Presence of PSA antibodies in seminal plasma of infertile men.

Immunoinfertility due to antisperm antibodies and semen hyperviscosity are among major causes of male infertility. Although the modulation of prostate-specific antigen (PSA) has been investigated in prostate abnormalities, its role and the effect of its dysfunction in male fertility/infertility have not been extensively examined. The present study was conducted to examine the presence of PSA antibodies locally in the seminal plasma of men having immunoinfertility and semen hyperviscosity. Seminal plasma samples from immunoinfertile men (n=25), men with hyperviscous semen (n=25), and normal men (n=24) were collected and analyzed for immunoreactivity with PSA in ELISA and Western blot. In the immunoinfertile group, seminal plasma from 20% of men reacted positively with PSA. In the hyperviscous group, seminal plasma from 28% of men reacted positively with PSA. None (0%) of the seminal plasma from the normal group showed immunoreactivity to PSA. This is the first study ever to indicate the presence of PSA antibodies in semen of men having immunoinfertility or hyperviscosity. These findings may have clinical significance in the specific diagnosis and treatment of infertility in men and contraceptive vaccine development.

Immunocontraception for Animals: Current Status and Future Perspective.

An alternative to surgical sterilization for fertility control of animals (wild, zoo, farm, and domestic) is needed to prevent problems related to overpopulation, including culling and relocation. A PubMed and Google Scholar database search was conducted using the keywords 'contraceptive vaccine animals,' 'immunocontraception animals,' 'non-surgical sterilization animals,' 'PZP vaccine,' and 'GnRH vaccine.' The searches from 1972 to 2015 yielded over 1500 publications. These articles were read, and 375 were selected for detailed analysis. Articles referenced in these publications were also thoroughly examined. PZP and GnRH contraceptive vaccines (CVs) have been extensively investigated for fertility control of wild, zoo, farm, and domestic animal populations. Both vaccines have shown tremendous success with PZP vaccines taking the lead. Novel technologies and targets are being developed to improve existing vaccines and generate second-generation CVs. Single-shot vaccines, which can be delivered remotely, will greatly advance the field of immunocontraception for animal use with potential human application.

Curcumin: a novel non-steroidal contraceptive with antimicrobial properties.

Women face unique pathologies and challenges related to the female genital tract (FGT), including vaginal infections and gynecologic cancers. Vaginal infections faced by women include bacterial vaginosis (BV), vulvovaginal candidiasis (VC), and sexually transmitted infections (STIs). Curcumin, a component of the dietary spice turmeric, has immense biological properties, including antioxidant, anti-inflammatory/immunomodulatory, and anticancer effects. It has no side effects and is well-tolerated, making it an ideal treatment modality highly desired by women. Recently, our laboratory showed, for the first time ever, that curcumin exhibits a spermicidal and broad-spectrum microbicidal activity against several species of bacteria and yeast involved in vaginal infections. Thus, it could provide a novel, non-steroidal contraceptive having both spermicidal and microbicidal properties and can be panacea in women for treatment of several FGT pathologies, including gynecologic cancers.

The Effect of Curcumin on Intracellular pH (pHi), Membrane Hyperpolarization and Sperm Motility.

BACKGROUND: Curcumin has shown to affect sperm motility and function in vitro and fertility in vivo. The molecular mechanism(s) by which curcumin affects sperm motility has not been delineated. Since modulation of intracellular pH (pHi) and plasma membrane polarization is involved in sperm motility, the present study was conducted to investigate the effect of curcumin on these sperm (human and murine) parameters. METHODS: The effect of curcumin on sperm forward motility was examined by counting percentages of forward moving sperm. The effect of curcumin on intracellular pH (pHi) was measured by the fluorescent pH indicator 2,7-bicarboxyethyl-5,6-carboxyfluorescein-acetoxymethyl ester (BCECF-AM). The effect of curcumin on plasma membrane polarization was examined using the fluorescence sensitive dye bis (1,3-dibarbituric acid)-trimethine oxanol [DiBAC4(3)]. RESULTS: Curcumin caused a concentration-dependent (p<0.05) decrease in forward motility of both human and mouse sperm. It also caused a concentration-dependent decrease in intracellular pH (pHi) in both human and mouse sperm. Curcumin induced significant (p<0.05) hyperpolarization of the plasma membrane in both human and mouse sperm. CONCLUSION: These findings indicate that curcumin inhibits sperm forward motility by intracellular acidification and hyperpolarization of sperm plasma membrane. This is the first study to our knowledge which examined the effect of curcumin on sperm pHi and membrane polarization that affect sperm forward motility. These exciting findings will have application in deciphering the signal transduction pathway involved in sperm motility and function and in development of a novel non-steroidal contraceptive for infertility.

