Effects of varicocelectomy on anti-sperm antibody in patients with varicocele

Anti-sperm antibody [ASA] can decrease sperm motility and, therefore, it is a cause of male infertility. The aim of this study was to evaluate the effects of varicocelectomy on anti-sperm antibody in patients with varicocele. This observational study was conducted on 90 patients with varicocele at Sina and Imam Khomeini hospitals during 2006 to 2009. All varicocelectomy candidates were selected for ASA assessment both in semen and serum before and after surgery. ASA level was measured using a direct method for semen and an indirect method of Sperm MAR test, for serum. Paired t-test and McNemar's test were used for data analysis, and p<0.05 was considered statistically significant. ASA level in semen was 13.7% before, and 15.7% after three month of varicocelectomy [p=0.881]. Serum level of ASA before and after surgery were 13.6% and 21.7%, respectively [p=0.033]. Three parameters including sperm count, motility and morphology showed recovery following, varicocelectomy, but only the difference in sperm motility was significant [p<0.05]. This study showed that varicocelectomy has no effect on semen ASA. Although serum antibody has been shown to increase after varicocelectomy but sperm motility will improve. Varicocelectomy seems to have a beneficial effect on semen parameters in infertile men with varicocele

Antisperm antibodies in women with unexplained infertility

Unexplained infertility occurs in many couples of childbearing age, immune mechanisms have been postulated in this disorder for the last few decades. Circulating antibodies against spermatozoa present in serum and seminal plasma have been especially implicated. This autoimmunity against spermatozoa has been investigated in males, while the isoimmunity [in the females] has got low attention. Fifty women with unexplained infertility and twenty fertile women were involved in this case-control study. ELISA test was used to detect antisperm antibody [ASA] from cervical mucus [CM] and serum specimens of both groups of women. CM was collected at mid-cycle period and dissolved mechanically [not by bromeline]. Thirty percent of infertile women have IgG-ASA in their serum and 20% have IgA-ASA in the CM, while 22% of fertile women have IgG-ASA in their serum and no fertile women have any titer of IgA-ASA in their CM specimens. Only CM-IgA-ASA of infertile women showed significant statistical correlation with cellular property of CM, which was scored according to Insler score. It is concluded that ELISA test is more sensitive and specific than microagglutination tests for detection of serum and secreted ASA. Also, secretory IgA-ASA are more indicative amid have potential role in immunological infertility as iso- immunity than IgG-ASA. Therefore, it is strongly recommended that immunological infertility should be considered as an important cause of infertility and have to gain a special interest by clinicians

Incidence of anti-zona pellucida and anti-sperm antibodies among infertile Jordanian women and its relation to mycoplasmas

Anti-zona-pellucida autoantibodies [AZP-Ab] and anti-sperm isoantibodies [ASA] were assessed in the cervical secretions from 73 infertile Jordanian women and 41 fertile control women using latex agglutination. Significantly more women with infertility had AZP-Ab and ASA [16.4% and 8.2% respectively] compared with fertile women [9.4% and 0%], with no relation to the etiology of infertility. Using polymerase chain reaction Mycoplasma hominis and Ureaplasma urealyticum were detected in cervical secretions of 19.2% and 13.7% of infertile women, and the presence of mycoplasma was significantly correlated with the presence of AZP-Ab and ASA

[Sperm exposure and development of preeclampsia]

It is proposed that the excessive immunological response of mother to feto-placental unit follows by preeclampsia. To investigate whether the sperm exposure can reduce the incidence of preeclampsia by increased tolerance of mother to semen antigens. This was a prospective cohort study carried out on Iranian primigravid women receiving prenatal care at health clinics of hospitals affiliated to Isfahan University of Medical Sciences in 2006. The study population was divided into two groups marked as sperm-exposed group [n= 143] and non-exposed group [n=250]. The data were collected using a questionnaire including information such as time interval between the beginning of sexual contact and conception, duration of sperm exposure, and the development of preeclampsia. Statistical analysis was performed using t-student test, chi square test, and logistic regression. Our results showed that the rate of preeclampsia in sperm-exposed group was significantly lower than that of non-exposed group [p=0.043]. An inverse relationship between the length of exposure and the occurrence of preeclampsia was demonstrated [p=0.03]. Exposure to sperm seems to offer protection against development of preeclampsia and preconception sexual contact over a long period could reduce the risk of preeclampsia

