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Antisperm Contraceptive Vaccines: Where we are and where we are going?

As the developed and most of the developing nations have an infrastructure for mass immunization, the development of vaccines for contraception is an exciting proposition. The aim of this article isto review the current status and future prospective of CV targeting spermatozoa. CV that are being investigated target: Vaccines targeting gamete production affect steroid production.

They can be used as a substitute for castration of animals feral, farm, and domestic and for therapeutic purposes in clinical situations requiring inhibition of sex steroid secretion. Vaccines inhibiting gamete function are the preferred target. Although CV targeting ZP have a high contraceptive efficacy, they cause oophoritis, affecting sex steroids. For human applicability, the current research is focused on delineating infertility-related epitopes B-cell epitopes from oophoritis-inducing epitopes T-cell epitopes and modulation of immunogenicity by various carriers and adjuvants.

Presently, the focus is on increasing its immunogenicity and efficacy, and investigating its utility in HCG-producing cancer. Spermatozoa have drawn much attention for CV development. Spermatozoon can produce antisperm antibodies ASA in both men and women due to its auto- and isoantigenic properties.

ASA affect fertilization and fertility through several mechanisms, including: These women developed antisperm antibodies and no conception was reported for up to 1 year of observation. These findings provide strong evidence that spermatozoa are capable of eliciting an immune response that can cause a contraceptive state.

A US patent was issued for this spermatoxic vaccine in US patent number However, the whole spermatozoon cannot be used for the CV development as it has several antigens that are likely to be shared with various somatic cells. The utility of a sperm antigen for the CV development is contingent upon its: A sperm antigen which has these properties and is also involved in human immunoinfertility is a promising and exciting candidate for CV.

Immunoinfertile patients with ASA have no other tissue pathology concomitant with the infertility, indicating probable sperm-specificity of the molecule against which these antibodies are directed to.

The sperm-ZP binding site is the most desirable target for antisperm immunocontraception. The mouse genome is 2. It is about 14 percent shorter than the human genome, which is 2. The human genome is filled with more repeat sequences than the mouse genome. Almost every month there is a report on a new gene knockout that has some effect on fertility.

Using an extensive database search, we identified at least genes in mice whose knockout affect male fertility. Others did not demonstrate a surface expression, which is a must to make it accessible and amenable for antibody binding. Notable among them are: Although a sperm protein involving in any step of sperm function and fertilization is a viable candidate, the antigen involved in sperm-zona binding site is the most attractive molecule for CV development.

Our laboratory extensively investigated, using various technologies, the sperm proteins that are involved in zona pellucida ZP binding in humans. Human sperm proteins belonging to four major molecular regions, namely 95, 63, 51, and 14—18 kDa, were found to react with human zona pellucida proteins in the Western blot and immunoprecipitation procedures. Three 95, 51, and 14—18 kDa of the four molecular regions of sperm proteins that bound to the zona pellucida proteins also seem to involve o-phospho-L-tyrosine residues in their interaction.

These proteins demonstrated the presence of phosphotyrosine residues, and the kDa protein also showed autophosphorylating activity in the vitro kinase assay. It was found that the 51 kDA protein is the FA-1 antigen that was delineated in our laboratory.

In a recent study, the yeast two-hybrid Y2H system was used to identify human sperm proteins that interact with human ZP3. 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. Although there are over published studies in the database examining the effect of sperm antibodies on sperm function and fertilization in vitro , there are only three studies examining the effect of antibodies in vivo.

It is pertinent to demonstrate the effect of an antibody on fertility in vivo. There are only limited studies that investigated the contraceptive effect of well-defined sperm antigens in actively immunized animals.

The sperm antigens which have been examined for contraceptive effect include: Finding a suitable animal model to examine the effect of a vaccine on fertility, has been a major challenge for the reproductive immunologists working in the field of immunocontraception.

Most of the active immunization studies have been carried out in the mouse model. A few of the studies have used rat, rabbit, and guinea pig as a model. The rabbit and guinea pig are ultra-sensitive models to examine the immunogenicity and antifertility effect, and may not be suitable models for a vaccine to be tested in humans.

The mouse model is more acceptable, but it still has several concerns. The female mouse ovulates several over 20 eggs every cycle and a woman ovulates typically one egg every cycle. Therefore, there are differences between the mouse and human. This may also be because of an inherent nature of the mouse model, where it is challenging to make mice completely infertile.

However, after active immunization, one does find some mice that are totally infertile. Using the same antigen, one can observe a different effect on fertility in different strains of vaccinated mice. Mouse sperm do not bind to or penetrate human oocytes and vice versa. Two studies have examined the effect of a sperm antigen vaccination in a non-human primate model.

One study reported reduced fertility of female baboons after immunization with LDH-C 4. In another study, male monkeys were immunized with Eppin. The potential immunopathologic effects of immunization were not investigated. These findings indicate that CV can work for both males and females. Sperm antigens have auto-as well as isoantigenic potentials. For antisperm CV for males, ideally the vaccine should not affect spermatogenesis and sperm maturation or cause orchitis and epididymitis.