Vaccine for human contraception targeting sperm Izumo protein and YLP12 dodecamer peptide.

There is an urgent need to develop a better method of contraception which is non-steroidal and reversible to control world population explosion and unintended pregnancies. Contraceptive vaccines (CV), especially targeting sperm-specific proteins, can provide an ideal contraceptive modality. Sperm-specific proteins can induce an immune response in women as well as men, thus can be used for CV development in both sexes. In this article, we will review two sperm-specific proteins, namely Izumo protein and YLP12 dodecamer peptide. Gene-knockout studies indicate that Izumo protein is essential for sperm-egg membrane fusion. Vaccination with Izumo protein or its cDNA causes a significant reduction in fertility of female mice. The antibodies to human Izumo inhibit human sperm penetration assay. Recently, our laboratory found that a significant percentage of infertile women have antibodies to Izumo protein. The second sperm-specific protein is YLP12 , a peptide mimetic sequence present on human sperm involved in recognition and binding to the human oocyte zona pellucida. Vaccination with YLP12 or its cDNA causes long-term, reversible contraception, without side effects, in female mice. Infertile, but not fertile, men and women have antibodies to YLP12 peptide. Our laboratory has isolated, cloned, and sequenced cDNA encoding human single chain variable fragment (scFv) antibody from infertile men which reacts with YLP12 peptide. The human YLP12 scFv antibody may provide a novel passive immunocontraceptive, the first of its kind. In conclusion, sperm-specific Izumo protein and YLP12 peptide can provide exciting candidates for antisperm CV development.

Polycystic ovary syndrome: current status and future perspective.

Polycystic ovary syndrome (PCOS) is a widespread reproductive disorder that encompasses many associated health conditions and has an impact on various metabolic processes. PCOS is depicted by hyperandrogenism, polycystic ovaries, and anovulation. It increases the risk of insulin resistance (IR), type 2 diabetes, obesity, and cardiovascular disease. The etiology of the disease remains unclear, and the subjective phenotype makes a united diagnosis difficult among physicians. It seems to be a familial genetic syndrome caused by a combination of environmental and genetic factors. It can be linked with metabolic disorders in first-degree family members. PCOS is the cause of up to 30% of infertility in couples seeking treatment. Currently, there is no cure for PCOS. Despite the growing incidence of this syndrome, limited research has been done that encompasses the entirety of PCOS spectrum. In this review, the current status and possible future perspective will be discussed.

Recent progress toward development of vaccines against conception.

Although there are several methods of contraception available, the population explosion and unintended pregnancies continue to cause major health issues and constrain resources worldwide. A better method of contraception that is acceptable in developed as well as developing countries is urgently needed. Contraceptive vaccines (CVs) can fulfil most of the properties of an ideal contraceptive. The development of CVs is an exciting proposition and has drawn significant attention lately. The purpose of this article is to review the present status and future perspectives of CVs for human use, and for control of farm, feral, stray and domestic animal populations.

Antibodies to prostate-specific antigen in immunoinfertile women and men.

Immunoinfertility is caused by antisperm antibodies (ASA) present in the female or male partner of an infertile couple. PSA is a serine protease produced primarily by the prostate gland and is present in human semen. Although the modulation of PSA has been investigated in prostate abnormalities, its role and effect of its dysfunction on human fertility has not been examined. The present study was conducted to examine the presence and incidence of antibodies to PSA in sera of immunoinfertile women and men. The presence of iso- and autoantibodies in the sera of immunoinfertile patients was examined using the well-characterized purified human PSA and seminal plasma from normal fertile men in ELISA. Sera were collected from infertile women (n=25) and infertile men (n=19) who were positive for ASA. Sera from ASA-negative fertile women (n=24) and fertile men (n=19) served as controls. In women, sera from 40% of the immunoinfertile women showed positive reactivity with PSA, and 20% reacted positively with seminal plasma. None of the sera from fertile women reacted positively with PSA or seminal plasma. In men, sera from 16% of the immunoinfertile men showed positive reactivity with PSA, and none (0%) with seminal plasma. None of the sera from fertile men reacted positively with PSA or seminal plasma. This is the first study to examine the presence and incidence of PSA antibodies in immunoinfertile women and men. The findings may have clinical application in the specific diagnosis and treatment of immunoinfertility in humans.

Presence and incidence of izumo antibodies in sera of immunoinfertile women and men.

