anti-FSH Antibodies Research Articles

Macro-FSH is a rare cause of inappropriately high FSH concentrations.

Assessment of hormone concentrations can be subjected to laboratory pitfalls. Macro-hormones are hormone-autoantibody complexes which are cleared slowly from circulation and cause a false elevation in hormones' concentrations. Macro-prolactin and macro-thyroid-stimulating hormone (TSH) are most frequently encountered while macro-follicle-stimulating hormone (FSH) has been rarely reported. We describe the case of a 30-year-old woman who had a gynaecological consultation due to failure in achieving pregnancy after 8 months of unprotected intercourse. She had regular menses, did not complain of climacteric symptoms and her medical history was unremarkable. Antral follicle count and anti-mullerian hormone concentrations were normal, and regular ovulation was documented. Unexpectedly, high early follicular phase FSH concentrations were confirmed on two occasions (57 and 51 IU/L), raising the suspicion of primary ovarian insufficiency. After excluding Turner's syndrome and autoimmune oophoritis, a laboratory artifact was hypothesized. Following polyethylene glycol precipitation, FSH levels dropped from 41.1 IU/L to 6.54 IU/L (recovery 16%) and the presence of macro-FSH was concluded. Laboratory interference can lead to misdiagnosis and unnecessary treatments. A laboratory artifact should be suspected when inconsistency exists between clinical presentation and laboratory results. Only five other cases of macro-FSH have been reported to date. Although macro-hormones generally have low biological activity and do not require treatment, the role of anti-FSH antibodies has been hypothesized in primary ovarian insufficiency and in vitro fertilization failure. Hormone quantification is a cornerstone in the diagnostic workup of endocrine disorders, but it can be subjected to laboratory interferences which can lead to unnecessary investigations and inappropriate treatments. A laboratory artifact should be suspected when a discrepancy is observed between clinical presentation and laboratory results, when extremely unusual analyte concentrations are observed and when inconsistent results are obtained by different analytical methods. Macro-hormones are hormone-autoantibody complexes which are cleared slowly from circulation and cause a false elevation in hormone concentrations. Macro-prolactin and macro-TSH are most frequently encountered, while macro-FSH has been rarely reported. Macro-hormones can be detected by polyethylene glycol precipitation, gel filtration chromatography, or by using protein G or protein A columns. Although macro-hormones generally have low biological activity and do not require treatment, the role of anti-FSH antibodies has been hypothesized in primary ovarian insufficiency and in vitro fertilization failure.

An Array SPRi Biosensor for the Determination of Follicle-Stimulating Hormone in Blood Plasma.

Follicle-stimulating hormone (FSH) regulates the development, growth, pubertal maturation and reproductive processes of the human body. The determination of serous FSH concentration is significant as an alternative to testicular biopsy in the case of boys suffering from cryptorchidism after orchidopexy, and as a means of determining the menopausal stage in women. The aim of this investigation is to develop a specific array surface plasmon resonance imaging (SPRi) biosensor for the determination of FSH in body liquids such as blood plasma, obtaining sufficient sensitivity to determine FSH at levels characteristic for that hormone in blood plasma, without any signal enhancement. The biosensor consists of a mouse monoclonal anti-FSH antibody attached to the gold surface of a chip via a cysteamine linker. Its linear response range is from 0.08 mIU mL-1 (LOQ) to 20 mIU mL-1, and well covers most of the range of FSH activities found in blood without dilution. The precision of measurement is between 3.2% and 13.1% for model samples, and between 3.7% and 5.6% for spiked plasma samples. Recoveries are in the range from 94% to 108%. The biosensor has good selectivity, and is validated by comparison with ECLE, with good agreement of the results.

