Antisperm Autoantibodies Research Articles

The Possible Role of SARS-CoV-2 in Male Fertility: A Narrative Review

The spread of severe acute respiratory syndrome—Coronavirus 2 (SARS-CoV-2) around the world has rapidly sparked the interest of the scientific community to discover its implications in human health. Many studies have suggested that SARS-CoV-2 is directly or indirectly involved in the male reproductive tract impairment. Some evidence supports the possible role of the virus in male infertility. Therefore, this review aims to summarize the relationship between the male urogenital tract, male fertility, and the gonadal hormone profile. The testis is one of the organs with the highest expression of the angiotensin-converting enzyme (ACE) 2-receptor that allows the virus to penetrate human cells. Orchitis is a possible clinical manifestation of COVID-19 and testicular damage has been found on autopsy in the testes of patients who died from the disease. SARS-CoV-2 infection can compromise the blood-testis barrier, favoring testicular damage and the production of anti-sperm autoantibodies. Some studies have detected the presence of SARS-CoV-2 in semen and a high percentage of patients with COVID-19 have altered sperm parameters compared to controls. Finally, lower testosterone levels, higher luteinizing hormone (LH) levels, and decreased follicle-stimulating (FSH)/LH and testosterone/LH ratios suggest primary testicular damage. In conclusion, further studies are needed to evaluate the exact mechanisms by which SARS-CoV-2 affects the male reproductive system and fertility and to evaluate the reversibility of its long-term effects.

Overweight and obesity: The allies of prostate inflammation.

To the Editor, World Health Organization (WHO) defines infertility as the inability of a sexually active partner to achieve a clinically confirmed pregnancy after at least one year of intercourse without contraceptive protection on fertile days. As women are primarily responsible for the conception, childbirth, and postpartum, fertility problems have been related to fertile female capacity. However, men play an essential role, being responsible for about 50% of alterations in fertility due to dysfunctions in the male reproductive tract, including varicocele, hypogonadism, poorly descended testicles, testicular tumors, and even anti-sperm autoantibodies [...].

Pathogenesis of Autoimmune Male Infertility: Juxtacrine, Paracrine, and Endocrine Dysregulation.

According to global data, there is a male reproductive potential decrease. Pathogenesis of male infertility is often associated with autoimmunity towards sperm antigens essential for fertilization. Antisperm autoantibodies (ASAs) have immobilizing and cytotoxic properties, impairing spermatogenesis, causing sperm agglutination, altering spermatozoa motility and acrosomal reaction, and thus preventing ovum fertilization. Infertility diagnosis requires a mandatory check for the ASAs. The concept of the blood–testis barrier is currently re-formulated, with an emphasis on informational paracrine and juxtacrine effects, rather than simple anatomical separation. The etiology of male infertility includes both autoimmune and non-autoimmune diseases but equally develops through autoimmune links of pathogenesis. Varicocele commonly leads to infertility due to testicular ischemic damage, venous stasis, local hyperthermia, and hypoandrogenism. However, varicocelectomy can alter the blood–testis barrier, facilitating ASAs production as well. There are contradictory data on the role of ASAs in the pathogenesis of varicocele-related infertility. Infection and inflammation both promote ASAs production due to “danger concept” mechanisms and because of antigen mimicry. Systemic pro-autoimmune influences like hyperprolactinemia, hypoandrogenism, and hypothyroidism also facilitate ASAs production. The diagnostic value of various ASAs has not yet been clearly attributed, and their cut-levels have not been determined in sera nor in ejaculate. The assessment of the autoimmunity role in the pathogenesis of male infertility is ambiguous, so the purpose of this review is to show the effects of ASAs on the pathogenesis of male infertility.