The blood-testes barrier should remain intact. The circulating antibodies percolate into the male genital tract via rete-testis, vas deferens, seminal vesicle, and prostate. Except for the few studies described above, most of the antisperm CV have been tested in female animals. One of the major challenges for development of an effective antisperm CV is to induce high titer antibodies locally in the genital tract, besides inducing high circulating titers in the serum.

For an effective immunocontraceptive, both circulating and local antibodies are required. These antibodies have to be present throughout the various parts of the genital tract. The female genital tract can also synthesize antibodies locally at several sites. However, our knowledge on the localization of various immune cells and components in various parts of the female genital tract are limited.

Not much research has been done in this area. Also, only a paucity of information is available on penetration of antibodies from circulation to different parts of the genital tract. In order to understand the immune correlates of contraceptive protection and to develop effective vaccination strategies, an understanding of the mucosal immunology of the genital tract is vital.

The female genital tract has immunological uniqueness and is different from other mucosal tracts. It has various components of mucosal immunity in different combinations from vagina to ovarian follicle, as examined in humans and animal models.

Cervical secretions are rich in IgA, IgG and secretory component. Endocervix has lamina propria with plasma cells that predominantly secrete IgA. Uterus is devoid of organized lymphoid tissues, but plasma cells secreting IgA and IgG are present. Oviducts have few, if any, plasma cells in the submucosal tissues, although IgA and IgG have been detected in the secretions. It has been shown that the majority of immunoglobulins present in the genital tract of healthy fertile women enter via transudation from the circulation.

To increase the immunogenicity of CV, especially the immune response in the genital tract, various routes of delivery, adjuvants, carriers including multiple carriers, and virus-like particles, have been tried. The recent focus is primarily on two aspects: DNA vaccines have been proposed as an exciting mode for vaccination and have several distinct advantages including easy manipulation, use of a generic technology, simplicity of manufacture, and chemical and biological stability.

The cytokines produced after a Th1 immune response, could inhibit gamete and embryo function since they express receptors for these cytokines. The effect was long-lasting up to at least one year of the observation period.

These are the first studies where contraceptive effect of sperm-specific cDNA vaccines were examined. To increase the efficacy of the vaccine, the constructs containing multiple repeats of the cDNAs, multiple repeats of in-frame CpG, and combined immunization of DNA and peptide vaccines are being investigated. The present trend is to use synthetic peptides for CV development instead of whole recombinant molecules. Although synthetic peptides require conjugation with T cell epitopes for immunogenicity, they are easy to synthesize in large amounts, are stable and easy to manipulate.

Also, these vaccines are helpful in enhancing immunogenicity. There is variability of immune response among individuals after any vaccination. Incorporation of multiple sperm epitopes in combination vaccines and use of different carriers and adjuvants can obliterate this concern.

Another approach to outstrip the variability is to use passive immunization employing preformed antibodies. Several of these antibodies have become treatment modalities in the clinics. The mouse monoclonal antibody can elicit strong anti-mouse antibody reaction; chimeric antibody can cause anti-chimeric response, and xenogenic complementarity-determining regions CDRs of humanized antibodies can also evoke an anti-idiotypic response, when injected into humans.

Phage display technology has been widely used to obtain a variety of engineered antibodies, including single chain variable fragments scFv antibodies against several antigens.

ScFv is an antibody fragment that plays a major role in the antigen-binding activity, and is composed of variable heavy VH and variable light VL chains connected by a peptide linker. The most widely used linker is Gly 4 Ser 3. The affinity and stability of the scFv antibodies produced in bacteria, are comparable with those of the native antibodies and are maintained by a strong disulfide bond.

ScFv antibodies can be produced on a large scale using specially modified bacterial hosts and have an advantage over the whole immunoglobulin Ig molecule. ScFv antibodies lack the Fc portion that eliminates unwanted secondary effects associated with Fc and, because of its small size, they can be easily absorbed into the tissues as well as genetically manipulated. Using the phage display technology and peripheral blood leukocytes PBL from ASA-positive immunoinfertile and vasectomized men, we have synthesized in vitro , for the first time, several human scFv antibodies that react specifically with well-defined sperm antigens that are involved in human sperm function.

Their contraceptive effect in vivo is being investigated. At this time, no immunocontraceptive is available on the market. These antibodies may provide a novel once-a-month, the first of its kind, immunocontraceptive s for human use.

In conclusion, development of antisperm contraceptive vaccines is an exciting proposition and may provide a viable alternative to other modalities of contraception. During the last two decades, significant progress has been made in the field of antisperm immunocontraception.

Several sperm antigens have been delineated, cloned, and sequenced that have function in sperm physiology and fertilization. Besides examining their role in immunocontraception, we have also gained knowledge regarding their development in the testes, gene regulation, and local and systemic immunogenicity pertinent to vaccine development.