FSH and ApoE4 contribute to Alzheimer’s disease-like pathogenesis via C/EBPβ/δ-secretase in female mice

Alzheimer’s disease (AD) is the most common dementia. It is known that women with one ApoE4 allele display greater risk and earlier onset of AD compared with men. In mice, we previously showed that follicle–stimulating hormone (FSH), a gonadotropin that rises in post–menopausal females, activates its receptor FSHR in the hippocampus, to drive AD–like pathology and cognitive impairment. Here we show in mice that ApoE4 and FSH jointly trigger AD-like pathogenesis by activating C/EBPβ/δ-secretase signaling. ApoE4 and FSH additively activate C/EBPβ/δ-secretase pathway that mediates APP and Tau proteolytic fragmentation, stimulating Aβ and neurofibrillary tangles. Ovariectomy-provoked AD-like pathologies and cognitive defects in female ApoE4-TR mice are ameliorated by anti-FSH antibody treatment. FSH administration facilitates AD-like pathologies in both young male and female ApoE4-TR mice. Furthermore, FSH stimulates AD-like pathologies and cognitive defects in ApoE4-TR mice, but not ApoE3-TR mice. Our findings suggest that in mice, augmented FSH in females with ApoE4 but not ApoE3 genotype increases vulnerability to AD-like process by activating C/EBPβ/δ-secretase signalling.

HLA-B*0702 class-I allele, anti-FSH, anti-LH, and vitamin D3: Potential links with polycystic ovary syndrome in women of Erbil city, Iraq

Polycystic Ovary Syndrome (PCOS) is a widespread endocrine-reproductive-metabolic condition with severe implications for females’ health. The role of four essential parameters on PCOS, including Human Leukocyte Antigens (HLA) represented by the HLA-B*0702 allele, anti-Follicle Stimulating Hormone (anti-FSH) antibodies, anti-Luteinizing Hormone (anti-LH) antibodies, and vitamin D3 was investigated. A total of 100 samples were collected from Kurdish women attended the Maternity Teaching Hospital and some private hospitals/laboratories in Erbil City from October, 2021 to January, 2022. The samples were genotyped using a PCR-based technique with specific sequence primers. The levels of anti-FSH and anti-LH antibodies were determined using an Enzyme-Linked Immunosorbent Assay (ELISA), while vitamin D3 levels were measured by an Electrochemiluminescence (ECL) test on Cobas e411 immunoassay system. The Odds Ratio (OR) of 2.167 at a 95% Confidence Interval (CI) of 0.8167 to 6.330, indicated an essential link between the HLA-B*0702 allele and a risk of PCOS. Anti-FSH and anti-LH antibodies were significantly greater in PCOS patients, notably infertile women, than in healthy controls. A significant positive linear correlation was observed between antibodies against FSH and LH in patients. Most patients and controls had hypovitaminosis D3, with a significant difference (P-value  0.01). The results indicated that the HLA-B*0702 allele is associated with PCOS susceptibility and could be used as an immunogenetic marker. It also supported the idea that anti-FSH and anti-LH antibodies are naturally presence antibodies in PCOS patients instead of signs for autoimmunity. Women suffer from PCOS are also more prone to develop vitamin D3 deficiency.

Study of Anti-Ovarian Antibody, Anti-FSH and Anti-LH Antibodies Along with Their Receptors in Polycystic Ovarian Syndrome

A case control study was conducted to compare sixty females (polycystic ovarian syndrome patients ), thirtywith high LH & FSH hormone level “Group 1 “ G1”, thirty with low LH & FSH hormone level “ Group2 “ G2 “ and thirty healthy control groups. The average age is between (15 – 45 years). The followingparameters were estimated: Luteinizing Hormone (LH), Luteinizing Hormone-Receptor (LH-R), Folliclestimulating hormone (FSH) , Follicle-stimulating hormone-Receptor (FSH-R), anti-LH ,anti-FSH , antiovarian antibody (by ELISA Technique ).Rresults showed that the average age of the patients within the first group1( G1) was 28 year, whilethe average age of the patients in the second group2( G2) was 36 years, also the results showed a highcorrelation between group1 ( G1 ) with (Anti FSH antibodies ) (93%), compared with group2 (G2) that(Anti FSH antibodies ) where it was 0% , as well a high correlation 100% between group1 (G1) with (AntiLH antibodies),compared with group2 (G2) where it was 3% . And anti-ovarian antibodies ,compared withgroup 2 .