Fluoride Induces Autoimmune Orchitis Involved with Enhanced IL-17A Secretion in Mice Testis

Fluoride (F) widely exists in the water and food. Recent studies reported that F induced testicular toxicity via inflammation reaction. This study was aimed to explore the mechanism of F-induced inflammation in testis. 100 healthy male mice (BALB/cJ strain) were randomly divided into five groups including: control, experimental autoimmune orchitis (EAO), and three F groups (25, 50, and 100 mg/L sodium fluoride (NaF)). After 150 d, the results showed a significant increase in testicular cytokines levels including of IL-17A, IL-6, IFN-γ, and TNF-α in NaF and EAO groups compared with control group. Interestingly, the presence of specific antisperm autoantibodies in antitesticular autoantibodies and the notable recruitment of immunocyte (T cells and dendritic cells) were also observed in NaF and EAO groups. In addition, findings showed that in NaF and EAO groups macrophages and T cells both significantly secreted IL-17A, and the protein and mRNA levels of cytokines (IL-6 and TGF-β) were significantly increased. From these results, it can be concluded that autoimmune orchitis and IL-17A are implicated in F-induced testicular inflammation.

Celiac Disease Occurrence with Autoimmune Infertility in Infertile Men

Objectives: The aim of the present study was to determine the prevalence of celiac disease (CD) among unexplained infertile men and determine the association of CD autoantibodies with antisperm autoantibodies (ASA) as diagnostic tool for Immunological Infertility. Materials and Methods: One hundred and sixty six male patients were suffering from primary unexplained infertility were chosen to diagnose CD. Three autoantibodies (AD) were detected by performing enzyme linked immunosorbent assay (ELISA), deamidated gliadin peptide/tissue transglutaminase (DGP/tTg-IgA), tTg-IgG, and IgA. Other two autoantibodies were detected by ASA, IgG and ASA-IgA to diagnose immunologic infertility. All serologic procedures and seminal fluid examinations were done on unexplained infertile men in Basrah Governorate, South of Iraq. Results: Out of 166 participants with unexplained infertility, 15 participants (9%) have been diagnosed as immunological infertile. Thirteen participants (7.8%) have silent CD. Only seven (4.2%) have both immunologic infertility and CD. Statistically significant association between CD and immunological infertility (OR)>20 with (95%CI) between 7.2-58. Significant correlation (p<0.05) was resulted between the five autoantibodies and grade A+B motility parameter of seminal fluid analysis. Conclusion: Screening for CD should be part of the diagnostic work-up of unexplained infertile men, and immunological infertility should be considered as additional AD associated with CD in infertile men.

Antisperm Antibodies in Undescended Testis

Background: Undescended testis is the commonest genital malformation in boys. Men with a history of undescended testis have a reduced probability of fertility.Aim of the study: To explore whether the development of anti-sperm antibodies and antitesticular autoantibodies might play a role in inhibiting the future fertility of cryptorchid boys. Histopathological and hormonal assessments were done for those patients.Patients and methods: The study group consisted of 30 patients with undescended testis who underwent Sub dartous orchidopexy at The AL-Kindi Teaching Hospital and other private hospitals. Orchidopexy and histopathological studies were done for them. Hormonal assessment, antisperm and antitesticular autoantibodies assessment were done for them.Results: The mean age of the orchiopexy patients was 12 ± 9.4 years. Histopathological study revealed testicular atrophy in 76.6% of the cases. Antitesticular autoantibodies were detected by direct immunofluorescence. Antisperm antibodies and hormonal assay in patients’ serum before and after operation illustrated no significant difference.Conclusions: The late descent of a testicle into the scrotum may impair its development ending in reduced fertility because of histopathological changes (atrophy) already become apparent in the first few months of life. Development of antitesticular autoantibodies may contribute in this process because the ectopic position of testis that damage barrier. Therefore, hormonal and surgical treatments complement each other and should be provided before the child’s first birthday.

