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Lactate dehydrogenase--X--separation of human spermatozoa isoenzymes by extraction with detergents.

Proteins with known enzymatic properties are attractive molecules as possible agents for immunological contraception. Therefore a new technique for the selective isolation of LDH-X from human spermatozoa is described. Applying the detergents hyamine 2389 and triton X-100 LDH-X is extracted from the sperm surface membrane without the other LDH-isoenzymes. Ultrasonification of spermatozoa yields all LDH-isoenzymes together. After cell disruption the separation of LDH-X and the isoenzymes is performed by discelectrophoresis. With lactate as substrate tetrazolium staining identifies all LDH-enzymes. The technique represents an attractive method for the selective isolation of LDH-X and opens further purification steps.

Detergents↗

Molecular dissection of human testicular germ cell differentiation with monoclonal antibodies.

Monoclonal antibodies (MoAbs) were raised by immunising mice with nucleated cells other than sperm (NCOS) obtained from semen of an oligospermic donor. These MoAbs recognised different germ cell differentiation antigens (GDA) which appear at various stages of spermatogenesis and/or spermiogenesis. The GDA were found to be differentially expressed on spermatocytes, spermatids and on the various sub-regions of human spermatozoe. Some of the MoAbs also recognised changes in the antigenic structure of the sperm during their post-testicular maturation. Using solubilised sperm products, MoAb GDA-J/F1 reacted with a 92 kDa band in Western immunoblotting. The importance of these antibodies in the study of germ cell differentiation together with their potential clinical uses in the investigation of infertility, autoimmunity and immunological contraception are discussed.

Antibodies, Monoclonal↗

Immunoglobulins and monoclonal anti-blastocyst antibodies in the mouse uterine secretion during the early pre-implantation period.

A Sepharose bead blot technique was used to study immunoglobulins in the uterine secretion of mice during the pre-implantation stage. Secretion collected by Sepharose beads contained IgA, IgG, and IgM. The method could be made ten-fold more sensitive by using anti-mouse IgG or IgM conjugated to Sepharose beads. It has also been demonstrated that when injected intravenously, biotinylated purified immunoglobulins, both non-specific mouse myeloma IgG and IgM and specific anti-blastocyst IgG and IgM, is able to pass into the uterine cavity of the mouse. It was further shown that when injected systemically, anti-blastocyst antibodies can reach the blastocyst. Functionally active specific antibodies against morulae and/or blastocysts may, therefore, be able to influence the pre- and peri-implantation development of the embryo and could serve as a useful model for experiments directed towards the identification of immunological contraceptive procedures.

Animals↗

Vaccines against fertility.

The first evidence for the efficacy of a birth control vaccine in humans is now available from the Phase II trials on the human chorionic gonadotrophin vaccine in India. Several sperm antigens have been identified as potential contraceptive immunogens and zona pellucida antigens have been reported that reversibly control fertility.

Chorionic Gonadotropin↗

Fertility control in wildlife: humans as a model.

In a time of accelerated extinction rates and biodiversity loss, it may seem illogical to be concerned with methods limiting the reproduction of wild populations; however, there is an urgent need to inhibit the proliferation of a wide variety of species. The range of animals for which fertility control is desired makes the development of a single method impossible. The various reproductive strategies used by individual species, the desired outcome of contraceptive programs (reversible or irreversible; male or female directed) and our ignorance of the reproductive biology of many endangered species necessitate thorough species-specific investigations. As fertility control in humans is a reality and research into methods of fertility control is more advanced, humans serve as a model for developing contraceptive approaches for wild species. Population control by traditional methods (indirect or direct intervention by culling, poisoning, translocation, etc.) is increasingly unacceptable to the public, making human studies even more valuable for finding solutions to overabundant wild populations. This review compares and contrasts the range of contraceptive methods used in both wildlife and humans.

Animals↗

Recent developments in female contraception: LHRH.

During the last few years new approaches to female contraception based on LHRH and its analogs have been developed. The physiological significance of pulsatile LHRH release and its stimulation of the pituitary has been elucidated by recent studies in rhesus monkey. Immunization against LHRH results in complete inhibition of reproductive function in animals and may find as a useful method of long-term fertility control in domestic animals. Clinical studies have utilized this knowledge to treat infertile hypogonadal women with chronic intermittent low-dose of LHRH. The superactive stimulatory LHRH analogs, used to treat infertility, paradoxically proved to have antifertility effects. They induce desensitization of the processes responsible for gonadotropic and gonadal hormone secretion, mediated by specific LHRH receptors in the pituitary and gonad. While contraceptive effectiveness of luteolytic approach remains to be proven, inhibition of ovulation by intranasal LHRH analog administration or continuous LHRH infusion by programmed minipumps seem to provide safe and effective contraception in women.

Amenorrhea↗

Temporal trends in antibody production in captive grey, harp and hooded seals to a single administration immunocontraceptive vaccine.

The temporal production of antibody to a single-administration immunocontraceptive vaccine, known to be immunocontraceptive in free-ranging female grey seals (Halichoerus grypus), was studied in captive grey seals, harp seals (Phoca groenlandica) and hooded seals (Cystophora cristata). The vaccine is based on liposome delivery of porcine zona pellucida antigens. When measured by antigen capture, the response of hooded and harp seals to the vaccine was similar to the response of grey seals. Determination of antibody production by ELISA with protein A, ELISA with rabbit anti-seal immunoglobulin sera and SDS-PAGE after affinity chromatography confirmed the similarity in response to the vaccine by grey and harp seals, but suggested lower titers in hooded seals. The vaccine produced titers in captive, juvenile grey and harp seals known to be immunocontraceptive in wild, adult grey seals.