Relation of Anti FSH Antibodies and Polycystic Ovarian Syndrome in Women

The aim of the study was to detect the level of anti-follicle stimulating hormone antibody in polycystic women and compared to normal control group. A cross-sectional study was carried out in Erbil city from January 15th of October 2018, to 15th of March 2019. The number of polycystic ovary syndrome (PCOS) women under study was 60 women whose ages were between 15 and 45 years old. These patients admitted to Gynecology and Obstetrics Teaching Hospital. The control group who were matched to the patients studied included 30 individuals. The study showed that there is the significant difference between PCOS women and the control group concerning Anti FSH Antibodies level and the highest mean of Anti FSH Ab level occurred in PCOS women (1.61± 0.14 vs. 0.92 ± 0.04). There was a non-significant relation between each one of LH, FSH, LH/FSH ratio with Anti FSH Ab, in this study. The highest rate of PCOS women had irregular menstrual cycle. The study showed that most PCOS women included in the study had hirsutism. The rate of acne recorded among PCOS women in the study was 48.4 while 51.6 without acne. Conclusion: It was concluded that there was a highly statistically significant relation of Anti FSH Ab with PCOS

Level of Anti-FSH and Anti-LH Antibody in PCOS Women and Comparing it with Normal Control Group

Background: Polycystic ovary syndrome is commonest endocrine disorder of female at reproductive age with estimation of its prevalence in general population was 20-33%Objective: To estimate the levels of Anti-FSH Ab, Anti-LH Ab in polycystic women and comparing it with healthy control group.Patients and methods: This case controls study was carried out in Basrah. In this study blood sample from 29 women with PCOS and 29 healthy control women were collected from outpatient and private gynecological clinics and primary health care centers from different area of Basra from August 2016 to March 2018 for estimation of their serum levels of Anti-FSH Ab, Anti-LH Ab by using enzyme linked immunosorbent assay technique.Results: Serum levels of anti-FSH Ab, anti-LH Ab were elevated in PCOS women.Conclusion: Serum levels of anti-FSH Ab, anti-LH Ab is highly statistically significance in PCOS women than in healthy control group.

The Role of Protamine 1 Gene Polymorphism and Anti FSH Antibody in Infertile Couples

Background: Infertility is a worldwide problem which affects approximately 15% of all couples, and is defined as, inability to conceive after twelve months of contraceptive-free unprotected intercourse. Materials and methods: Optical density of anti follicle stimulation hormone antibody in the serum was detected by enzyme–linked immunosorbent assay technique and from which anti follicle stimulation hormone antibody concentrations were evaluated according to cut-off. One polymorphic sites of Protamine 1 Gene was genotyped on 72 couples suffering from infertility. Another group consist of 20 couples apparently healthy individuals. Genotypes were determined by the polymerase chain restriction fragment length polymorphism (PCR-RFLP) method. Results: Anti FSH antibody result demonstrated differences in antibody values between the patients and the control group with significant statistically, the frequency of the reported SNP was no significantly different compared to normal fertile males, which suggest that such SNP may not serve as a good molecular marker for genetic diagnosis of male infertility. Conclusion: The SNP G197T are completely absent and are not associated with male infertility with aforementioned, therefore these SNPs may not represent as a molecular marker for the diagnosis of genetic cause of male infertility in our studied population.

Assessment of the immunogenicity of gonadotrophins during controlled ovarian stimulation.

ProblemGonadotrophin hormones are used for the controlled ovarian stimulation (COS) as part of the in vitro fertilization techniques. Therapeutic proteins have the potential to induce an unwanted immune response.Method of studyThe presence of anti‐FSH, anti‐LH and anti‐hCG antibodies were determined in patients from two different clinical trials after the repeated administration of hMG or FSH.ResultsIn the first study, 27 subjects were screening for the presence of anti‐FSH antibodies. From the 27 patients, only one patient showed the presence of low levels of antibodies. In a second study, 25 patients were screened for the presence of anti‐FSH, anti‐LH and anti‐hCG antibodies. At the end of the study, no patients showed the presence of antibodies.ConclusionThe results of this study suggest that repeated treatment cycles with FSH or hMG in patients undergoing COS for in vitro fertilization can be safely and effectively applied without concerns for immunogenicity.