A Conservative Approach to Testicular Rupture in Adolescent Boys

Management for blunt trauma with breach of the renal capsule or bladder (extraperitoneal) has largely become nonsurgical since a conservative approach proved to be effective and safe. Currently the recommendation for managing testicular rupture is surgical exploration and débridement or orchiectomy. We report outcomes in boys diagnosed with testicular rupture and treated without surgical intervention. In the last year we conservatively treated 7 consecutive boys with delayed presentation of testicular rupture after blunt scrotal trauma. Patients were treated with scrotal support, antibiotics to prevent abscess, rest, analgesics and serial ultrasound. We report clinical information and outcomes. The 7 boys were 11 to 14 years old and presented 1 to 5 days after injury. Trauma was to the left testis in 3 cases and to the right testis in 4. Patients presented with mild to moderate pain and similar scrotal swelling. Ultrasound findings consistently revealed hematocele and increased echogenicity. Blood flow was present in the injured portion of the testes in 3 cases and to the remainder of the affected testicle in 6 of the 7 boys. In the remaining boy an adequate waveform was not seen in either testicle, which the radiologist thought was secondary to prepubertal status. Other findings included scrotal edema, irregular contour and seminiferous tubule extrusion. Followup was greater than 6 months in all cases. Five boys were seen at the office and the 2 remaining had telephone followup. In all cases hematocele resolved, testicular size stabilized without atrophy and echogenicity normalized in the 5 patients with followup ultrasound. One patient required surgical repair of hydrocele 4 months after trauma but no other patient needed surgical exploration. No abscess or infection developed. A conservative approach in a select group of adolescent boys with testicular rupture can result in resolution of the fracture and maintenance of testicular architectural integrity.

The effect of the antisperm auto-antibody-bound sperm on in vitro fertilization outcome.

To evaluate the effects of antisperm auto-antibody-bound sperm on the outcome of in vitro fertilization-embryo transfer (IVF-ET), 160 infertile couples undergoing treatment by in vitro fertilization were recruited in this study. In the study group (11 couples, 15 cycles), the male partners were positive for antisperm autoantibodies determined by immunobead test (IBT). In the control group (149 couples, 152 cycles), the men had no such antibodies. The percentages of fertilization rate, cleavage rate and pregnancy rate of the study group and control group wer 75.0 +/- 5.2% vs. 69.3 +/- 2.4%; 82.8% +/- 3.7% and 6.7% +/- 11.8%, respectively. There were no significant differences in in vitro region, type and/or percentage of sperm-bound antibodies also had no effect on the in vitro fertilization outcome. In conclusion, in vitro fertilization-embryo transfer is not significantly affected by antisperm autoantibody-bound sperm determined by immunobead test.

Immunological Usefulness of Semen Manipulation for Artificial Insemination Homologous (AIH) in Subjects with Antisperm Antibodies Bound to Sperm Surface*/Immunologische Nützlichkeit der Spermaaufbereitung für eine homologe artifizielle msemination bei Män

The authors report trials carried out in the preparation of semen for homologous artificial insemination in subjects with antisperm autoantibodies. The tests have demonstrated that swim-up is not able to select spermatozoa free of antibodies bound to the sperm surface. All the experiments of migration, dilution, antigenic competition and cryoconservation, carried out to evaluate the theoretical possibility of a better recovery of non "immunologically compromised" spermatozoa, have confirmed such negative results both with the Immunobead Test and Cytofluorimetric analysis.

Distribution of Molecular Epitope for Ts4, an Anti-Sperm Auto-Monoclonal Antibody in the Fertilization Process

To investigate molecular effects of anti-sperm autoantibodies on fertilization, we previously established anti-mouse sperm-head auto-monoclonal antibodies (mAbs). Among the mAbs established, one mAb (named Ts4) recognized the sugar moiety of TEX101, a germ cell-marker glycoprotein. In the present study, we examined the immunoreactivity of Ts4 in mouse spermatozoa and fertilized eggs during early embryogenesis to clarify the distribution of the Ts4-reactive antigen in the fertilization process. Similar to TES101 mAb (a specific probe for TEX101), immunopositive staining of Ts4 was observed on spermatocytes, spermatids and spermatozoa within the testis. In contrast to the results obtained with TES101 mAb, Ts4 reacted with the sperm acrosomal region within the cauda epididymis. A Western blot analysis of epididymal sperm extract revealed that Ts4 mainly detected two bands between 100 and 150 kDa, while Ts4 faintly detected a band corresponding to TEX101 at 38 kDa. In addition, Ts4-reactive molecules were observed in the growing early embryo after fertilization. Since Ts4-reactive antigen, potentially a carbohydrate chain, is only observed in reproduction-related areas such as the testis, epididymal sperm-head and early embryo, it is expected to have an effect on fertilization. Therefore, additional studies of this antigen may elucidate the molecular mechanisms underlying the reproductive process.

