Treatment of infertility caused by antisperm antibodies.

Twenty infertile couples with antisperm antibodies in the male or in the female partner were scheduled for treatment. In 15 couples the male and in 5 couples the female partner was the antisperm antibody carrier. In all the couples the result of the in vivo and in vitro sperm-penetration test was negative or poor. The SCMC-test was strongly positive in each of the couples. In all the couples IgG and IgA antisperm antibodies could be demonstrated on the spermatozoa or in the cervical mucus. It was postulated that antisperm IgA and not antisperm IgG is responsible for the penetration reduction of spermatozoa in cervical mucus and for the "shaking phenomenon" in the SCMC-test. Intrauterine inseminations, performed in 20 couples, resulted in four pregnancies. Condom therapy in three couples, for at lest 6 months, had no result. Two men were treated with 96 mg methylprednisolone per day for 7 days; this resulted in a slight decrease of the sperm-agglutination titre, but no pregnancy occurred.     

Inhibition of sperm penetration through human zona pellucida by antisperm antibodies

An in vitro penetration test using human spermatozoa, sera, and eggs stored in a highly concentrated salt solution was designed for examination of the effect of antisperm antibodies on the process of fertilization. Spermatozoa from a healthy fertile donor incubated in modified Biggers, Whiiten and Whittingham (BWW) medium containing 7.5% antisperm-antibody-negative serum, could penetrate through the zonae pellucidae of the stored eggs, but not when the spermatozoa from the same donor had been incubated in modified BWW medium containing 7.5% antisperm-antibody-positive serum. After the antisperm-antibody-positive serum was absorbed with washed spermatozoa, the sperm penetration was not blocked. Therefore, antisperm antibodies appear to block human sperm penetration through the human zona pellucida.     

Evaluation of serum antisperm antibodies in infertility

Aims and Objective

To evaluate the role of serum antisperm antibody (ASA) in infertility.

Method and Material

This study was conducted in the Department of Obstetrics and Gynecology, Pt. J.N.M. Medical College, Raipur (C.G.), India, from December 2006 to July 2008 over 105 selected couples with primary and secondary infertility attending the infertility clinic. Their detailed clinical history was taken. Physical examination and routine as well as special investigations like pelvic USG, follicular study, and hysterosalpingography were done in the female. Complete physical examination and semen analysis of male partners were done. Couples were subjected to post coital test (PCT) 2–6 hours after intercourse to rule out cervical factor. Serum ASA titer in both partners was detected by ELISA. Results were interpreted for qualitative evaluation. ASA-positive cases were treated with low-dose daily oral prednisolone for 3 months and evaluated in terms of ASA titer, semen analysis, PCT result, and conception rate. The results were analyzed by statistical methods.

Results

Out of 105 couples, serum ASA-positive males were 38 (39.19%), of which definite serum ASA positive were 9 (8.57%), borderline (equivocal) were 29 (27.61%), and negative were 67 (63.08%). Among females serum ASA positive were 42 (40%), in which definite ASA positive were 19 (18.09%), borderline 23 (21.9%), and negative 63 (60%). Asthenospermia was found more common in ASA-positive men (55.56%, p=0.0001). Poor PCT was most commonly associated in husband ASA negative and wife ASA positive. Treatment with low-dose oral prednisolone resulted in significant increase in motility of sperms in male partners and decrease in ASA titer in both the patients. Pregnancy was achieved in 45.23% ASA-positive females, while among couples with ASA-positive husbands, 31.57% of wives conceived.

Conclusion

Serum ASA are considered to be cause of unexplained infertility and unexplained abnormal PCT. Antibodies against sperm prevent their motility through female reproductive tract and hamper the process of fertilization. Low-dose prednisolone was useful in infertility associated with ASA by improving sperm quality and giving rise to pregnancies.     

Characterization of human sperm antigens and antisperm antibodies in infertile patients.