Greater amberjack Fsh, Lh, and their receptors: Plasma and mRNA profiles during ovarian development

To understand the endocrine regulation of ovarian development in a multiple spawning fish, the relationship between gonadotropins (Gths; follicle-stimulating hormone [Fsh] and luteinizing hormone [Lh]) and their receptors (Gthrs; Fshr and Lhr) were investigated in greater amberjack (Seriola dumerili). cDNAs encoding the Gth subunits (Fshβ, Lhβ, and glycoprotein α [Gpα]) and Gthrs were cloned. The in vitro reporter gene assay using recombinant hormones revealed that greater amberjack Fshr and Lhr responded strongly to their own ligands. Competitive enzyme-linked immunosorbent assays (ELISAs) were developed for measuring greater amberjack Fsh and Lh. Anti-Fsh and anti-Lh antibodies were raised against recombinant chimeric single-chain Gths consisting of greater amberjack Fshβ (or Lhβ) with rabbit GPα. The validation study showed that the ELISAs were precise (intra- and inter-assay coefficient of variation, <10%) and sensitive (detection limit of 0.2ng/ml for Fsh and 0.8ng/ml for Lh) with low cross-reactivity. A good parallelism between the standard curve and serial dilutions of greater amberjack plasma and pituitary extract were obtained. In female greater amberjack, pituitary fshb, ovarian fshr, and plasma E2 gradually increased during ovarian development, and plasma Fsh significantly increased during the post-spawning period. This suggests that Fsh plays a role throughout ovarian development and during the post-spawning period. Pituitary lhb, ovarian lhr, and plasma Lh were high during the spawning period, suggesting that the synthesis and secretion of Lh, and Lhr expression are upregulated to induce final oocyte maturation and ovulation.

Immunization with FSHβ fusion protein antigen prevents bone loss in a rat ovariectomy-induced osteoporosis model

Osteoporosis, a metabolic bone disease, threatens postmenopausal women globally. Hormone replacement therapy (HTR), especially estrogen replacement therapy (ERT), is used widely in the clinic because it has been generally accepted that postmenopausal osteoporosis is caused by estrogen deficiency. However, hypogonadal α and β estrogen receptor null mice were only mildly osteopenic, and mice with either receptor deleted had normal bone mass, indicating that estrogen may not be the only mediator that induces osteoporosis. Recently, follicle-stimulating hormone (FSH), the serum concentration of which increases from the very beginning of menopause, has been found to play a key role in postmenopausal osteoporosis by promoting osteoclastogenesis. In this article, we confirmed that exogenous FSH can enhance osteoclast differentiation in vitro and that this effect can be neutralized by either an anti-FSH monoclonal antibody or anti-FSH polyclonal sera raised by immunizing animals with a recombinant GST-FSHβ fusion protein antigen. Moreover, immunizing ovariectomized rats with the GST-FSHβ antigen does significantly prevent trabecular bone loss and thereby enhance the bone strength, indicating that a FSH-based vaccine may be a promising therapeutic strategy to slow down bone loss in postmenopausal women.

A 30-Day Preclinical Safety Evaluation Study of Recombinant Human Follicle-Stimulating Hormone in Female Rhesus Monkeys

The objective of this study was to identify potential target organs for toxicity of recombinant human follicle stimulating hormone (r-hFSH) in female rhesus monkeys and to establish a no observed adverse effect level (NOAEL). In all, 24 female rhesus monkeys (Chinese origin, weighing 3.4-5.2 kg, approximately 5 years of age) received repeated subcutaneous (sc) r-hFSH at doses of 10, 60, and 300 IU/kg per d or vehicle once daily for 30 days followed by a 15-day recovery period. Endometrial hyperplasia and dermal edema in the external genitals were found in some animals at 300 IU/kg per d. Pharmacologic-related multiple cystic follicles were found in all r-hFSH-treated groups. A weak, anti-FSH antibody response was detected at the end of treatment in animals administered 60 and 300 IU/kg per d. These results indicate that the primary effects of r-hFSH in female rhesus monkeys were related to its pharmacological activity on the reproductive system. The NOAEL was considered to be 60 IU/kg per d.