Antisperm autoantibody response after unilateral vas deferens ligation in rats: when does it develop?

Unilateral obstruction or injury to the vas deferens can result in significant injury to the contralateral testicle. Antisperm autoantibodies are thought to play a significant role in this phenomenon. It has been reported that early surgical repair of the vas, before the development of antisperm autoantibodies, will prevent any potential damage to the contralateral testicle. This led us to investigate the timing of the antisperm antibody production and to attempt to determine whether antibody production precedes histologic testicular damage in the Lewis rat model. In a controlled study, mature rats were divided into temporal groups, with the experimental animals all receiving a unilateral vasectomy. At postoperative endpoints of 1, 7, 15, or 30 days, blood samples were collected for immunologic assay, and the testicles were harvested for histologic examination. Antibody levels were measured by an immunobead test using goat anti-rat immunoglobulin G (IgG)-coated Sepharose beads; tissue sections were fixed in Bouin solution, embedded in paraffin, and stained with hematoxylin and eosin. There was no statistically significant histologic difference between any of the groups. However, immunologic evaluation revealed a statistically significant increase in immunobead antibody binding in the 30-day group compared to the control groups (P = .02). These data seem to indicate that in this model, antisperm antibody production is not evident until 15-30 days after unilateral injury to the vas deferens occurs, and the development of these antibodies precedes any demonstrable histologic damage to the testicle. If it is correct to infer that human antisperm antibody production will also precede histologic testicular damage, and further, that the onset of the human autoantibody response may vary from several days to weeks, then in cases of suspected or known ductal injury, the clinical monitoring of antisperm antibody levels could enable testicular damage to be predicted prior to its development and thus be avoided.

Outer dense fiber proteins are dominant postobstruction autoantigens in adult Lewis rats.

Obstruction of the male reproductive tract commonly results in generation of antisperm autoantibodies. However, only a few of the sperm autoantigens recognized by these antibodies have been characterized. To identify postobstruction rat sperm autoantigens, sperm proteins were separated by two-dimensional(2-D) gel electrophoresis. Spots corresponding to proteins that were stained by at least 50% of postvasectomy rat sera on 2-D Western blots were removed from polyacrylamide gels and microsequenced by tandem mass spectrometry. From a total of 21 spots, 12 contained peptides that matched solely to either of two outer dense fiber proteins, odf1 or odf2. Six additional spots contained peptides comprising odf1 or odf2 and were accompanied by peptides representing other proteins. Only three spots lacked outer dense fiber peptides but did contain sequences of other known proteins. The results indicate that the outer dense fiber proteins odf1 and odf2 are dominant postobstruction autoantigens because they were detected in the majority of the immunoreactive protein spots examined. Possible explanations for this observation include the abundance of outer dense fiber proteins in spermatozoa, slow solubility, which may provide a sustained supply of antigen, and testis-specific expression during spermiogenesis.

Antisperm autoantibody response is reduced by early repair of a severed vas deferens in the juvenile rat

Objective: To determine whether antisperm autoantibody production after prepubertal vas injury is influenced by immediate repair of the vas compared to delay of the reanastomosis until sexual maturity. Design: Animal study comparing early repair, late repair, and sham-operated groups. Setting: Research laboratory in a medical school. Patient(s): Lewis rats. Intervention(s): After division of the vas deferens in juvenile rats, animals in an early repair group had the vasa repaired immediately by using an absorbable intraluminal stent. Animals in a late repair group had vasa obstructed by ligation until after puberty, when they underwent microsurgical vasovasostomy (age 60 days). Main Outcome Measure(s): Antisperm antibodies were assayed by ELISA. The weights of reproductive organs were determined, and samples of testis were studied by light microscopy. Result(s): The antisperm antibody response was less when the vas was repaired immediately than if the repair was delayed until after puberty. There was a low incidence of testicular alteration in the repair groups and none in sham-operated animals. Conclusion(s): If the vas deferens is injured or obstructed prepubertally, there may be a benefit to considering immediate repair to reduce the likelihood of developing antisperm autoantibodies, which have been associated with reduced fertility.

