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Follicle-stimulating hormone (FSH) has been newly demonstrated to play a great role in promoting fat accumulation, providing a potential to target FSH for controlling fat accumulation and treating obesity. A short, 13-amino acid of FSHβ (FSHβ13AA) was indentified to be the FSH receptor-binding epitope in both humans and mice. By conservation analysis, we found such FSHβ13AA is highly conserved across species. Accordingly, we designed a new FSH antigen by synthesizing a tandem of FSHβ13AA (LVYKDPARPNIQK) and then conjugating it to ovalbumin (FSHβ13AA-T-OVA). Then, we tested its efficacy in suppressing fat accumulation in both ovariectomized and intact mouse models. Vaccination with this novel antigen emulsified in mild adjuvant, Specol, was highly effective in preventing ovariectomy-induced body weight gain and fat accumulation in mice (P < 0.01). Mechanistically, FSH vaccination treatment inhibited lipid biosynthesis by inactivating PPARγ adipogenic signaling pathway and simultaneously enhanced adipocyte themogenesis via upregulating UCP1 expression in both visceral and subcutaneous adipose tissues. Moreover, injection of this novel FSH vaccine also substantially reduced (P < 0.05) fat accumulation in both intact male and female mice. These actions result from the specific binding of the generated antibody to the β-subunit to block its action, rather than lowering the circulating levels of FSH, as evidenced by nearly no alterations in serum FSH levels in mice following FSH vaccination. Overall, we developed a novel FSH antigen and vaccine, and demonstrated it is highly efficacious in suppressing fat accumulation., (Copyright © 2020 Elsevier Inc. All rights reserved.)

Research Question: To evaluate the immunogenicity of follitropin delta in repeated ovarian stimulation., Design: Controlled, assessor-blind trial in IVF/intracytoplasmic sperm injection patients undergoing repeated cycles of ovarian stimulation (cycles 2 and 3), following initial stimulation with follitropin delta or follitropin alfa (cycle 1) in a preceding randomized trial. In cycles 2 and 3, 513 and 188 women, respectively, were treated as randomized in cycle 1, with dosing based on ovarian response in the previous cycle., Results: The incidence of treatment-induced anti-FSH antibodies with follitropin delta was 0.8% and 1.1% in cycles 2 and 3, respectively, which was similar to the incidence in cycle 1 (1.1%). No antibodies were of neutralizing capacity. Women with pre-existing anti-FSH antibodies were safely treated with follitropin delta without boosting an immune response. Treatment with follitropin delta and follitropin alfa gave similar outcomes for mean number of oocytes retrieved (9.2 versus 8.6 [cycle 2]; 8.3 versus 8.9 [cycle 3]), ongoing pregnancy (27.8% versus 25.7%; 27.4% versus 28.0%) and live birth rates (27.4% versus 25.3%; 26.3% versus 26.9%). The presence of anti-FSH antibodies did not affect the ovarian response., Conclusions: The trial demonstrated the low immunogenicity potential of follitropin delta in repeated ovarian stimulation, and confirmed the appropriateness of the follitropin delta dosing regimen in repeated cycles, with documented efficacy and safety., (Copyright © 2018 The Authors. Published by Elsevier Ltd.. All rights reserved.)

In mammals, recovery of oocytes by laparoscopic ovum pick-up (LOPU) coupled with in vitro production (IVP) of embryos represents a promising strategy for both amplification and genetic management of sparse animals from captive endangered wild species. As integrated technique developed mainly for domestic livestock, LOPU-IVP requires several studies to set up protocols for follicular stimulation or optimization of IVP before envisaging successful transposition to wild species. In deer, many endangered subspecies would be potentially concerned by applying such an approach using common subspecies for protocols optimization. The aim of the present study was to assess efficiency of follicle stimulation using ovine FSH (oFSH) for recovery of oocytes by LOPU in common sika deer (Cervus nippon nippon) before transposition of an optimized methodology for IVP of embryos from endangered Vietnamese sika deer hinds (Cervus nippon pseudaxis). In common sika deer, two doses of oFSH (0.25 and 0.5 U) and two frequencies of administration (12 and 24 h) were compared by monitoring of subsequent ovarian response, quality of oocytes recovered by LOPU, and in vitro developmental competence. In a first experiment, the dose of oFSH administered did not significantly affect the total number of follicles aspirated per hind per session (8.6 ± 1.0 vs. 8.2 ± 1.6 with 0.5 vs. 0.25 U oFSH, respectively; not significant). In a second experiment, frequency of 0.25 U oFSH administration did not affect ovarian response. Efficiency of IVP determined on blastocysts rates after in vitro maturation, fertilization, and development in oviduct epithelial cells coculture was increased when FSH was administered at 12-h intervals. Immune response after several follicular stimulations was detected against exogenous oFSH in plasma from the majority of sika deer hinds but was not associated with decreased ovarian response. When 0.25 U oFSH was administered at 12-h intervals to Vietnamese sika deer (N = 4), good quality cumulus oocyte complexes with complete and compact cumulus investments were recovered allowing a high cleavage rate after in vitro maturation and fertilization. Development to the blastocyst stage occurred in a high proportion (30% of oocytes) after coculture with ovine epithelial cells allowing cryobanking of transferable embryos from Vietnamese sika deer. These results confirm that LOPU-IVF after ovarian stimulation with oFSH may be a successful tool for cryobanking transferable embryos from endangered sika deer subspecies., (Copyright © 2012 Elsevier Inc. All rights reserved.)