OBJECTIVE: To identify which sperm antigens may elicit the production of functionally important antisperm antibodies. DESIGN: Immunoblot analysis was performed on 69 serum and 9 seminal plasma samples from infertile patients, using detergent extracts of pooled donor sperm as the antigen source. Serum and seminal plasma had been previously tested by an indirect immunobead binding test (IBT); 61 IBT-positive and 17 IBT-negative samples were included in the study. Proteins recognized by IBT-positive but not IBT-negative samples were most likely to be cell surface antigens, whereas proteins recognized by both IBT-positive and IBT-negative samples were probably intracellular. Antibodies directed toward surface antigens would be most likely to affect fertilization. Characterization of sperm surface proteins on both acrosome-intact and -reacted sperm used labeling of cell surface proteins with an N-hydroxysuccinimide ester of biotin, fractionation of sperm heads and tails, and lectin binding to determine glycosylation. RESULTS: Specific immunoreactivity (with respect to IBT results) was observed to 35K, 40 to 45K, 57K, 66K, and 88 to 90K MW proteins. Characterization studies identified an 88K MW glycosylated plasma membrane protein, a 66K MW inner acrosomal membrane protein, a 34K MW inner acrosomal membrane protein, and a 35K MW prominent tail protein. CONCLUSION: Immunological infertility may involve several antigens characterized in this study. Further studies are necessary to determine if antibodies to these specific proteins interfere with sperm function.     

Methods for the detection of antisperm antibodies associated with immunologically-mediated human infertility

Among others, immunological factors can be the reason for human infertility. The aim of the present critical review was to the analyze the literature data, published in the last 20 years, as well as our experimental and clinical results concerning the most frequently used in the international centers methods for the detection of antisperm antibodies associated with immunologically--mediated human infertility.     

Antizona and antisperm antibodies in women with endometriosis and/or infertility

OBJECTIVE: To measure the levels of antigamete antibodies in serum and peritoneal fluid of women with endometriosis and/or infertility. DESIGN: Antibody activity against human sperm and porcine oocytes was analyzed in selected subgroups of women. SETTING: Clinic of reproduction. PATIENT(S): Women with endometriosis and/or infertility. INTERVENTION(S): No treatment was implemented before peritoneal fluid and blood sample collection. MAIN OUTCOME MEASURE(S): Quantitative ELISA. RESULT(S): Four groups of women (n = 98) were analyzed for the presence of antizona and antisperm antibodies: infertile with endometriosis (n = 30), idiopathic infertility (n = 28), fertile with endometriosis (n = 20), and healthy fertile controls (n = 20). Antibodies were analyzed simultaneously in serum and peritoneal fluid. No statistically significant differences in antibody levels were detected in serum samples among the analyzed groups. The median values for antizona and antisperm antibodies in peritoneal fluid were significantly higher in women with idiopathic infertility than in the control group. In women with unexplained infertility, a high degree of correlation (Spearman) was found between the presence of antizona antibodies in peritoneal fluid and serum (r = 0.579). A positive predictive value of 80% was calculated for the presence of antizona antibodies (>5 ng/oocyte) in the peritoneal fluid of patients with infertility. CONCLUSION(S): Antizona antibodies locally produced in the peritoneal fluid have diagnostic value for infertility status; however, they cannot be treated as a marker or prognostic factor for minimal endometriosis and/or its treatment.     

Identification of human sperm antigens reacting with antisperm antibodies from sera and genital tract secretions.

OBJECTIVE: To identify sperm antigens reacting with antisperm antibodies relevant in human infertility. DESIGN: The reactions of separated sperm antigens with antibodies present in sera and genital tract secretions from infertile and fertile females and males were examined by immunoblotting techniques. SETTING: The patients were followed in an outpatient setting of a hospital clinic. PATIENTS: One hundred consecutive infertile males and females, referred for determinations of antisperm antibodies, comprised the study group. Fifty hospital and faculty employees with proven fertility served as a control group. RESULTS: A high proportion of sera from fertile and infertile humans contained antibodies reacting with at least one sperm antigen. However, two discrete bands of antigenic proteins with molecular weights of 44 and 72 kd reacted significantly more frequently with serum antibodies from infertile females than from fertile females. No apparent correlation could be demonstrated between any particular antigen and serum antibodies from infertile males. Nevertheless, antigenic proteins of 62 kd were identified as the major sperm antigens reacting with antibodies present in seminal plasmas from infertile males. CONCLUSIONS: The major sperm antigens reacting with systemic antibodies differ from the antigens recognized by local antisperm antibodies. Sperm antigens exhibiting relative molecular weights of 62 kd are major antigens reactive with local antisperm antibodies from infertile humans.     

Inhibition of human sperm-zona pellucida and sperm-oolemma binding by antisperm antibodies

The results of this preliminary investigation suggest that antisperm antibodies interfere predominantly with sperm-zona pellucida binding. The observation of similar numbers of control (Ab-) and test (Ab+) sperm bound to the oolemma implies that the antibody-mediated inhibition of capacitation, acrosome reaction, or oolemma binding may not be major causes of failed fertilization with sperm autoimmunity. However, only seven patients were studied, and further investigation with larger numbers of subjects are required.     