The utility of a sperm antigen in CV development is contingent upon: The search for delineating such an antigen is extensively being performed in several laboratories. Finding such an antigen will be a major breakthrough and will move the field forward. In contrast to other vaccines, such as against infections and cancer, a contraceptive vaccine will be used by healthy individuals.

For a CV to be acceptable, it needs to be highly tissue-specific without any side effect and should provide close to full protection against pregnancy. It requires high bioefficacy and high tissue specificity. Also, the CV development is a multi-disciplinary approach requiring expertise in several fields including vaccinology, immunology, molecular biology, cell biology, and reproductive endocrinology. Andrology in the Nineties. Modern ART in the 's.

Consensus workshop on advanced diagnostic andrology techniques. Hum Reprod ; Guidelines on the application of CASA technology in the analysis of spermatozoa.

European Society for Human Reproduction and Embryology. J Androl ; What is normal semen quality? On the use and abuse of reference limits for the interpretation of semen analysis results. Hum Fertil Camb ; Acceptable variability in external quality assessment programmes for basic semen analysis. World Health Organization; Results of the American Association of Bioanalysts national proficiency testing programme in andrology.

Validation of sperm counting methods using limits of agreement. Particle distribution in low-volume capillary-loaded chambers. J Androl a; Capillary-loaded particle fluid dynamics: J Androl b; Statistical methods for assessing agreement between two methods of clinical measurement.

Passing H, Bablok W. A new biometrical procedure for testing the equality of measurements from two different analytical methods. Application of linear regression procedures for method comparison studies in clinical chemistry, Part I. J Clin Chem Clin Biochem ; The effect of the breeding season, cryopreservation and physiological extender on selected sperm and semen parameters of four ferret species: Reprod Fertil Dev ; Improved semen collection method for wild felids: Influence of image sampling frequency on the perceived movement characteristics of progressively motile human spermatozoa.

Fractal analysis of capacitating human spermatozoa. Minimum sperm trajectory length for reliable determination of the fractal dimension. The development of smoothing-independent kinematic measures of capacitating human sperm movement. Hum Reprod a; Effect of image sampling frequency on established and smoothing-independent kinematic values of capacitating human spermatozoa. Hum Reprod b; Laboratory investigation of the infertile male. Taylor and Francis Medical Books; Repeatability and variance analysis on multiple computer-assisted IVOS sperm morphology readings.

ESHRE special interest group for andrology basic semen analysis course: Biotechnic Histochem ; Morphometric dimensions of the human sperm head depend on the staining method used. Computer-aided sperm analysis CASA of sperm motility and hyperactivation. Methods in Molecular Biology. Springer Humana Press ; Maree L, van der Horst G.

Quantification and identification of sperm subpopulations using computer-aided sperm analysis and species-specific cut-off values for swimming speed. Biotech Histochem ; Definition of the optimal criteria for identifying hyperactivated human spermatozoa at 25 Hz using in-vitro fertilization as a functional end-point. Quantitative observations of flagellar motility of capacitating human spermatozoa. Characterization of hyperactivated motility by human spermatozoa during capacitation: Arch Androl ; Effect of seminal plasma on capacitation and hyperactivation in human spermatozoa.

The Fractal Geometry of Nature. WH Freeman and Company; Fractals and the analysis of growth paths. Bull Math Biol ; Mortimer ST, Mortimer D. Kinematics of human spermatozoa incubated under capacitating conditions. Hyperactivation of stallion sperm is required for successful in vitro fertilization of equine oocytes. Biol Reprod ; This article has been cited by 1 Quantitative assessment of heavy metal effects on sperm function using computer-aided sperm analysis and cytotoxicity assays Farren Hardneck,Gadieja Israel,Edmund Pool,Liana Maree Andrologia.

Cloete Animal Reproduction Science. Historical and futuristic developments in bovine semen technology P. Wu British Poultry Science. Bishop Reproduction, Fertility and Development. Kirkman-Brown Reproduction, Fertility and Development.

Soler Reproduction, Fertility and Development. Basic science of reproductive medicine. An indicator for intracytoplasmic sperm injection? How to cite this article: The future of computer-aided sperm analysis. Asian J Androl ;

Anti-sperm antibodies in can markedly reduce the likelihood of natural conception. GM-CSF and PDGF-BB) followed by increase in chemokines (e.g. IL-8, with various etiologies, including autoimmune, iso-immune, and cellular .. to two healthy children, have regular sexual intercourse and menstrual. Sperm separation methods that yield a higher number of motile spermatozoa are glass wool filtration or density gradient .. In order to give an overview, only the four most common .. tories AB, Gothenburg, Sweden) or ISolate. ® Braquet P, Touqui L, Shen TY, Vargaftig BB: Perspectives in plate-. Here we report our experience and a review of the literature on sperm sex chromosome aneuploidies in these two conditions. Head morphology analysis generated four size and four shape parameters, which Jaffe, N.; Toth , B.B.; Hoar, R.E. .. and iso-butylmethylxanthine- to the medium restored sperm capacitation.