Effect of recombinant gonadotropins on embryo quality in superovulated rabbit does and immune response after repeated treatments

This study aimed first to evaluate the effect of recombinant human FSH (rhFSH) with and without recombinant human LH (rhLH) on fresh and frozen-thawed embryo development and also to analyze the immune response of rabbit does (Oryctolagus cuniculus) subjected to repeated rhFSH treatments. Nulliparous New Zealand White does were used. In Experiment 1, 120 does were superovulated with 25 IU rhFSH alone or in combination with 5% or 10% rhLH (1.25 IU or 2.50 IU rhLH). A total of 1116 embryos at the compacted morula stage were cultured at 38.5 °C, 5% CO 2, and saturated humidity for 48 h. The embryo development to hatching blastocyst was significantly lower for the group with 10% rhLH versus that of the control group (65.6 vs. 79.5 for rhFSH + 10% rhLH vs. control, respectively). However, no significant difference was found in development to hatching blastocyst for the control, rhFSH alone, and rhFSH + 5% rhLH groups. The developmental potential of frozen-thawed embryos obtained from all groups was similar, with an 83.5% in vitro development rate until the expanded blastocyst stage. To detect anti-FSH antibodies, in Experiment 2, does were subject to four superovulation treatments. The hormone administration had a significant effect on immune response in the superovulation group after two treatments (0.14 ± 0.074 and 0.15 ± 0.076 vs. 0.46 ± 0.078 and 0.50 ± 0.078 optical density for the first, second, third, and forth cycles, respectively). Nevertheless, none of the treated does had an immune response in both the first and second treatments; on the contrary, a significant increase in the antibody levels was observed in these females at the moment of the third and fourth superovulation treatments. In conclusion, rhFSH superovulation treatments increase the reproductive potential of rabbit does.

Follicle-stimulating hormone increases cholangiocyte proliferation by an autocrine mechanism via cAMP-dependent phosphorylation of ERK1/2 and Elk-1

Sex hormones regulate cholangiocyte hyperplasia in bile duct-ligated (BDL) rats. We studied whether follicle-stimulating hormone (FSH) regulates cholangiocyte proliferation. FSH receptor (FSHR) and FSH expression was evaluated in liver sections, purified cholangiocytes, and cholangiocyte cultures (NRICC). In vivo, normal female and male rats were treated with FSH or immediately after BDL with antide (a gonadotropin-releasing hormone antagonist blocking FSH secretion) or a neutralizing FSH antibody for 1 wk. We evaluated 1) cholangiocyte proliferation in sections and cholangiocytes and 2) changes in secretin-stimulated cAMP (functional index of cholangiocyte growth) levels, and ERK1/2 and Elk-1 phosphorylation. NRICC were stimulated with FSH before evaluation of proliferation, cAMP/IP(3) levels, and ERK1/2 and Elk-1 phosphorylation. To determine whether FSH regulates cholangiocyte proliferation by an autocrine mechanism, we evaluated the effects of 1) cholangiocyte supernatant (containing FSH) on NRICC proliferation and 2) FSH silencing in NRICC before measuring proliferation and ERK1/2 and Elk-1 phosphorylation. Cholangiocytes and NRICC express FSHR and FSH and secrete FSH. In vivo administration of FSH to normal rats increased, whereas administration of antide and anti-FSH antibody to BDL rats decreased 1) ductal mass and 2) secretin-stimulated cAMP levels, proliferation, and ERK1/2 and Elk-1 phosphorylation in cholangiocytes compared with controls. In NRICC, FSH increased cholangiocyte proliferation, cAMP levels, and ERK1/2 and Elk-1 phosphorylation. The supernatant of cholangiocytes increased NRICC proliferation, inhibited by preincubation with anti-FSH antibody. Silencing of FSH gene decreases cholangiocyte proliferation and ERK1/2 and Elk-1 phosphorylation. Modulation of cholangiocyte FSH expression may be important for the management of cholangiopathies.

Follicle-stimulating hormone autoantibody is involved in idiopathic spermatogenic dysfunction

To detect the anti-follicle-stimulating hormone (FSH) antibody in idiopathic infertile patients and fertile subjects in order to determine the role of this antibody in patients with spermatogenic dysfunction. The anti-FSH antibody in serum was detected by an enzyme-linked immunosorbent assay (ELISA). The functional and structural integrity of the sperm membrane was evaluated with hypo-osmotic swelling (HOS) test and the ultrastructure of the spermatozoa was investigated by transmission electron microscopy (TEM). The extent of positive FSH antibody in the patients with oligozoospermia and/or asthenozoospermia was significantly higher than that in the fertile subjects and infertile patients with normal sperm concentration and motility, but it was significantly lower than that in the patients with azoospermia. The extent of anti-FSH antibody in the patients with azoospermia was significantly greater than that in patients with oligospermia and/or asthenospermia, infertile people with normal sperm density and motility and fertile people. The hypo-osmotic swelling test showed that the percentage of HOS-positive spermatozoa (swollen) was 45.1 mu 3.5% in the FSH antibody-positive group and 59.1% micro 6.2% in the FSH antibody-negative control group. The percentage of functional membrane damage to spermatozoa was significantly higher in the anti-FSH antibody-positive group than in the control group. TEM showed that the outer acrosomal membrane was located far from the nucleus, and detachment of the acrosome was found in the FSH autoantibody-positive group. These data suggest that the presence of anti-FSH antibody is strongly correlated with the sperm quantity and quality in idiopathic male infertility. Anti-FSH antibody may be an important factor causing spermatogenic dysfunction and infertility.