Post-obstruction rat sperm autoantigens identified by two-dimensional gel electrophoresis and western blotting

Although antisperm autoantibody responses to obstruction of the male reproductive system have been documented, information on the nature of the cognate sperm autoantigens has been limited. In the present study, the patterns of sperm autoantigens recognized by sera from rats after obstruction of the vas deferens or epididymis were studied by high resolution two-dimensional (2-D) gel electrophoresis and western blotting. Comparisons of patterns of autoantigens stained on 2-D western blots of sera from prepubertal vasectomy, prepubertal epididymal ligation and adult vasectomy groups revealed both similarities and differences. Sera from sham-operated animals showed no detectable reaction or much lighter staining of a small number of spots. Visualization of sperm autoantigens on 2-D western blots supported the hypothesis that there is a relatively small set of sperm proteins that can be regarded as dominant post-obstruction sperm autoantigens because they are recognized by multiple post-obstruction sera. The 2-D analysis revealed previously undetected distinctions in the autoantigens recognized after adult and prepubertal vasectomy, as well as variations with the site of obstruction. These differences in the response may be due in part to changes in antigens of spermatozoa in different parts of the tract and at different ages, as well as variations in exposure of sperm cell proteins to the immune system resulting from the sites of spermatic granulomas. Preparative 2-D gels and western blotting with post-obstruction sera are now being used to identify specific sperm autoantigens by microsequencing of selected proteins.

Fertilization antigen-1 removes antisperm autoantibodies from spermatozoa of infertile men and results in increased rates of acrosome reaction

Objective: To determine if fertilization antigen (FA)-1 will remove autoantibodies from the surface of sperm cells of immunoinfertile men by immune adsorption and permit an increased acrosome reaction (AR). Design: Prospective analytic study. Setting: University medical center. Patient(s): Men from 18 infertile couples with autoantibodies present on their spermatozoa. Intervention(s): Sperm samples after processing were examined for antibody binding and AR before and after adsorption with control medium or FA-1. Main Outcome Measure(s): Sperm-bound antibody was assessed by the immunobead assay (immunoglobulin [Ig] A and IgG) and the AR by induction with ionophore A23187. Result(s): Adsorption with FA-1 compared with control medium increased immunobead-free swimming sperm an average of 50% and 76% for IgA and IgG antisperm antibodies, respectively, with 78% and 100% of the 18 semen specimens increasing significantly. The AR rate increased an average of 10.3% compared with control medium and showed improvement in 78% of the sperm samples after FA-1 adsorption. Conclusion(s): The FA-1 sperm antigen appears to significantly free sperm cells coated with autoantibodies in the semen of most infertile men examined. Reducing sperm-bound antibodies that inhibited the AR allowed the sperm cells to undergo successful AR induction by calcium ionophore.

Degeneration of the seminiferous epithelium following epididymal obstruction in prepubertal rats.

The development of the testes was studied in rats following prepubertal obstruction of the epididymis. Male rats received bilateral ligation of the corpus epididymidis or a sham operation at 10 days of age, and temporal changes in testicular morphology and weights of reproductive organs were determined at intervals spanning sexual maturation. Development of the testes was normal through 35 days of age. The initial histological changes in the testes of ligated animals, observed at 56 days, included an increased diameter of the seminiferous tubule lumen, depletion of spermatids, and the presence of multinucleate spermatids. Subsequently, germ cells were greatly depleted in the testes of 91- and 128-day-old rats with ligated epididymides. After puberty, testicular weight and volume declined relative to corresponding sham-operated animals. On the other hand, the weights of the epididymides in ligated animals prior to puberty significantly exceeded those of sham-operated rats but weighed significantly less than those of rats in the sham group after sexual maturation. Testicular alterations occurred after increases in the weights of the epididymides. Testicular changes may have contributed to rather than resulted from an autoimmune response to spermatozoa because testicular alterations preceded increases in antisperm autoantibodies.

Epididymal obstruction during development results in antisperm autoantibodies at puberty in rats.