Aims: Anti-follicle-stimulating hormone (FSH) autoantibody was found to highly correlate with oligospermia and asthenospermia, but the actual effect of FSH autoantibody on spermatogenesis is still unknown., Main Methods: In this study, 21-day rats were immunized seven times with FSH peptides linked with Keyhole Limpet Hemocyanin (KLH) (experimental group) or KLH (control group) every 2 weeks. Luteinizing hormone (LH) and inhibin B level in the immunized rat sera were measured by enzyme-linked immunosorbent assay (ELISA). Apoptosis of spermatogenic cells in the testis was detected by in situ end labeling method (TUNEL), and the mRNAs of Bax, Bcl-2 and Caspase-3 in testis were detected by fluorescent Quantitative PCR., Key Findings: Compared with the control, serum inhibin B level was significantly decreased at all time points (34.49%, 23.20%, and 37.00%) (p<0.05). There was no difference in the serum LH level between experimental and control groups. FSH peptide immunization increased the apoptosis of spermatogenic cells in the testis that was associated with an imbalance of Bax and Bcl-2 expression and upregulation of Caspase-3., Significance: These results suggest that FSH autoantibody could cause the reduction of inhibin B, thereby inducing hypospermatogenesis via augment of spermatogenic cell apoptosis., (Copyright © 2012 Elsevier Inc. All rights reserved.)

Single- and multi-analyte detection of two gonadotropic hormones (follicle stimulating hormone (hFSH) and luteinizing hormone (hLH)) was achieved by a Surface Plasmon Resonance (SPR) immunoassay on untreated human urine samples. Multi-analyte detection was accomplished using two alternative formats which are based in the individual or simultaneous immobilization of the hormones on the sensor surface. The lowest detection limit for both hormones in urine was found to be 1 ng mL(-1), which in international units (IU) in terms of the World Health Organization (WHO) standards represents 8 mIU mL(-1) of hLH and 14 mIU mL(-1) of hFSH, respectively. The reliability of the assay was demonstrated by intra- and inter-assay variabilities < 6%, chip-to-chip variabilities < 5%, recoveries in the range of 80-120% and stability of the sensor response through more than 100 measurements. The sensitivity of this biosensing methodology renders it in a useful technique for the diagnosis of reproductive disorders, as well as for fertility monitoring.

The growth and function of the epididymis are regulated by testosterone produced by Leydig cells in the testes. In the present study it was observed that neutralization of endogenous follicle stimulating hormone (FSH) in immature rats using a highly specific antiserum to ovine FSH resulted in changes in the histology of the epididymis along with a decrease (50-60%) in its weight compared with the normal serum-treated controls. These changes were observed in both rat and monkey epididymis without any decrease in serum testosterone, on which epididymis is known to be dependent. A detailed study was therefore carried out on the effects of deprivation of FSH or testosterone on the histology of epididymis. The changes in epididymal histology following FSH deprivation included a decrease in the size of the tubule lumen in the rat as well as in the adult male bonnet monkey in which the antiserum against ovine FSH was raised. Intensive vacuolization and uneven surface of the luminal epithelium was also observed. In contrast, the effect of deprivation of testosterone support by way of administration of LH antiserum or fiutamide resulted in a decrease in the size of the lumen and degenerative changes. These results suggest that cauda epididymidis is a target for FSH action.

FSH is required for spontaneous folliculogenesis and is widely used in ovarian stimulation in IVF. Previously, increased concentrations of antibodies against FSH (anti-FSH) have been demonstrated in infertile women. This study aimed to: (i) assess the possible association of anti-FSH with an adverse outcome of IVF with regard to clinical parameters characterizing the ovarian reserve; and (ii) compare serum and follicular fluid (FF) anti-FSH concentrations in relation to follicle size and endocrine markers. IVF patients (n = 182) subjected to gonadotrophin-releasing hormone-antagonist protocol were assessed for anti-FSH using enzyme-linked immunosorbent assay. Increased concentrations of serum anti-FSH immunoglobulin (Ig)G and IgA were associated with impaired ovarian stimulation outcome, with cut-off values <1.0 arbitrary units predicting poor ovarian response (