Influence of antisperm antibodies in the semen on intracytoplasmic sperm injection outcome

OBJECTIVE: The aim of this study was to analyze the influence of autoantibodies against spermatozoa present in the semen on the outcome of in vitro fertilization with intracytoplasmic sperm injection (ICSI). MATERIALS AND METHODS: We performed a retrospective analysis of clinical and laboratorial data from a six year-period ICSI cycles. Screening for the presence of ASA in the semen, by using the direct immunobeads test (IBT), was available for 351 cycles. According to the percentage of antibody-bound spermatozoa in the semen, we divided the cycles in four groups: I (n = 194): 0 percent-10 percent ASA; II (n = 107): 11 percent-20 percent; III (n = 33): 21 percent-50 percent and IV (n = 17): 51 percent-100 percent ASA. Additionally, a group of 349 ICSI cycles performed with ejaculated spermatozoa from oligo/asthenozoospermic men who had insufficient number of motile sperm available for ASA screening was included for comparison. ICSI outcomes were compared among groups and included fertilization rate (2 PN), cleavage rate, cleavage velocity, embryo quality, clinical pregnancy and miscarriage rates. Data were examined statistically, with an alpha level of 5 percent considered significant. RESULTS: Fertilization, cleavage rate and velocity, percentage of good quality embryos, as well as clinical pregnancy and miscarriage rates did not differ among different ASA levels groups. ICSI outcomes in men exhibiting different levels of autoimmunity against spermatozoa did not differ from those with severely abnormal seminal parameters. CONCLUSIONS: Our data indicate that intracytoplasmic sperm injection (ICSI) outcomes are not influenced by ASA levels on sperm.

[Designing an ELISA method using sperm antigens for detection of antisperm antibody in comparison with SpermMar test

The role of antisperm antibodies with a prevalence of 6-26% is well known in immunological infertility. Thus, there is clinical importance to determine ASA levels in both male and female. Nowadays, one of the most important discussed controversies in the field of immunological infertility is establishing an standard method to determine ASA. It seems that ELISA method will be more sensitive, specific and more diagnostic in determination of ASA if sperm surface antigens could be used as coated antigens, with least contamination to sperm intracellular antigens and nonspermic antigens. So, the aim of this study is designing an ELISA method by using the best method of sperm antigens extraction with at least contamination. In this study we designed an ELISA method with three different extraction methods of sperm antigens including sonication method, using SDS detergent, and application of LIS detergent, then we compared ELISA method based on the three extraction methods as well as two similar commercial ELISA kit [IBL Co, and Bioserv Co] with SpermMar test. Comparing designed method with commercial kit indicated that among 28 sera which had 16 positive sera and 12 negative sera by SpermMar, 14 sera were true positive by LIS method and only 2 cases were false negative without any false positive results, whereas there were 5 true positive results and 11 cases false negative by the sonication method. The SDS method also had 13 true positive results with 3 false negative and 4 false positive results. In addition, two commercial kit had in turn 7 and 4 cases true positive and both of them had 1 case false positive and in turn 9 and 12 cases with false negative result. ELISA method designed by LIS detergent has adequate sensitivity [87.5%] with higher specificity [100%] and efficacy [92.8%] than other extraction methods. There is a significant correlation between this designed method and SpermMar test [r=0.572]. The results of this study indicated that ELISA method by LIS antigens has at least contamination with nonspermic antigens and it is better than other extraction methods and commercial ELISA kits for detection of antisperm antibody

[Correlation between in vitro fertilization with the level of antisperm antibody in seminal plasma measured by flow cytometry]

Antisperm antibodies [ASA] are present in%8-21 of infertile men. In vitro Fertilization [IVF] has been recommended as an effective procedure in couples with immunological male factor. Although this procedure has been found to bypass the inhibitory effect of antisperm antibodies on fertilizing ability of spermatozoa but the fertilization rate is reduced about%40 for ASA positive samples. The goal of present study was to investigate the correlation between anti-sperm antibodies measured by indirect flow cytometry and fertilization rate in infertile couples undergoing in vitro Fertilization [IVF]. Semen samples were collected from 80 infertile men undergoing IVF cycle in Isfahan fertility and infertility center. Couples were classified based on fertilization rate into high and low groups. 52 couples had high [>50%] and 28 couples had low fertilization rate [

Immunochemical and functional characterization of a polyclonal antibody to human sperm antigen.

