdennebi et al., 1999;Rao et al., 2004) and delays sexual maturity (Abdennebi et al., 2003). In the present study, we used the same phage display strategy to express multiple copies of rainbow trout FSH and LH receptor epitopes on the surface of phage particles. ...
... Due to these vaccine-specific inhibitory effects on different physiological processes, we can exclude an effect of the vector itself. As demonstrated in other species, injection of non-recombinant fd phages never induced any effect on reproductive parameters (Abdennebi et al., 1999). Furthermore, in a complementary experiment we verified that only the vaccination with phage displaying a gonadotropin receptor peptide had inhibitory effects on trout spermatogenesis, while phage displaying random peptides had no significant effect. ...
... Recombinant phages expressed 17 amino acids of the trout Lhr or Fshr: although these two sequences share four residues, it is far from being enough to induce cross-reactive antibodies. In previous studies in mammals, we demonstrated that immunization with such hybrid bacteriophages induced immunity against the targeted gonadotropin receptor (Abdennebi et al., 1999). To take into account the fact that the response type observed in a mammal is not necessarily conclusive for the situation in trout, we purified IgM from immunized fish and tested their ability to inhibit the response to FSH of COS-7 cells expressing the Fshr. ...
In fish, gonadotropin hormones FSH-GTH1 and LH-GTH2 are less specific for their cognate receptors than in mammals. The respective reproductive functions of fish LH and FSH are thus difficult to establish. We aimed to study the effect of specific antagonists of the two gonadotropin receptors on trout sexual maturation in both sexes by targeting specific regions of LH and FSH receptors, Lhr and Fshr. Filamentous phages displaying Lhr specific or Fshr specific decapeptides from the extracellular hormone binding domain were engineered. Recombinant phages were used as receptor-specific antagonistic vaccines. Male and female trouts were immunized with anti-LHR, anti-FSHR, anti-FSHR+LHR or adjuvant alone, through multiple injections over 8-24 weeks, starting at different stages of sexual maturation. The consequences of immunization on gonadal development were evaluated by determining gonad growth, by histological analysis of testis and ovaries at the end of the vaccination period and by measuring blood plasma sex steroids using radioimmunoassay. We show for the first time in fish that the anti-receptor vaccinations could have specific antagonistic effects on the development of the reproductive functions; while the anti-FSHR affected the sexual maturation of prepubertal males and delayed sperm production, the anti-LHR blocked vitellogenesis in females. In maturing males, the combined anti-FSHR+LHR vaccine inhibited spermatogenesis and affected steroidogenesis. In that case, the effects of the vaccine on spermatogenesis were transient and reversible when immunization was stopped. Such an immunological strategy to specifically and transiently inhibit a receptor provides a promising approach for discovering their specific functions; it could also lead to a new technology for controlling the onset of puberty in aquaculture species.
... In small ruminants, the placental transfer of EDCs during the prenatal period GD 60 -150 ensures the exposure of testes and hypothalamic-pituitary-testicular axis during critical stages of male fetal development (Brooks et al. 1996, Sweeney et al. 2000. Little is known about the regulation and crosstalk between LH and FSH and significant differences between species are demonstrated (Jansen et al. 1993, Gerhard et al. 1998, Abdennebi et al. 1999. Most evidence of the disruption of LH secretion by PCBs is derived from laboratory animal studies; for example, daily exposure of lactating rats to Aroclor 1242, reduced plasma testosterone and LH levels as well as a reduction in the number of Leydig cells in the testis. ...
In this study, female goats were orally exposed to PCB126 or PCB153, at 49 ng/kg body weight per day and 98 microg/kg body weight per day respectively, from gestational day 60 until delivery at approximately day 150. Exposure of the offspring continued via lactation until postnatal day 40. Reproductive toxicity in the male offspring was studied by the evaluation of conventional reproductive endpoints as well as flow cytometric analyses of spermatogenesis and sperm chromatin structure. PCB153-treated animals showed a significant smaller testis diameter in comparison to the control group. Neither of the treated groups showed differences for plasma FSH in comparison to controls. PCB153-treated animals differed significantly from the control group with respect to plasma LH and testosterone levels, whereas PCB126-treated animals only differed from the controls in plasma testosterone concentrations. Neither the PCB126 nor the PCB153 group differed from the controls with respect to the conventional sperm parameters or testis histology. A significant lower ratio of interstitium area to seminiferous tubules area and proportion of diploid testis cells were observed for the PCB153 group. Sperm from PCB153-treated animals showed a significantly higher percentage of sperm with damaged DNA. From the results of the present study it was concluded that PCB153 was able to induce alterations in reproductive endpoints related to the hypothalamic-pituitary-axis as well as to the testis. The effects observed in male kids after a long-term maternal exposure to PCB153 support the concept that exposure to endocrine-disrupting compounds during foetal development may lead to adverse reproductive effects in adult life.