Spermatogenesis is a complex process, and previous studies have clearly established the role for testosterone in its maintenance. However, the role of FSH remains controversial, although several lines of evidence suggest its importance in initiation of spermatogenesis. In the present study, the relative roles of FSH and LH have been evaluated using specific antisera capable of neutralizing endogenous hormones in adult male rats following ethanedimethylsulphonate (EDS) treatment. Restoration of spermatogenesis in EDS-treated rats was monitored following FSH or LH deprivation by histological analysis and flow cytometry. Deprivation of FSH resulted in a reduction of seminiferous tubule diameter and spermatogonial number, which was much more drastic than that observed following LH deprivation. More importantly, FSH deprivation was associated with a significant reduction in the number of pachytene spermatocytes. These results provide evidence for a definite role for FSH in regulation of spermatogenesis, in addition to confirming the role of LH in spermatogenesis via testosterone.

The purpose of this study was to identify autoantigens contained in human ovary extracts. Serum samples from 36 infertile women with anti-ovary antibodies as detected with an ELISA technique were tested in Western blot against human ovary extracts. A reactive protein with a molecular mass matching that of the FSH was detected in 34 cases. These serum samples also reacted strongly in Western blot and ELISA with purified FSH and, in immunofluorescence, with pituitary cells. Using the Pepscan approach, with overlapping peptides matching the amino acid sequence of the human FSH beta-chain, several immunoreactive regions were evidenced. The 78-93 amino acid sequence of the human FSH beta-chain appeared as one of the major epitopes. Synthetic peptides of this region were prepared and demonstrated to react with human serum samples from women with anti-ovary antibodies. These data demonstrate that FSH can be an autoantigen, recognized by autoantibodies associated with infertility., ((c)2001 Elsevier Science.)

Overpopulation is a paramount global crisis, as it underlies virtually all major worldwide problems, including poverty, malnutrition, and warfare. It is imperative that proliferation be curbed by expediting reductions in fertility. This article discusses the demand for and acceptability of male contraceptives and the process of spermatogenesis. Endocrine control of spermatogenesis, and potential sites for disruption of male fertility, including steroid combinations and GnRH agonists and antagonists are reviewed also. Risks and benefits of these approaches are considered.

We investigated the importance of monoclonal antibody (MCA) purity and the input molar ratio of horseradish peroxidase (HRPO)/IgG used for MCA conjugation on various immunoenzymometric assay (IEMA) parameters. The sensitivity of IEMAs for human follicle stimulating hormone (hFSH), human chorionic gonadotropin (hCG) and the free alpha subunit of hCG (hCG alpha) could be increased up to 6-fold, whereas non-specific binding remained within tolerable limits (E less than 0.1), when MCAs purified by high performance liquid chromatography (HPLC) using a hydroxylapatite column (HPHT) were conjugated with an input molar HRPO/IgG ratio of four instead of the usual ratio of two.

Gonadotropic hormones which have lost peripheral sugar residues in their oligosaccharide chains display antagonistic properties and produce antibodies that fail to recognize the fully glycosylated hormone (agonist form). These polyclonal antibodies were separated by successive affinity chromatography on divinylsulfonyl-Sepharose coupled agonist and antagonist columns. The immunoglobulin G fraction from the agonist affinity columns recognizes both free agonist and antagonist in solution radioimmunoassays and also when these hormone forms are bound to receptors on gonadal cells. However, antagonist-specific antibodies recognize only the free antagonist in solution but not when it is receptor bound, implying that the conformation of the receptor-bound antagonist is different from that of the agonist. Affinity-purified antibodies against the different forms are useful in analyzing immunological and biological properties of the hormones. The studies with these glycosylated hormones serve as a useful model for other glycoproteins of pharmaceutical value.

Freezing and thawing of human serum produces alkaline changes in the pH. Weak buffers were of insufficient capacity to compensate for these pH changes. Thus the effect of serum pH on modified solid-phase and double-antibody radioimmunoassay (RIA) of LH and FSH was studied. Pooled postmenopausal serum standardized against the Second International Reference Preparation of Human Menopausal Urinary Gonadotropon was used as a laboratory standard. In the solid phase RIA for LH and FSH, slopes with the greatest maximum binding were obtained at pH 6.4, whereas for the double-antibody method best results were obtained at pH 8.0. Since there is individual variation in the pH of various serum samples, different values will be obtained from samples containing the same amount of hormone. It is sum samples be standardized by the use of buffers with the capacity to keep the pH of the serum constant and that serum standards be dialyzed.