Animals↗

Molecular medicine and population control.

As we uncover the physiological and molecular mechanisms behind human reproduction, we gain the potential to exert more control over our reproductive capabilities. In the past two decades, the prospects for 'infertile' women to bear children, or 'sterile' men to father them, have improved dramatically; recently, women have given birth in their sixties, well beyond their natural menopause. Thanks to developments in contraception, couples can now enjoy an active sex-life without a significant risk of pregnancy, and more accurately control the size and timing of their families. But these new freedoms have a price: they also have the potential to cause demographic distortions, medical harm to individuals and abuse of human rights.

Contraception↗

Recent advances in contraceptive vaccine development: a mini-review.

Contraceptive vaccines (CV) may provide viable and valuable alternatives to the presently available methods of contraception. The molecules that are being explored for CV development either target gamete production [luteinizing hormone-releasing hormone (LHRH)/GnRH, FSH], gamete function [sperm antigens and oocyte zona pellucida (ZP)], and gamete outcome (HCG). CV targeting gamete production have shown varied degrees of efficacy; however, they either affect sex steroids causing impotency and/or show only a partial rather than a complete effect in inhibiting gametogenesis. However, vaccines based on LHRH/GnRH are being developed by several pharmaceutical companies as substitutes for castration of domestic pets, farm and wild animals, and for therapeutic anticancer purposes such as in prostatic hypertrophy and carcinoma. These vaccines may also find applications in clinical situations that require the inhibition of increased secretions of sex steroids, such as in uterine fibroids, polycystic ovary syndrome, endometriosis and precocious puberty. CV targeting molecules involved in gamete function such as sperm antigens and ZP proteins are exciting choices. Sperm constitute the most promising and exciting target for CV. Several sperm-specific antigens have been delineated in several laboratories and are being actively explored for CV development. Studies are focused on delineating appropriate sperm-specific epitopes, and increasing the immunogenicity (specifically in the local genital tract) and efficacy on the vaccines. Anti-sperm antibody (ASA)-mediated immunoinfertility provides a naturally occurring model to indicate how a vaccine might work in humans. Vaccines based on ZP proteins are quite efficacious in producing contraceptive effects, but may induce oophoritis, affecting sex steroids. They are being successfully tested to control feral populations of dogs, deer, horses and elephants, and populations of several species of zoo animals. The current research for human applicability is focused on delineating infertility-related epitopes (B-cell epitopes) from oophoritis-inducing epitopes (T-cell epitopes). Vaccines targeting gamete outcome primarily focus on the HCG molecule. The HCG vaccine is the first vaccine to undergo Phase I and II clinical trials in humans. Both efficacy and lack of immunopathology have been reasonably well demonstrated for this vaccine. At the present time, studies are focused on increasing the immunogenicity and efficacy of the birth control vaccine, and examining its clinical applications in various HCG-producing cancers. The present article will focus on the current status of the anti-sperm, anti-ZP, anti-LHRH/GnRH and anti-HCG vaccines.

Animals↗

Development of male contraceptive vaccine--a perspective.

This paper reviews the recent advances that have occurred in the area of development of a male contraceptive vaccine. The vaccine candidates considered for review are hormone/hormone receptor-based proteins including luteinizing hormone-releasing hormone (LHRH)/LH, follicle stimulating hormone (FSH), as well as LH and FSH receptor proteins. The review also highlights the advances in our basic understanding of gonadotrophin action which have led to development of these vaccines. Focus is mainly on studies in the non-human primate which may be directly relevant to projected studies in the human. The data indicate that the vaccines are well tolerated by the primate (including the human based on limited data) and do not give rise to any known toxic symptoms or immediate health hazards. The response to the immunogen has been uniform and it may be possible to increase antibody titres as well as prolong the immune response by adding acceptable immune stimulators to the adjuvant cocktail and developing better immunization schedules or immunogen delivery systems. Contraceptive vaccines for the male are a feasible proposition and attention should now be focussed on evaluating carefully the bioefficacy of antibodies raised to recombinant ovine FSHbeta or FSH receptor protein fragments in both human and non-human primates. The advantage of the FSH/FSH receptor over the LHRH/LH-based vaccine lies in the fact that the former does not require an exogenous testosterone supplement to maintain accessory gland function, libido etc. The LHRH/LH-based vaccine results in azoospermia, while the FSH vaccine causes the production of low numbers of poor quality spermatozoa which are incapable of impregnating cycling females.

Amino Acid Sequence↗

Implantation, interception and contraception.

The factors involved in post-fertilization events leading to implantation in mammals are discussed with special reference to potential forms of interception. The stages of embryonic growth until implantation are considered initially. The growth and differentiation of the uterine endometrium is then described, followed by the events occurring during the apposition and invasion of the implanting embryo. Several potential approaches to new forms of interception are considered, and the advantages and disadvantages of each of them are evaluated. Among them, new vaccines against the zona pellucida, inactivation of the secretions of the blastocyst, hatching, the activity of pinopodes, and the endometrial proteins produced in the secretory phase seem to offer various and varied targets. Some existing methods of fertility regulation may act by affecting these stages of development, e.g. RU486 may interfere with pinopod function. Various physiological and embryonic consequences of interfering with these stages of pregnancy are discussed.

Animals↗