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PubMed1980

The occurrence of sperm antibodies in human reproduction. I. Comparative new and improved test methods for sperm antibody detection.

The four essential methods for sperm antibody detection are considered and evaluated. The micro-sperm-agglutination and micro-sperm-immobilization tests have been improved and optimized. Quantitative estimation of sperm antibodies was made by an immunoelectrophoresis technique after separation of antibodies from the sera and genital tract secretions by affinity chromatography. Antibodies were typed by line-rocket immunoelectrophoresis. The pathologic errors of the modified tests were found to be small and the biologic errors were tolerable. Satisfactory results were achieved for sperm antibodies belonging to IgA and IgM, but not for other immunoglobulins.

Antibodies↗
PubMed1994

Antifertility effects of porcine zona pellucida-3 immunization using permissible adjuvants in female bonnet monkeys (Macaca radiata): reversibility, effect on follicular development and hormonal profiles.

Female bonnet monkeys Macaca radiata (n = 8, four per group) were immunized with purified 55 kDa glycoprotein from porcine zona pellucida (ZP3) and ZP3 conjugated to the beta subunit of human chorionic gonadotrophin (beta hCG) using adjuvants permissible for human use (alum, muramyl dipeptide and sodium phthalyl derivative of lipopolysaccharide). The animals were monitored for anti-ZP3 antibody titres, biweekly progesterone concentrations, menstrual cyclicity and status of fertility. All the animals generated a good anti-ZP3 antibody response, continued to have ovulatory cycles, remained infertile in the presence of high anti-ZP3 antibody titres and showed no disturbance in cyclicity (except summer amenorrhoea). Examinations by laparoscope showed normal ovaries with developing follicles or corpora lutea on the surface. Fifty per cent of the animals conceived after a decline in antibody titres. Ovaries of animals that failed to regain fertility were examined for changes in morphology at times when anti-ZP3 antibody titres in the circulation were low and following a booster when titres were high. None of the ovaries showed any sign of inflammation or lymphocytic infiltration. Follicles at different stages of development were seen in all of the ovaries. No significant reduction in the number of follicles, except in one monkey (MRA 178), was observed. There was no increase in the numbers of atretic or degenerating follicles. The results showed that ZP3 immunization with permissible adjuvants could be used for immunocontraception without obvious ovarian changes.

Adjuvants, Immunologic↗
PubMed2003

Zona pellucida glycoproteins based immunocontraceptive vaccines: strategies for development and their applications.

The mammalian oocyte is surrounded by an extra-cellular matrix, the zona pellucida (ZP), composed of three major glycoproteins (ZP1, ZP2 and ZP3). The ZP glycoproteins, by virtue of their tissue specificity and critical role during mammalian fertilization, have emerged as potential candidate antigens for the development of an immunocontraceptive vaccine. Molecular characterization of ZP glycoproteins from several species, reveals a variable degree of homology among the deduced primary amino acid sequences, which provided an opportunity to undertake active immunization studies in heterologous animal models. Active immunization of various animal species with either native ZP glycoproteins or those obtained by recombinant DNA technology led to the inhibition of fertility. Thus ZP glycoproteins based immunocontraceptive vaccines offer an attractive proposition for controlling wild life population. To make it a practical proposition, additional research inputs are required to optimize and devise novel strategies for vaccine delivery. Observed ovarian dysfunction, often associated with immunization by ZP glycoproteins is one of the major stumbling blocks for their use in humans. Ongoing studies to delineate appropriate B cell epitopes of ZP glycoproteins that are devoid of oophoritogenic T-cell epitopes, which will inhibit fertility without concomitant oophoritis, will be critical to determine their feasibility for human use.

Animals↗
PubMed1986

Formulation of a potential antipregnancy vaccine based on the beta-subunit of human chorionic gonadotropin (beta-hCG). IV. The role of adjuvants in booster injections.

Experiments were conducted in rabbits to determine the effect of adjuvant use on the antibody response following booster injections. The antigen used was in all cases the beta-subunit of human chorionic gonadotropin linked to tetanus toxoid (beta-hCG-TT). Adjuvants used were Al(OH)3, MDP analogs, and a streptococcus preparation, OK432. Primary vaccinations included Al(OH)3 adjuvant with or without supplementary adjuvants. In general, the greater the antibody response following primary vaccination, the greater the response following booster vaccination whether or not adjuvant was used in the booster. No increment in antibody titers was found by reason of including MDP analogs in booster vaccinations. OK432, in contrast, gave increased responses in booster injections which were in several cases statistically significant. The value of including Al(OH)3 in booster injections is not clear from the experimental data. In no case was the increment due to its inclusion large.