A polyclonal antibody was raised against a 16 kDa human sperm protein identified by a monoclonal antibody to human sperm. The antibody showed significant reactivity with mouse spermatozoa as seen by ELISA. Immunohistochemical analysis showed that the antibody reacted with antigens from mouse testis, prostate as well as seminal vesicle. In both mouse and human testis the antibody localized antigens in round as well as elongated spermatids and mature spermatozoa. By SDS-PAGE and Western blot analysis the antibody reacted with a 16 kDa protein in the testis and seminal vesicle, whereas in the prostate it identified two proteins, one at 20 kDa and another at 25 kDa. Immunofluorescent localization by the antibody showed reactivity with acrosomal and/equatorial and midpiece region of human spermatozoa. The antibody showed extensive agglutination both in mouse and human spermatozoa. The results indicate that the antigen may be a conserved antigen. Cross reactivity of the antibody with mouse spermatozoa enabled us to carry out antifertility trials. Passive immunization of female mice with this antibody caused 67% reduction in fertility. It is likely that the antifertility effect could be partly due to agglutinating nature of the antibody which may have caused inhibition of all processes that depend on forward motility such as cervical mucus penetration and possibly preventing sperm egg interaction. Such well characterized and functionally relevant antibodies will enable to identify sperm antigens relevant for fertility. Identification of such antigens may also help in diagnosis of immuno infertility.

Anticuerpos antiespermatozoides en parejas con infertilidad de causa no explicada / Antisperm antibodies in couples with infertility of unknown origin

Se estudió una muestra de 75 parejas infértiles en relación con la presencia de anticuerpos antiespermatozoides en plasma seminal, moco cervical o adheridos a los espermatozoides, mediante la prueba de inmunobeads y se determinó la capacidad inhibidora del plasma seminal sobre la proliferación linfocitaria, para evaluar la capacidad inmunomoduladora del semen como participante en el mecanismo patogénico primario de aparición de los anticuerpos antiespermatozoides. Se organizaron las parejas en 2 grupos, según la presencia o no de anticuerpos antiespermatozoides y se compararon en cuanto a la frecuencia de embarazo y la capacidad inmunosupresora del plasma seminal. Además se comparó la incidencia de embarazo en grupos de parejas con capacidad inmunosupresora y sin ella. Se observó que la presencia de anticuerpos antiespermatozoides fue menos frecuente que la reportada por otros autores y no estuvo relacionada con la capacidad inmunosupresora del plasma seminal, tampoco influyeron sobre la frecuencia de embarazo la presencia de anticuerpos antiespermatozoides y la capacidad inmunosupresora del plasma seminal(AU)

Does intrauterine insemination in Saudi female cause production of antisperm antibodies?

To determine the possibility of inducing antisperm antibodies in patients undergoing intrauterine insemination with motile sperms from the husband. Clinico-immunological prospective study. King Abdulaziz University Hospital. Fifty patients undergoing Intrauterine Insemination [IUI], were tested for developing antisperm antibodies using agglutination test, complement mediated immobilization test, and immunoglobin specific indirect immunobead assay. Incidence and type of antisperms antibodies. Forty seven out of 50 patients remained negative for antisperm antibodies after 2-6 cycles of IUI. Detection of antisperm antibodies after IUI was evident in 3 patients [6 percent]. Intrauterine insemination does not appear to cause a significant or lasting immune response

Anticuerpos de la clase IgG unidos a la superficie espermática: correlación metodológica, cuadro citológico / Antisperm antibody binding on the sperm surface: statistical correlation between method, cytologycal characteristics