An autoimmune response to sperm occurs after vasectomy, but there is little information on whether similar reactions occur after obstruction of the male reproductive tract at other points. Male Lewis rats received bilateral ligation of the corpus epididymidis or a sham operation at age 10 days, and the subsequent systemic antisperm autoantibody responses were compared to those observed following obstruction of the vas deferens. After sexual maturation, rats with epididymal ligations had antisperm antibodies on an enzyme-linked immunosorbent assay that were significantly higher than those of sham-operated animals and did not differ from antibody levels in vasectomized rats at the same ages. Western blot analysis showed that certain sperm proteins were recognized by antisperm antibodies after both epididymal ligation and vasectomy, including the previously identified "dominant" autoantigens at 73-83, 68-72, 48, 42, and 22 kDa. On the other hand, sera from rats with epididymal ligations recognized 60 and 52 kDa proteins that were not bound by most postvasectomy sera. Conversely, 42-48 and 38-42 kDa bands were more strongly and frequently stained after vasectomy than after epididymal ligation. The results demonstrate that antisperm antibodies are produced after obstruction of the epididymis and that the magnitude of the response is comparable to that after vasal obstruction. Differences in autoantigens recognized after epididymal and vasal obstructions may reflect maturational changes in sperm components that take place during the passage of spermatozoa through the epididymis.

Antisperm antibodies in men with psychogenic anejaculation.

Antisperm autoantibodies were determined in 16 men suffering from psychogenic anejaculation who underwent assisted reproduction treatments. Blood and semen samples were collected after transrectal electroejaculation and antisperm antibodies in serum and on the surface of motile spermatozoa were measured using the direct and indirect immunobead binding test. Five men (31%) were found positive for antisperm antibodies. The majority of antibodies were directed against the sperm heads. Surface antibodies were mainly IgA isotype whereas serum antibodies were IgG isotype. These results suggest that psychogenic anejaculation might be associated with increased incidence of antisperm autoimmunity.

Semen analysis and antisperm antibody.

Evaluation of the fertilizing potential of sperm is an important procedure in in vitro fertilization and embryo transfer (IVF-ET) clinic, in order to avoid performing invalid conventional IVF-ET or unnecessary intracytoplasmic sperm injection (ICSI). However, none of the standard semen parameters are reliable indices in predicting IVF outcome. The test examining sperm-zona interaction is the best present method, though the short supply of the human zona component should be solved. IVF-ET treatment is useful for pregnancy in women having sperm immobilizing antibodies because it avoids exposing sperms to antibodies that block fertilization. ICSI should be used in immunologically infertile men diagnosed by specific method to demonstrate the in vivo exposure of sperms to antisperm autoantibodies that block fertilization.

Appearance of ‘natural’ antisperm autoantibodies after sexual maturation of normal Lewis rats

Serum antisperm antibodies were assessed quantitatively with an ELISA in normal male Lewis rats at intervals between ages 10 and 128 days, spanning the onset of puberty. Antisperm antibodies rose between 56 and 91 days, and were significantly higher in 91- and 128-day old rats than at earlier intervals. The animals underwent normal pubertal development as indicated by increases in weights of the seminal vesicles and ventral prostate. The rise in antisperm antibodies correlated temporally with events in the postnatal development of the male reproductive system, with the increase in antisperm antibodies most closely following the time when spermatozoa reach the epididymis and proximal vas deferens at approximately 56 days. The observation that serum antisperm antibodies increased only after sexual maturation suggests that some differentiation antigens of sperm are processed and presented to the immune system under normal circumstances in this strain. Western blot analysis showed that the sera from normal postpubertal Lewis rats bound several proteins, including bands of > 100, 75–82, 78, 68, 65, 63, 54–55, 42, 37, 35, 26, and 20–22 kDa. The majority of these autoantibodies were sperm-specific as shown by the absence of comigrating bands in western blots of somatic tissue extracts, although antibodies in postpubertal sera recognized certain other proteins in somatic tissues. Several protein autoantigens, defined by sera from postpubertal animals, matched dominant autoantigens recognized by antibodies produced in response to vasectomy, prepubertal vas obstruction, or immunization with spermatozoa. This finding indicates that the antisperm antibody responses following sperm immunization, vasectomy or prepubertal vasal obstruction represent accentuation of an autoantibody response to sperm that develops normally following puberty.