Therapeutic approaches to the treatment of infertility associated with antisperm antibodies production

A lot of extensive investigations have been conducted to evaluate the potential role of antisperm antibodies (ASA) in infertile couples. But many questions still remain on the role of ASA in reproduction, testing methods, significant levels of ASA and treatment of immunologically-mediated human infertility, due to ASA. The aim of the present critical review was to the analyze the literature data, published in the last 20 years, concerning the most frequently used in the international centers methods for treatment of human immunological infertility associated with ASA production.     

抗精子免疫性不孕的研究进展

<正>不明原因不孕症是一种临床难治性疾患,不少研究提示免疫功能异常是不明原因不孕症的重要致病因素。免疫性不孕是指不育夫妇有抗生育免疫证据存在,并排除其他不孕致病因素。自1959年Rumke发现不孕症患者血清中存在抗精子抗体(AsAb)以来,AsAb与女性不孕症的相关性已引起人们的关注。     

Capacitated sperm cells react with different types of antisperm antibodies than fresh ejaculated sperm.

OBJECTIVE: To determine if sera of some women have antibodies against capacitated but not freshly ejaculated sperm. DESIGN: The sera of 66 women undergoing in vitro fertilization (IVF) were tested for sperm antibodies after 1 hour and 18 hours of sperm incubation in the maternal sera. Subsequently, 5 sera were tested with capacitated versus noncapacitated sperm cells. SETTING: The study was carried out in a university hospital department. PATIENTS, PARTICIPANTS: The patients were 66 consecutive couples undergoing IVF. INTERVENTIONS: Sera and semen that were taken for routine tests as part of the IVF procedures were used. MAIN OUTCOME MEASURES: A case with IVF failure associated with late appearance of sperm antibodies prompted us to study the detection of sperm antibodies after 1 hour and 18 hours incubation. RESULTS: Of 37 cases negative for sperm antibodies after 1 hour incubation, 7 demonstrated high levels of antibodies after 18 hours incubation. In 21 of 23 cases with low or intermediate levels of antibodies after 1 hour incubation, significantly higher levels (P less than 0.05) of antibodies were found after 18 hours. Different and higher levels of sperm antibodies were observed in five sera after incubation of 1 hour with capacitated sperm as compared with noncapacitated controls. CONCLUSIONS: Major antigenic differences may exist between capacitated and noncapacitated sperm. In some women sperm antibodies are reactive against capacitated sperm only. This has no certain clinical significance but may explain certain cases of IVF failure, unexplained infertility, and part of the variation in sperm antibodies testing methods.     

Intrauterine insemination as a treatment of infertility in women with antisperm antibodies.

Twenty-four women with infertility caused by antisperm antibodies were treated by homologous intrauterine insemination. Initially, all the women had timed intrauterine insemination by washed spermatozoa for three cycles. The pregnancy rate per couple was 4.20%. The remaining 23 patients received a combined treatment of chlomiphene citrate and intrauterine insemination for three cycles, which did not increase the pregnancy rate per couple and per cycle (4.3% and 1.4% respectively). Thereafter, the remaining 22 patients received a combined treatment of hMG and intrauterine insemination for another three cycles which resulted in a pregnancy rate per cycle (6.1%) and per couple (18.20%) that was significantly greater (P less than 0.01). We conclude that infertile women with antisperm antibodies can benefit after a trial of induction of multiple follicular development with hMG in combination with intrauterine insemination.     

Characterisation of a sperm coating auto-antigen reacting with antisperm antibodies of infertile males using monoclonal antibodies

Objective In a previous study a number of sperm-specific antigens were identified which reacted with antisperm antibodies from both infertile and vasovasostomised males. To investigate the localisation and distribution of these antigens and their role in male fertility, monoclonal antibodies were raised against them; immunoblotting techniques were used to select only those antibodies which competed with human antisperm antibodies for these human auto-antigens.
Design One antibody, NW21, reacted with an 18 kDa auto-antigen present on epididymal sperm but absent from testicular sperm. Immunohistochemical studies showed that the antigen is produced in small basal cells between the columnar epithelium of the corpus epididymis, passes up into the tubule and then coats sperm passing along the epididymis. Sperm stored in the cauda epididymis and ductus deferens stain strongly for this sperm coating glycoprotein.
Conclusions The localisation of this antigen supports the suggestion that auto-immune infertility may represent a response to epididymal rather than testicular sperm. Monoclonal antibodies raised to unique and immunologically accessible sperm coating proteins, produced in the epididymis rather than in the testis, would seem to present an excellent theoretical solution to male contraception.     