Pathways involved in testicular germ cell apoptosis in immature rats after FSH suppression

FSH is a key regulator of testis function, required for the establishment of full complements of Sertoli and germ cells during postnatal testis development and for the maintenance of spermatogenesis in the adult. FSH plays an important role in germ cell survival rather than proliferation, in the window between 14 and 18 days of testicular development, which coincides with the cessation of Sertoli cell proliferation and the onset of germ cell meiosis during the first wave of spermatogenesis. This study aimed to identify the pathway(s) of apoptosis regulated by changes in FSH levels in 14 - to 18-day-old rats, using a model of in vivo FSH suppression by passive immunoneutralization with a rat anti-FSH antibody. Apoptotic pathways were identified by immunohistochemistry using pathway-specific proteins as markers of the intrinsic (activated caspase 9) and extrinsic (activated caspase 8) pathways, followed by quantification of cell numbers using stereological techniques. In addition, RT-PCR was used to assess the expression of pathway-specific genes. We previously reported a 2.5-fold increase in spermatogonial apoptosis in these samples after 4 days of FSH suppression, and now show that this increase correlates with a 9.8-fold (P<0.001) increase in the frequency of caspase 9-positive spermatogonia in the absence of caspase 8 immunoreactivity. By contrast, spermatocytes exhibited both increased caspase 9 (7.5-fold; P<0.001) and caspase 8 (5.7 fold; P<0.001) immunoreactivities after 4 days of FSH suppression. No significant change in the transcription levels of candidate genes required for either pathway was detected. This study demonstrates that, in the seminiferous tubules, FSH suppression induces spermatogonial apoptosis predominantly via the intrinsic pathway, while spermatocyte apoptosis occurs via both the intrinsic and extrinsic pathways.

Anti-FSH antibodies associate with poor outcome of ovarian stimulation in IVF

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 (≤3 oocytes) (adjusted odds ratio for IgG = 6.95, P = 0.005 and IgA = 3.60, P = 0.039). FF anti-FSH IgG and IgA were positively associated with serum anti-FSH concentrations and FSH concentration in FF. Additionally, FF anti-FSH IgG increased with follicle growth (linear regression coefficient = 0.02, P = 0.022). Collectively, these data suggest that serum anti-FSH antibodies are associated with poor ovarian response to FSH stimulation in IVF, with anti-FSH IgA and IgG potentially exerting a local FSH antagonizing effect in maturing follicles.

Putative Predictors of Antibodies Against Follicle‐Stimulating Hormone in Female Infertility: A Study Based on In Vitro Fertilization Patients

We have previously demonstrated the presence of naturally occurring antibodies against follicle-stimulating hormone (FSH) in patients with endometriosis and polycystic ovary syndrome (PCOS). Here, we investigated the parameters associated with anti-FSH antibodies in in vitro fertilization (IVF) patients. The following parameters were studied in 135 patients: peripheral FSH levels, FSH beta-subunit gene (FSHB) haplotypes, history of previous IVF, and susceptibility to autoimmune reactions in general [seven common autoantibodies (against nuclear antigens on human and rodent substrates, smooth muscle, gastric parietal cells, beta2-glycoprotein I, cardiolipin, and thyroid peroxidase) and HLA-DQB1 alleles]. Although the anti-FSH levels were higher in patients when compared with controls, those higher levels were not associated with FSHB haplotypes. The anti-FSH IgM associated with (i) the levels of FSH in women with male and tubal factor infertility; (ii) the history of IVF in patients with PCOS, endometriosis, and unexplained infertility; and (iii) the production of common autoantibodies among all IVF patients. The anti-FSH IgA associated with HLA-DQB1*03. The anti-FSH IgG correlated with the values of anti-FSH IgA and IgM. Anti-FSH may be naturally occurring antibodies associated with peripheral FSH concentrations, but increased in infertile women with dysregulation of immune reactions and repeatedly performed IVF.