Sixty-one sera from different trimesters of pregnancy were analyzed by two 125-I-NIH-HCG assay systems, employing anti-intact HCG serum and anti theta-HCG serum, respectively. The ratios of the levels measured in the two assay systems changed with the duration of pregnancy. The ratios during trimesters 1,2, and 3 were 2.94, 1.99, and 2.37, respectively. The cross-reactivity of proteohormones other than HCG was tested in both the assay systems. The two assay systems could be comparable in their high degree of specificity. However, the relative affinities of intact HCG and theta-HCG in the two assay systems were observed to be different. It was suggested that the significant differences in the ratios of the levels measured by the tow assay systems might have been infulenced by the occurrence of theta-HCG in serum and that the levels of the subunit must have changed at different stages of pregnancy.

Two radioimmunoassay (RIA) procedures for measuring avian luteinizing hormone (LH) were compared using divided aliquots of plasma samples collected during the ovulatory cycles of turkey and broiler breeder hens in three separate experiments. The RIA systems compared were the homologous turkey LH assay Wentworth et al. (1976) and the homologous chicken LH assay of Follett et al. (1972). Both assays provided the same assessment of circulating LH changes. The preovulatory surge of LH was observed 6 hr before ovulation in turkeys and 4 hr before ovulation in chickens. In addition, both assays detected an increase in chicken LH after ovulation, followed by a gradual decline prior to the next preovulatory surge. The correlation between RIA were highly significant (P less than .001) in each experiment. The crossreaction of purified ostrich LH, FSH, and TSH and of the alpha and beta subunits of turkey LH was assessed in each assay system. Ostrich LH, FSH, and TSH crossreacted in the chicken LH assay in a dose-related manner: ostrich LH showed the highest activity. Only ostrich TSH produced a response in the turkey assay that was dose related. The alpha subunit, but not the beta subunit, of turkey LH was quite potent in the chicken LH RIA, but neither subunit competed as well as the native hormone in the turkey RIA. Thus, the two LH antisera recognize different portions of the LH molecule. The chicken antiserum apparently recognizes a site on the alpha subunit that may be at least partially masked in the FSH and TSH molecules. The turkey antiserum recognizes a site on the native LH molecule that is altered when the subunits are separated.

Monoclonal antibodies were prepared against human follitropin by fusion of the myeloma cell line P3-X63-Ag8-653 with spleen cells of mice immunized with either human follitropin or follitropin bound to an anti-beta hFSH monoclonal antibody. The latter immunization procedure permits shielding of the immunodominant specific site and allows the production of numerous specific monoclonal antibodies to human follitropin which are complementary to the monoclonal antibody used in the immunogen. In this investigation two specific monoclonal antibodies were used in a two site immunoradiometric assay which was highly specific, rapid, with one incubation step and demonstrated a sensitivity level of 0.1 mIU/ml. It was possible to differentiate between prepubertal and adult levels of follitropin and to recognize individuals with hyposecretory states.

Twelve mouse monoclonal antibodies for human luteinising hormone were produced. The affinities varied from 4 X 10(7) to 1 X 10(10) l/mol. The specificity of each antibody was assessed by determining the relative reactivities with luteinising hormone, thyroid stimulating hormone, follicle stimulating hormone and chorionic gonadotrophin. Six antibodies bound to the alpha-subunit as shown by similar reactivity with all hormones, and the remainder to the beta-subunit as shown by specificity for luteinising hormone. This latter group of antibodies cross-reacted only weakly with thyroid stimulating hormone (approximately 10%) and follicle stimulating hormone (approximately 3%). Three of these antibodies also showed low reactivity towards chorionic gonadotrophin (less than 10%), though the others did not (80-300%). The ability of different antibodies to bind simultaneously to luteinising hormone was examined and it was shown that several distinct antigenic determinants existed on both subunits. The characterisation of monoclonal antibody binding sites is discussed in relation to the use of antibodies in two-site immunoradiometric assays.

The possibility of controlling fertility by antibodies inactivating key reproductive hormones has been amply demonstrated by active and passive immunization in primates. Four birth control vaccines directed against human chorionic gonadotropin are currently in early clinical trials. The nature of these vaccines and the underlying principles are described, as are the available results from clinical studies. The alpha- and beta-subunits of human chorionic gonadotropin and the ovine gonadotropins have been cloned by recombinant deoxyribonucleic acid methods. A new breed of vaccines that combines the genes of gonadotropins linked to hepatitis B surface protein has been developed. The next generation of birth control vaccines is likely to be polyvalent and to have the ability to intercept fertility at more than one point. A number of monoclonal antibodies against human sperm have shown the presence of tissue-specific antigens and the possibility of preventing the fertilization of the egg. Inclusion of more than one carrier in the vaccine increases the percentage of high responders and accords immunoprophylactic benefits against more than one disease. Conjugates have also been developed to obtain high titers of antibodies against gonadotropin-releasing hormone with permissible adjuvants. This vaccine may have therapeutic applications in hormone-dependent cancers and precocious puberty.