PROBLEM: Sperm antigens that are relevant to fertilization and fertility, and are involved in human immunoinfertility, are being investigated in several laboratories. Izumo is a sperm-specific protein that has shown to be essential for sperm-egg fusion. The present study was conducted to examine the expression of Izumo protein in human sperm. Also, the presence and incidence of Izumo antibodies in sera of immunoinfertile women and men were investigated. METHOD OF STUDY: The expression of Izumo protein in human sperm was examined by the Western blot procedure. Sera from immunoinfertile women (n = 25) and fertile women (n = 23), as well as sera from immunoinfertile men (n = 20) and fertile men (n = 15), were collected and analyzed for immunoreactivity with Izumo peptides in the enzyme-linked immunosorbent assay (ELISA). Three epitopes of human Izumo, namely one epitope of Izumo1, and two epitopes of Izumo2 [Izumo2 (K-15), and Izumo2 (T-17)], respectively, were selected for the present study. RESULTS: In the Western blot procedure, the Izumo1 antibody recognized a specific protein band of ~39 kDa in human sperm extract. In ELISA, 56% of the immunoinfertile female sera reacted positively (≧3 S.D. units) with Izumo1, 40% with Izumo2 (K-15), and 20% with Izumo2 (T-17) peptide. None of the sera (0%) from fertile women reacted positively (≦3 S.D. units) with any of the three Izumo peptides. None of the sera from immunoinfertile (0%) or fertile men (0%) reacted positively with any of the three Izumo peptides. CONCLUSION: Our findings indicate that human sperm expresses Izumo protein, and the immunoinfertile female sera have circulating isoantibodies against this protein. This is the first study to our knowledge, which examined the presence and incidence of Izumo antibodies in female and male immunoinfertility. These findings may find clinical applications in specific diagnosis and treatment of infertility, and contraceptive vaccine development.

Female genital tract immunity: distinct immunological challenges for vaccine development.

The population explosion and unintended pregnancies, sexually transmitted diseases including human immunodeficiency virus, and cervical cancer, are major challenges to health worldwide. Their prevention might be achieved through vaccination-based approaches to activate specific immunity against pathogen- or fertility-associated antigens in the female genital tract (FGT). This article aims to review methodologies for enhancing adaptive immunity in the FGT to maximize the response to vaccination. Most components of the adaptive and innate mucosal immune system are present in the FGT and several features are common with the nasopharynx/bronchial and gastrointestinal tracts. In contrast to other mucosal sites, the FGT has minimal local lymphoid tissue. Other sites primarily produce IgA and IgM while in the FGT, especially the vaginocervix, IgG is the predominant immunoglobulin secreted. In rodents, data exist to substantiate a common mucosal immune system interconnecting the nasal/bronchial, gastrointestinal, and female genital tracts. The intranasal route seems the most efficacious to induce an immunity in the FGT especially when combined with a systemic or parenteral route. In humans, for induction of secretory IgA and IgG antibodies in the FGT, immunization by the nasal or the vaginal route is effective. In vaginal immunization, a strong and consistent antibody response is best achieved following vaccination during the follicular phase of the menstrual cycle. Antibodies administered systemically percolate into the FGT and can provide immunoprotection against target molecules or cells. Thus, as well as active immunization using selected routes, the passive immunization approach may provide a viable alternative to vaccinology for future development.

Prophylactic vaccines for prevention of prostate cancer.

Prostate cancer is the most prevalent cancer and the second leading cause of cancer death in men. There are various modalities for treatment of prostate cancer. Immunotherapy with several vaccines and antibodies has also been successfully used with positive clinical outcome in prostate cancer patients. The majority of these vaccines are palliative and have been employed when a person is already diagnosed with prostate cancer. The aim of this article is to review various vaccines that have been examined for immunoprophylactic prevention of initiation/development/metastasis of prostate cancer. The Pubmed database and Google Scholar search identified 26 articles on various vaccines that have been investigated for prophylactic prevention of cancer development. These vaccines targeted prostate-specific/restricted antigens (PSA/PSMA/PSCA), oncoproteins (GRP/MUC family, erbB2/HER-2/neu), whole tumor cell antigens, prostate regulating hormones (GnRH/testosterone), and various cytokines and immune modulators. The data indicates that the development of immunoprophylactic vaccines for prostate cancer is an exciting proposition, which can translate into a viable reality for clinical application in humans.

Birth control vaccine targeting leukemia inhibitory factor.