P-518: Effect of LBFS0101, a new recombinant follicle-stimulating hormone, in human in vitro fertilization and embryo transfer program: A prospective and randomized clinical study

The purpose of this study was to analyze the efficacy and tolerability of follitropin-heterodimeric alpha-beta subunit mixture (LBFS0101; LG Life Sciences Ltd., Seoul, South Korea), new recombinant human FSH (r-hFSH) preparations, for superovulation in patients undergoing IVF-ET. Randomized, parallel-group, open, multi-center clinical study. One hundred and three infertile women undergoing IVF-ET in 2005 were entered into the study under approval of the Institutional Reviewer Board. After down-regulation with buserelin acetate, patients were randomized to receive LBFS0101 (n=52) or Gonal-F (Serono Inc., Geneva, Switzerland, n=51). r-hFSH was administrated at 150-300 IU/d for 3∼5 days, and then dosages were adjusted according to the ovarian response. Ovarian stimulation characteristics, ovum retrieval, and IVF/ICSI outcomes were compared. Total of 15 cycles were cancelled, 88 cycles (LBFS0101 = 47 cycles; Gonal F = 41 cycles) were studied. There were no statistically significant differences in any clinical profiles of patients between the two preparations. Also, cumulative dose of r-hFSH (2371.3 ± 728.4 mIU for LBFS0101 vs.2409.8 ± 769.4 mIU for Gonal F), duration of r-hFSH treatment (9.3 ± 2.0 days vs. 9.3 ± 1.8 days), number of follicles of ≥ 14 mm on the of hCG injection (9.3 ± 4.6 days vs. 10.6 ± 5.8), and number of retrieved oocytes (14.2 ± 9.1 vs. 14.4 ± 8.8) were not different. Clinical pregnancy rate and implantation rates were similar (44.7% (21/47), 22.1% (38/169) for LBFS0101 and 56.1% (23/41), 29.5% (41/139) for Gonal F). Anti-FSH antibody was not detected in all samples. The results of this clinical study indicate that LBFS0101 may be suitable for use in ovarian stimulation for human IVF-ET program.

Both Testosterone and Follicle-Stimulating Hormone Independently Inhibit Spermatogonial Differentiation in Irradiated Rats

Simultaneous suppression of both testosterone and FSH with GnRH antagonists (GnRH-ant) reverses the radiation-induced block in spermatogonial differentiation in F1 hybrids of Lewis and Brown-Norway rats. Although addition of exogenous testosterone restores the block, it also raises FSH, and hence it had not been possible to conclusively determine which hormone was inhibiting spermatogonial differentiation. In the present study, we establish the relative roles of testosterone and FSH in this inhibition using three different approaches. The first approach involved the treatment of irradiated rats, in which differentiation was stimulated by GnRH-ant plus flutamide, with FSH for 2 wk; the FSH reduced the percentage of tubules that were differentiated (TDI) by about 2-fold, indicating that FSH does have an inhibitory role. The second approach involved treatment of irradiated, hypophysectomized rats with exogenous testosterone for 10 wk; testosterone also reduced the TDI, demonstrating that testosterone had a definite inhibitory effect, independent of pituitary hormones. Furthermore, in this protocol we showed that TDI in the hypophysectomized testosterone-treated group, which had higher intratesticular testosterone levels but lacked FSH, was slightly higher than the TDI in a GnRH-antagonist-testosterone-treated group of irradiated rats, which had normal physiological levels of FSH; this result supports a role for endogenous FSH in suppressing spermatogonial differentiation in the latter group. The third approach involved injection of an active anti-FSH antibody for 10 d in untreated, GnRH-ant plus flutamide-treated, or GnRH-ant plus testosterone-treated irradiated rats. This was not sufficient to increase the TDI. However, flutamide given in a similar treatment schedule did increase the TDI in GnRH-ant plus testosterone-treated rats. We conclude that both testosterone and FSH individually inhibit spermatogonial differentiation after irradiation, but testosterone is a more highly potent inhibitor than is FSH.