Adjuvants, Immunologic↗
PubMed2001

Absence of corpus luteum rescue by chorionic gonadotropin in women immunized with a contraceptive vaccine.

OBJECTIVE: To determine whether antibodies induced by a betahCG vaccine in women are competent to neutralize the luteotropic action of hCG. DESIGN: Prospective clinical study. SETTING: Hospitals and a laboratory at an academic center. PATIENT(S): Six immunized and three control women of reproductive age, participating in a clinical testing of betahCG contraceptive vaccine. INTERVENTION(S): Increasing doses of hCG simulating early pregnancy. MAIN OUTCOME MEASURE(S): Anti-hCG antibody titers, serum progesterone and urinary hCG levels, and onset of menses. RESULT(S): Administration of hCG to mimic early pregnancy sustained serum progesterone concentrations and extended the luteal phase in control women. In contrast, serum progesterone levels declined and the luteal phase was not extended if prevailing antibody titers were > or =40 ng/mL in women who had been immunized with a betahCG based vaccine. No booster effect was seen in anti-hCG titers after hCG challenge. CONCLUSION(S): Antibodies elicited by a betahCG vaccine inactivate hCG and prevent the hormone from rescuing corpus luteum, resulting in progesterone fall and normal menses. Lack of booster in the antibody response confirms the reversibility of the approach.

Adult↗
PubMed1987

A potential anti-pregnancy vaccine built by conjugation of the beta-subunit of human chorionic gonadotropin to adjuvant-active muramyl peptide.

The beta-subunit of human chorionic gonadotropin (hCG) conjugated to tetanus toxoid is being investigated as a vaccine for human fertility control. Initial clinical trials indicated that the level of antibody response induced by such an immunogen was not always sufficient to prevent pregnancy. Therefore, efforts are being made to evaluate new carriers for the beta-subunit and to select adjuvants to yield a more efficient vaccine. In the present report, we demonstrate that conjugates of the beta-subunit of hCG with muramyl dipeptide (MDP), or its nonpyrogenic derivative murabutide, may have potential as an effective antipregnancy vaccine. The copolymer of beta hCG and MDP administered with Al(OH)3 to mice induced a high anti-beta hCG response, better than that induced by the conjugate of beta hCG to tetanus toxoid given with Al(OH)3. Moreover, the antibodies induced by such an immunogen were competent for neutralizing the biological activity of hCG in vivo. Even more interesting, a copolymer of beta hCG and of murabutide induced high levels of biologically active antibodies. This immunogen may represent a promising candidate for the development of an efficient vaccine for human fertility control.

Acetylmuramyl-Alanyl-Isoglutamine↗
PubMed2005

Contraceptive vaccines targeting sperm.

Overpopulation is a global problem of significant magnitude, with grave implications for the future. Development of new contraceptives is necessary, as existing forms of birth control are unavailable, impractical and/or too expensive for many individuals due to sociological, financial or educational limitations. Immunocontraception and, in particular, the targeting of antibodies to sperm-specific antigens implicated in sperm-egg binding and fertilisation offers an attractive approach to control fertility. Sperm-specific antibodies may impair fertility by inhibiting sperm motility, by reducing penetration of the cervical mucus by sperm, or by interfering in sperm capacitation or the acrosome reaction; alternatively, antisperm antibodies may invoke the complement cascade, resulting in sperm lysis. The antibodies raised against sperm-specific antigens have proved to be extremely effective at reducing sperm-egg interactions in vitro; fertility trials in subhuman primates will eventually be needed to prove the effectiveness of the sperm antigens in terms of contraceptive efficacy before trials in humans can be justified. In addition, existing and emerging strategies (such as sperm proteomics, the determination of molecular and structural details of sperm proteins, and the modelling of protein-ligand interactions using X-ray and/or NMR structures to name a few) are expected to provide the experimental foundation for the design of small molecule inhibitors with antifertility effects. The technology underpinning vaccine development is constantly being developed and the introduction of DNA/RNA vaccines is certain to impact upon the field of immunocontraception.

Animals↗
PubMed2004

Effects of porcine zona pellucida immunocontraceptives in zoo felids.