The population explosion and unintended pregnancies resulting in elective abortions continue to impose major public health issues. This calls for a better method of contraception. Immunocontraception has been proposed as a valuable alternative that can fulfill most, if not all, of the properties of an ideal contraceptive. There are several targets that are being explored for contraceptive vaccine development. Leukemia inhibitory factor (LIF), a member of interleukin-6 family, is required for embryo development and successful blastocyst implantation in several mammalian species. The present study was conducted to examine if LIF can be a target for the development of a birth control vaccine. Three sequences from LIF and two sequences from LIF-receptor (LIF-R) that span the regions involved in ligand-receptor binding were delineated, and peptides were synthesized based upon these sequences. Antibodies raised against these five peptides reduced LIF bioactivity in an in vitro culture assay using BA/F3 mLIF-R-mpg130 cells. Vaccines were prepared by conjugating these peptides to various carrier proteins. Immunization of female mice with these peptide vaccines induced a long-lasting, circulating as well as local antibody response in various parts of the genital tract, and resulted in a significant (P ≤ 0.05) inhibition in fertility in all the three trials; the LIF-R peptide vaccines proved to be a better vaccine target. The data indicate that LIF/LIF-R is an excellent target for the development of a birth control vaccine. This is the first study, to our knowledge, that examined LIF/LIF-R as a target for immunocontraception. The findings of this study can be easily translated to humans since LIF/LIF-R is also important for implantation and pregnancy in women.

Antisperm contraceptive vaccines: where we are and where we are going?

This is a review of current status and future perspectives on the development of antisperm contraceptive vaccines (CV) and immunocontraceptives. The development of antisperm CV is an exciting proposition. There is a strong rationale and recent data indicating that this proposition can translate into reality. The search for novel sperm-specific antigens/genes, that can be used for CV, continues using various recent developing technologies. Various approaches of proteomics, genomics, reproductive biology, mucosal immunity and vaccinology and several novel technologies such as gene knockout technology, phage display technology, antibody engineering, differential display technique, subtractive hybridization, and hybridoma technology are being used to delineate sperm-specific antigens and construct CV. Various sperm antigens/genes have been delineated, cloned, and sequenced from various laboratories. Vaccination with these sperm antigens (recombinant/synthetic peptide/DNA) causes a reversible contraceptive effect in females and males of various animal species, by inducing a systemic and local antisperm antibody response. The efficacy is enhanced by combination vaccination, including peptides based on various sperm antigens. Several human novel scFv antibodies with unique complementarity-determining regions (CDRs), that react with specific well-defined fertility-related sperm antigens, have been synthesized. These human infertility-related antibodies may find application in the development of novel immunocontraceptives. Besides finding the novel sperm antigens, the present and future focus is on enhancing the immunogenicity, bioefficacy, and on obliterating the inter-individual variability of the immune response, and proceeding for primate and human clinical trials. Multi-epitope vaccines combining sperm proteins involved in various steps of fertilization cascade have been found to enhance the immunogenicity and bioefficacy of the contraceptive effect. The in vitro synthesis of infertility-related human scFv antibodies may provide unique once-a-month immunocontraceptives, the first of its kind, for human use. The multi-epitope CV and preformed engineered human antibodies of defined specificity may obliterate the concern related to inter-individual variability of the immune response.

Contraceptive vaccines targeting factors involved in establishment of pregnancy.

Current methods of contraception lack specificity and are accompanied with serious side effects. A more specific method of contraception is needed. Contraceptive vaccines can provide most, if not all, the desired characteristics of an ideal contraceptive. This article reviews several factors involved in the establishment of pregnancy, focusing on those that are essential for successful implantation. Factors that are both essential and pregnancy-specific can provide potential targets for contraception. Using database search, 76 factors (cytokines/chemokines/growth factors/others) were identified that are involved in various steps of the establishment of pregnancy. Among these factors, three, namely chorionic gonadotropin (CG), leukemia inhibitory factor (LIF), and pre-implantation factor (PIF), are found to be unique and exciting molecules. Human CG is a well-known pregnancy-specific protein that has undergone phase I and phase II clinical trials, in women, as a contraceptive vaccine with encouraging results. LIF and PIF are pregnancy-specific and essential for successful implantation. These molecules are intriguing and may provide viable targets for immunocontraception. A multiepitope vaccine combining factors/antigens involved in various steps of the fertilization cascade and pregnancy establishment may provide a highly immunogenic and efficacious modality for contraception in humans.

Can curcumin provide an ideal contraceptive?