Frequency of anti-sperm antibodies demonstrated by classical tests for sperm antibodies in patients with unexplained infertility

The aim of the present investigation was to establish the frequency of sperm antibodies in patients with etiologically unexplained infertility, and to compare the demonstrated frequencies with the results from y control group of y healthy blood donors, as well as with the results of other investigators. The gelatin agglutination test of Kibrick and the tray agglutination test of Friberg were applied to test 244 sera from infertile patients and 50 sera from healthy blood donors at the Laboratory of Immunology of Reproduction, Department of Biology, Medical University of Sofia. For the infertile patients, relevant sperm antibody titers were demonstrated in 2.5% (titer > or = 16) for the Kibrick method, and in 7% (titer > or = 32) for the Fiberg method. The test of Kibrick did not reveal significant antibody titers in the healthy controls, while the test of Friberg showed sperm antibodies in 2% of the blood donors. Fisher's exact test demonstrated extremely significant correlation (p < 0.0001) between the presence of sperm antibodies in sers of patients with unexplained infertility as revealed by the tests of Kibrick and Friberg. Most often mixed agglutinates were demonstrated in the Friberg test. In contrast with the results of other investigators head-to-head agglutinins were observed more often in male sera, while tail-to-tail agglutinins--in female sera. Finally, the results from the present investigation, as well as the analyzed literature data showed a low frequency of anti-sperm immunity in the Bulgarian population. The established high degree of correlation between the tests of Kibrick and Friberg, the good reproducibility of the results and the low cost of these methods confirm their appropriate use for the diagnosis of sperm antibodies in patients with unexplained infertility.     

Detection of antisperm antibodies: their localization to human sperm antigens that are transferred to the surface of zona-free hamster oocytes during the sperm penetration assay

The authors have developed an extension of the sperm penetration assay for detecting serum immunoglobulins to sperm antigens that are transferred to the plasma membrane of a sperm-penetrated hamster oocyte. After the hamster oocytes have been scored for sperm penetration by observing for the presence of swollen sperm heads, they are incubated in serum followed by either a 20-minute treatment with rhodamine-conjugated protein A (which binds to most subclasses of IgA, IgG, and IgM) or a 2-hour incubation in guinea pig serum (complement). Positive fluorescence indicates that the serum contains antibodies to sperm antigens that were transferred to the surface of an oocyte during gamete fusion. Complement-mediated lysis indicates that the immunoglobulin that is bound can also fix complement. The advantages of this assay for detection of serum antisperm antibodies are that it is an extension of a widely used assay, is rapid and requires readily available reagents and equipment, can detect most subclasses of IgA, IgG, and IgM, detects antibodies to those sperm antigens that may be transferred to the oocyte during fertilization, and indicates whether the detected antisperm antibodies can mediate complement-dependent lysis of the fertilized oocyte.     

Seminal fluid antisperm antibodies do not reflect those present on the sperm surface

Given the increasing evidence that head-directed antibodies can impair fertilization in vitro, as well as play a role in the impaired entry of sperm into cervical mucus, our findings provide strong support for the direct analysis of immunoglobulins bound to the sperm surface, rather than by indirect analysis through the study of seminal fluid.     

Macrophages and infertility: oviductal macrophages as potential mediators of infertility

Human peritoneal macrophages have previously been shown to phagocytize normal sperm. We had hypothesized that if macrophages were present in the distal oviducts, they could interfere with fertilization by phagocytizing sperm in vivo. The present study was designed to determine whether functional macrophages are present in the human oviducts, and to determine the relationship between oviductal and peritoneal macrophages. Forty patients undergoing laparotomy for sterilization or evaluation of infertility or other gynecologic factors were studied. Infertile patients with endometriosis had more peritoneal macrophages than did fertile normal women or infertile women with distal or proximal tubal obstruction. Oviductal macrophages were observed in all patients. The oviductal macrophages were indistinguishable from the peritoneal macrophages, as judged by similar morphologic features, adherence to plastic, phagocytosis of polystyrene spheres and IgG-coated erythrocytes, and presence of peroxidase and alpha-naphthylbutyrate esterase. Patients with endometriosis had the highest numbers of oviductal macrophages, while those patients with distal tubal obstruction had extremely few oviductal macrophages. The results suggest that oviductal macrophages may arise from peritoneal macrophages that migrate into the oviducts.