Methods of contraception are necessary for management of zoo felids; however, the most commonly used contraceptive (melengestrol acetate implant) is associated with serious adverse reactions with long-term use. Porcine zona pellucida (pZP) vaccines are promising as contraceptives, but their safety in zoo felids has not been tested. pZP vaccine was administered to 27 female felids representing 10 species, including African lion (Panthera leo), Asian leopard (P. pardus), jaguar (P. onca), tiger (P. tigris), snow leopard (P. uncia), cougar (Felis concolor), Siberian lynx (F. lynx), Canada lynx (F. canadensis), serval (F. serval), and bobcat (F. rufus), in 15 facilities. Over 6 wk, each animal received three i.m. injections of 65 microg pZP with Freund's complete adjuvant (FCA), Freund's incomplete adjuvant, or carbopol as the adjuvant. Behavioral signs of estrus were seen in 14 of the vaccinated felids. An unacceptably high incidence of adverse reactions was seen including injection site swelling, lameness, limb swelling, or abscessation (or all) in five felids after injection with FCA as the initial adjuvant. Adverse behavioral signs, including increased irritability and aggression, were seen in four felids. Six of the felids were assayed for antibodies against pZP during the 12 mo after vaccination; all showed antibody production. Antibody levels appeared to peak 1-4 mo after vaccination began, although elevated antibody levels persisted in two animals for > 12 mo after the first injection. All vaccinated felids were ovariohysterectomized 3-13 mo after vaccination. Folliculogenesis was present in all treated animals, and there was no histopathologic evidence of inflammatory damage to ovaries. Contraceptive efficacy was not specifically evaluated in this study; however, two of the three felids housed with an intact male became pregnant during the study, one of which gave birth to healthy cubs.

Animals↗
PubMed1988

Immunological cross-reactivity of antibodies with species chorionic gonadotropin is a critical requirement for efficacy testing of human gonadotropin vaccines in sub-human primates.

A key question in the evaluation of a contraceptive vaccine is its efficacy in the prevention of pregnancy. Primates have been employed for evaluation of the efficacy of candidate vaccines against human chorionic gonadotropin (hCG). Discrepancies have been noted between the immuno-reactive antibody titres (against hCG) determined by RIA and their ability to prevent pregnancy in test animals. To gain further information on factors involved in fertility control, other characteristics of antibodies, such as affinity for hCG, bioneutralization capacities for hCG, hLH and monkey chorionic gonadotropin, have been determined in antisera from ten bonnet monkeys (Macaca radiata) at times when they were infertile and in bleeds preceding conception. The data reported show that the most important correlate for efficacy is the bioneutralization capacity of the antibodies for the species CG; animals became pregnant when the neutralization capacity diminished to low levels (less than 45 IU/l = 5 ng/ml of hCG equivalent). The bioneutralization of heterospecies CG decreased with the increase in affinity for hCG. These findings have implications for the choice and suitability of primate species for the various candidate vaccines being developed. Vaccines inducing highly specific antibodies against hCG, and restricted in their reactivity with primate CG, may not be testable in heterospecies primate models.

Animals↗
PubMed2004

Passive immunization for immunocontraception: lessons learned from infectious diseases.

Development of vaccine for contraception is an exciting proposition that could provide a valuable alternative to the presently available methods for birth control. Various targets such as gonadotropin releasing hormone (GnRH), follicle stimulating hormone (FSH), leutinizing hormone (LH), zona pellucida (ZP) antigens, sperm antigens, and human chorionic gonadotropin (hCG) are being explored for immunocontraception. Besides specific concerns associated with each contraceptive vaccine, the progress has been restricted by the variability of the immune response after active immunization, attain and maintain high antibody titers, time lag to achieve reasonably good antibody titers, and uncertainty regarding how long the bioeffective antibodies will remain in circulation. It is envisaged that these concerns may be taken care of by using the preformed antibodies in the passive immunization approach. The antibody therapies have been tried and found to be successful against various infectious diseases both in animals as well as humans. Some have become treatment modalities in the clinics. This manuscript will review the data available for the passive immunization of preformed polyclonal and murine/humanized/human monoclonal antibodies, their efficacy, mode of delivery, duration of the effects, and limitations, if any. The overall objective is to examine the feasibility and practicability of the passive immunization approach for immunocontraception.

Animals↗
PubMed2003

Molecular targets for immunocontraception.

Effective contraception is necessary in countries where limiting population growth has become a public policy imperative. The main antigenic targets for contraceptive vaccine development can be listed as following: 1) sperm antigens, 2) zona pellucida antigens, 3) gonadotrophin-releasing hormone, 4) chorionic gonadotrophin, 5) other protein/peptide hormones (follicle-stimulating hormone, luteinizing hormone, luteinizing hormone-releasing hormone), and 6) gonadal steroid hormones. New techniques, such as the application of monoclonal antibodies, hybridoma and DNA recombinant technologies, have become useful in search for contraceptive candidates. Current state of development of contraceptive vaccines based on specific antigenic targets and trials in animal/human models are presented in this article.

Animals↗
PubMed2003

[Specific mucosal immunity in the female reproductive tract: protection against infectious diseases and regulation of fertility (part 2)].