The population explosion, unintended pregnancies, sexually transmitted diseases, and cancer (cervical and breast) continue to cause major public health issues worldwide. Curcumin, diferuloyl methane, the yellow pigment component of the curry spice turmeric (Curcuma longa), has immense biological effects and has recently drawn considerable attention. Curcumin has antibacterial, antiviral, antiinflammatory, and anticancer properties. It has shown a lack of toxicity in animals and human clinical trials. Yet, its effect on reproduction has not been examined. The present study was conducted to examine if curcumin affects sperm function in vitro and fertility in vivo. Sperm (human and murine) were collected and incubated with curcumin to examine the effect on motility, capacitation/acrosome reaction, and in vitro fertilization. The effect on in vivo fertility using the mouse model was also examined. Incubation of sperm with curcumin caused a concentration-dependent decrease in sperm forward motility, capacitation/acrosome reaction, and murine fertilization in vitro. At higher concentrations, there was a complete block of sperm motility and function within 5-15 min. Administration of curcumin, especially intravaginally, caused a significant (P<0.001) reduction in fertility. The antifertility effect of curcumin was reversible. This is the first study to report the inhibitory effect of curcumin on sperm function, fertilization, and fertility. The findings suggest that curcumin may constitute a double-edged sword to block conception, infection, and cancer, thus providing an ideal contraceptive.

Male-biased genes of Drosophila melanogaster that are conserved in mammalian testis.

Male-biased genes have drawn considerable attention due to their relatively rapid rates of interspecies diversification. These genes are often involved in processes related to reproductive behavior, sexual competition, and gametogenesis. Despite this trend toward rapid evolutionary change, many core molecular pathways underlying spermatogenesis are conserved between Drosophila melanogaster and several mammalian species. Using BLAST search in the database, 22 testis-specific mammalian orthologues of 174 known D. melanogaster male-biased proteins were identified. They are related to a variety of molecular processes. Several also showed association with human male-factor infertility. These genes/proteins may find applications in the diagnosis and treatment of male infertility and the development of novel contraceptives.

Gene knockouts that affect female fertility: novel targets for contraception.

The knockout gene technology is an excellent tool to investigate gene function. In 2005, we reviewed 83 gene knockouts that were reported to affect female fertility. This article is an update of the previous review published in early 2005. It describes genes discovered (2004-2010) since the last review that affect female fertility. Using the database search in the Pubmed and Google Scholar search engines, 67 new genes were discovered using knockout technology that have been reported to affect female fertility. These genes were grouped into three main categories based upon the aspect of female reproductive biology that is affected by their knockout. Some of these genes may provide novel targets for developing better methods for contraception and specific diagnosis and treatment of female infertility.

Identification of human sperm proteins that interact with human zona pellucida3 (ZP3) using yeast two-hybrid system.

Sperm proteins that interact with zona pellucida 3 (ZP3) have not been clearly identified in humans. In the present study, the yeast two-hybrid (Y2H) system was used to identify human sperm proteins that interact with human ZP3. Human ZP3 cDNA was cloned into pAS2-1 yeast vector and used as bait to find reactive proteins in the human testis cDNA library. Six specific clones were obtained that were further confirmed for interaction using the mammalian two-hybrid system. These six clones showed homologies with several proteins in the GenBank database. Of these, the strongest ZP3-interacting protein, that shows 97% homology with ubiquitin associated protein-2 like (UBAP2L), was tested in the hemizona assay. UBAP2L antibodies significantly (p<0.001) inhibited human sperm-zona binding in this assay. We conclude that the Y2H system is a useful strategy for identifying novel genes encoding proteins that interact with ZP proteins. To our knowledge, this is the first study using the Y2H system to identify sperm proteins that interact with human oocyte ZP3. Novel proteins identified using this system may find applications in elucidating the fertilization cascade, development of a new generation of non-steroidal contraceptives, and specific diagnosis and treatment of human infertility.

Novel biomarkers and therapeutic targets for prostate cancer.

Prostate cancer is the most prevalent cancer in the Western male population and the second leading cause of cancer death in men, affecting over 10 million individuals. Present approaches to control the cancer mortality have focused on the detection of the cancer at early stages when it is still locally confined and may be curable. Identification of the prostate-specific antigen (PSA) has facilitated the early diagnosis of prostate cancer. However, PSA has limited specificity and sensitivity in appropriately detecting early stages of abnormal prostate growth. PSA levels fail to differentiate between indolent and aggressive cancers, do not correlate with tumor size, and cross-react with other serine proteases namely, glandular kallikreins 1 and 2. Besides cancer, its levels also increase in men with benign prostatic hyperplasia (BPH), prostatitis, and other non-malignancies. Additional prostate-specific genes and metabolites need to be identified to provide a better understanding of the molecular mechanisms of prostate physiology and pathophysiology. Novel markers for the diagnosis and development of new treatment modalities are urgently needed.