Entre el 5 y 10 por ciento de los hombres infértiles presentan en sus eyaculados anticuerpos antiespermatozoides (AcAe) en valores que superan las tasas aceptadas, por lo que la detección de este tipo de anticuerpos debe realizarse al iniciar los estudios de una pareja infértil. En este trabajo se emplearon eyaculados procedentes de hombres integrantes de parejas infértiles, con el objetivo de a) establecer la correlación entre dos métodos utilizados en la detección y cuantificación de AcAe con capacidad de unión a la membrana espermática, los Immunobeads test (IB) y el Mixed Antiglobulin Test: MAR test (Mt) y b) determinar las características citológicas de los eyaculados de hombres con alta tasa de anticuerpos con capacidad de unión a la membrana del espermatozoide. Para el estudio comparativo de ambos test se emplearon 58 muestras. El criterio de selección empleado es que uno de los dos tests superara los valores de referencia. La evaluación citológica se realizó en 250 muestras tomadas al azar y en 53 eyaculados leucospérmicos (concentración de leucocitos polimorfonucleares) (LPN) > 10 a la sexta. Método: Peroxidasa. Los IB para la detección de anticuerpos de clases IgG (IBG) e IgA (IBA) y Mt fueron desarrollados según normas de la OMS, empleándose la técnica directa en todas aquellas muestras con concentración espermática mayor a 5.10 a la sexta espermatozoides/mL y movilidad traslativa (grados [a] + [b]) superior a 40 por ciento, en los restantes casos se usó el método indirecto. En los 58 sémenes seleccionados, los anticuerpos de tipo IgG presentaron valores de (x+/-DS) 88,9+/-15,8 y de 58,8+/-31,6 para Mt e IBG respectivamente (P < 0,001). En el 86,2 por ciento de los casos (n=50) ambos se correlacionaron dando resultados francamente positivos, en el 13,8 por ciento (n=8) frente a IB negativos, se obtuvieron Mt positivos, no habiéndose obtenido valores dudosos en las muestras en estudio. Los valores obtenidos en los dos métodos directos que involucran a la IgG, se correlacionaron significativamente (r=0,58). Los resultados obtenidos con IB en los 58 pacientes seleccionados, mostraron en el 63,8 por ciento (n=37) de los sémenes anticuerpos IgG e IgA, en el 22,4 por ciento (n=13) sólo de clase IgG, no encontrándose ninguna muestra con anticuerpos de clase IgA, únicamente...

Identification of bovine sperm specific polypeptides reactive with antisperm antibodies.

The present study was taken to characterize molecular weights of sperm specific polypeptides antigenic to rabbits and calf with the aim to assess their immunoreactivity with IgG antibodies in sera from immuno-infertile cows. Seropositivity for antisperm IgG antibodies in 75 repeat breeder and 15 pregnant control cattle was tested by cellular ELISA using washed spermatozoa antigen from 4 bulls. Molecular weights of bovine sperm polypeptides antigenic to rabbit and calf were determined by 10% SDS-PAGE and Western blotting. Molecular weights of sperm peptides reactive with sera from immuno-infertile cows were also determined. Seropositivity of antisperm IgG antibodies for bull I, II, III and IV was 23.6, 14.6, 26.6 and 20%, respectively. A total of 16 polypeptides were discernible on gel. Out of these, 7 polypeptides were immunoreactive with sera from hyperimmunized rabbits as compared to 3 poly-peptides which reacted with sera from hyper-immunized calf. Only two polypeptides were reactive with sera from immuno-infertile cows. Variable number of sperm polypeptides and their immunoreactivity have been reported in different species. Antigenicity of different polypeptides in sperm needs further investigations.

Immunological assessment of infertility by estimation of antisperm antibodies in infertile couples.

160 clinical samples were collected from 40 infertile couples with unexplained infertility. The samples collected included serum and seminal plasma of the male partners and serum and cervical mucus samples of the female partners. 25 fertile healthy couples were investigated as controls. All the samples collected were then tested for class-specific antisperm antibodies by an Enzyme linked immunosorbent assay (ELISA). Antisperm antibodies were detected in 30% of the infertile couples which included 25% female and 10% male partners. Amongst the cases positive for antisperm antibodies, antibodies were detected most frequently in female sera 58.4% followed by male sera 33% and 25% in cervical mucus. The isotyping of antisperm antibodies in various samples showed IgG to be the most frequent type specific antibody followed by IgM & IgA types of antibodies. ELISA has provided a relatively simple, reliable and highly reproducible method of detection of antisperm antibodies. Thus application of antisperm antibody testing especially in cervical mucus should become an integral part of the investigation of immunologic infertility.

Study on serum sperm agglutination in cases of unexplained infertility.