Immune mediated defense against pathogenic microorganisms and maintenance of immune tolerance to allogeneic antigens of sperm and fetus are main function of female reproductive tract mucosal immune system. Specific mechanisms concerning development of mucosal vaccines to sexually transmitted diseases and the immune regulation of fertility are reviewed in this article.

Animals↗
PubMed1976

Immunological block to mammalian fertilization: survival and organ distribution of immunoglobulin which inhibits fertilization in vivo.

Fertilization of golden hamster eggs was blocked both in vitro and in vivo by antibodies produced in rabbits against specific hamster ovarian antigens (HOA). Antibodies against HOA bound to surfaces of the hamster egg zona pellucida and prevented spermatozoa from attaching to the zona and entering eggs in vitro. Fertilization in animals could be blocked for four estrous cycles by a single intraperitoneal injection of anti-HOA immunoglobulin, but not by control immunoglobulin. The in vivo fate of anti-HOA immunoglobulin was analyzed by simultaneous injection of 125I-anti-HOA IgG and control 131I-IgG. Both anti-HOA IgG and control IgG appeared in a variety of organs (lung, kidney, spleen, liver, and uterus) shortly after injection, but disappeared rapidly with no detectable differences in organ half lives. However, in the ovary anti-HOA IgG (but not control IgG) persisted at high levels during the period of infertility. Quantitative precipitin analysis of organ homogenates indicated that a high percentage of anti-HOA IgG in the ovary (but not in the other organs tested) was immunologically indistinguishable from IgG indicating lack of radiolabel metabolism and reincorporation. The results are discussed in terms of the development of a specific immunological block to fertility.

Animals↗
PubMed2006

Effect of sperm DNA vaccine on fertility of female mice.

Our laboratory has identified a sperm-specific dodecamer peptide sequence, designated as YLP(12), vaccination with which causes a long-term reversible immunocontraceptive effect in female mice. In the present study, the effects of YLP(12) DNA vaccine were examined. YLP(12) 36 bp cDNA was cloned into pVAX1 vector to prepare the DNA vaccine. Two additional vaccine constructs were made by in frame cloning of one and two CpG repeats in the YLP(12)-cDNA vaccine. Five groups of female mice were immunized intradermally by using gene gun with YLP(12)-cDNA, YLP(12)-cDNA-CpG, YLP(12)-cDNA-CpG-CpG, YLP(12)-cDNA mixed with exogenous synthetic CpG oligodeoxynucleotide (ODN), or vector DNA alone, respectively. Vaccination with all three constructs and the YLP(12) vaccine mixed with exogenous ODN raised antibody response both in the sera as well as locally in the vaginal tract. There was no antibody response in the mice injected with the vector alone. In sera, the highest titers were obtained for the IgG class for all constructs and formulation followed by IgA class. In vaginal washings the highest titers were obtained for the IgA class followed by IgG class. Within the IgG class, the titers for the IgG2a subclass were significantly greater than the IgG1 subclass. Immunization with all constructs and formulation caused a significant (P < 0.05 to <0.001) reduction (20-43%) in fertility of female mice. The highest reductions were seen in mice immunized with YLP(12)-cDNA-CpG-CpG (two repeats) (43% reduction) and with the YLP(12) vaccine administered with exogenous CpG ODN (42% reduction). T lymphocytes obtained from DNA-vaccinated mice showed clearly distinguished comparative RT-PCR analysis of cytokine mRNA expression for Th1 and Th2 immune responses compared to T lymphocytes obtained from control animals injected with vector DNA. Expression of both Th1 cytokines (IL-2 and IFN-gamma) and Th2 cytokines (IL-4 and IL-10) was enhanced after DNA vaccination as compared to controls, with a bias towards Th1 response. The immunocontraceptive effects were long-lasting observed up to 1.3 years of the observation period and increased with time. These novel findings indicate that the intradermal immunization with a sperm-specific DNA vaccine causes a long-term circulating and local immune response resulting in immunocontraceptive effects in female mice.

Animals↗
PubMed2004

How far from a hormone-based contraceptive vaccine?

Antibodies of appropriate specificity are able to block the action of hormones which are obligatory for successful reproduction. Thus, if immunisation using such hormones can provoke adequate titres of bioneutralizing antibodies in sexually mature individuals, the vaccinee becomes infertile ('immunocontraception') for as long as sufficient titres of the antibodies are maintained. In the case of hormones that are required for the development of sexual maturity in the male, immunisation of young animals can prevent sexual maturation ('immunocastration'). The hormones which have been targeted are gonadotropin-releasing hormone (GnRH) for both immunocastration and immunocontraception, and follicle-stimulating hormone (FSH) and human chorionic gonadotropin (hCG) for immunocontraception.

Chorionic Gonadotropin↗