In our study we investigated 100 couples of unexplained infertility in order to detect the presence of anti-spermatozoal antibodies. Both auto-immunity and Iso-immunity have been found responsible in 12.5% of couples of primary infertility & 10% of couples of secondary infertility. As many as, 21% of cases of unexplained infertility were attributed to presence of antibodies in sera of infertile couples. Here, we highlight the importance of anti-spermatozoal antibodies testing in the first instance itself in cases of unexplained infertility.

Integrin cell adhesion molecules on human spermatozoa.

The expression of integrin cell adhesion molecules (ITG-CAMs) by human ejaculated spermatozoa (fresh, capacitated and acrosome reacted) was evaluated by immunocytochemical, immunofluorescence and cell-ELIS methods, using monoclonal antibodies against alpha-6 and beta-3 subunits. Both the subunits were expressed on the acrosome region in fresh spermatozoa and post acrosomal region after acrosome reaction induced by calcium ionophore. The spermatozoa of the fertile men showed significantly (P < 0.001) higher expression of alpha-6 and beta-3 ITG subunits than the subfertile men. The percentage of spermatozoa reacting with alpha-6 and beta-3 mAbs increased significantly after the loss of acrosome when compared with fresh spermatozoa. Moreover, 35-40% of spermatozoa with normal shape and none of the spermatozoa with pathological shape showed a positive reaction. The quantitative analysis carried out by ELISA suggests that the levels of these ITG subunits decreased significantly (P < 0.001) in the subfertile subjects when compared with the fertile and the difference was more for alpha-6 than the beta-3. Hence our result suggests that alpha-6 subunit may be used as a clinical marker to evaluate the sperm quality in men.

Assessment of antisperm antibody in sera of mercury and lead exposed workers

Occupational exposure to lead or mercury was found to make protein better antigens. The production of autoantibodies to nervous system protein is one example of such effect. The present study aims to detect the possibility of induction of antisperm autoantibodies due to occupational exposure to lead or mercury. Male workers exposed to lead [n=50] or to mercury [n=39] were selected for this purpose and compared to a matched control group [n=39]. A negative control consisting of 17 females was also included. All subjects had two or more children. Blood samples were collected and the ELISA technique was applied to detect antisperm antibodies. Also, the levels of lead in blood and mercury in urine were determined as biological indices of exposure. Antisperm antibodies were detected in 90% of workers exposed to lead with the predominance of the IgG type and 84.6% of workers exposed to mercury with the predominance of the IgM type. Although the results did not correlate with the biological indices of exposure, it seems advisable to use the detection of sperm antibodies of sera of workers exposed to metals as a biological monitoring tool

Immunochemical characterization and antifertility effect of an androgen independent antigen of spermatozoa identified by a monoclonal antibody (D7G3).

Monoclonal antibody, (mAb) D7G3, directed to human spermatozoa and cross-reactive with mouse and rat spermatozoa was found to be a sperm agglutinating antibody. It reacted with antigen present on the post acrosomal region of human spermatozoa and acrosomal region of mouse spermatozoa. The antigen reacting with mAb D7G3 was localized in mouse testicular germ cells and Sertoli cells. It was also localized in the epithelium and spermatozoa of the caput, corpus and cauda epididymides. In addition, the antigen was detected in the epithelial cells and secretions of the prostate and seminal vesicle. The mAb D7G3 reacted with a band of molecular mass 16 kDa in testicular and epididymal sperm and protein preparations of ventral prostate. In the seminal vesicle, an additional band of molecular mass 36 kDa was also identified. The effect of castration on the expression of proteins was studied in the rat. Immunohistochemical localization and Western blotting experiments showed that the antigens identified by the mAb D7G3 was detectable in the epididymis, ventral prostate and seminal vesicle after two weeks of castration. In the seminal vesicle, three additional bands were identified by mAb D7G3 following castration. The expression of this antigen throughout spermatogenesis and sperm maturation indicated that it may have an important biological function. A polyclonal antiserum directed to the 16 kDa mouse epididymal sperm protein was raised in rabbits. Passive immunization of female mice with this antibody caused a significant reduction in fertility only if administered 24 hours prior to mating, indicating that the antibody inhibited a pre-fertilization event by inhibiting sperm function.