Antisperm Antibodies In Seminal Plasma Research Articles

The prognostic value of seminal anti-sperm antibodies screening in men prepared for ICSI: a call to change the current antibody-directed viewpoint of sperm autoimmunity testing.

Aims:We aimed to prospectively evaluate the prognostic value of seminal anti-sperm antibodies (ASA) screening in couples prepared for intracytoplasmic sperm injection (ICSI).Methods:A prospective comparative study was conducted, including 184 non-azoospermic infertile men scheduled for ICSI. For each patient, detection of ASA in seminal plasma using the ELISA technique was done and its relation to standard semen parameters and ICSI outcomes, including fertilization, embryo development, and pregnancy rates, was analyzed.Results:Sperm count, motility, and morphology were negatively affected by existence of seminal ASA (p = 0.012, 0.006, and 0.011, respectively). However, no statistically significant difference was detected between patients with positive and negative seminal ASA regarding the median values of fertilization (56.3% versus 66.7%, p = 0.091), percentage of couples with grade A embryo development (91.4% versus 89.9%, p = 0.520), and pregnancy rates (31.4% versus 32.2%, p = 0.98) after ICSI.Conclusion:ICSI seems able to overcome the problem of ASA in semen. The routine screening of ASA in men prepared for ICSI has no additional prognostic value and cannot be recommended for the time being, until more specific antigen-concerned testing can be developed.

EFFECT OF α–CHYMOTRYPSIN AND GELATIN SUPPLEMENTATION IN RABBIT SEMEN EXTENDER ON ANTI-SPERM ANTIBODYS AND FERTILITY

Three experiments were carried out to study the effects of α-chymotrypsin with gelatin on anti-sperm antibodies (ASAs) in rabbit's semen and fertilization rate. In the first experiment, all rabbits males (20 bucks) were tested for the presence of ASAs in seminal plasma. The secondexperiment, aimed to define the effect of different levels of α-chymotrypsin supplementation (0, 100, 200 and 400 mg/100ml extender) with gelatin in tris-based extender in presence ASAs on progressive sperm motility%, live and abnormal spermatozoa%. In the third experiment, does were inseminated with semen diluted in tris-based extender (1:3) as described in experiment II. Insemination was carried out using 80 does divided into five equal experimental groups (16 does/group) and inseminated artificially by diluted semen containing 50x106 spermatozoa. Kindling rate and litter size were calculated. Results showed significant (P 1/10) as compared with ASAs-negative ejaculates (titer<1/10). Semen ejaculates positive-ASAs supplemented with gelatin and α-chymotrypsin were exhibited ASAs titer 1/10 in seminal plasma that represents a significant change (P<0.05) compared with control group (C). Most amino acids were significantly (P<0.05) increased by the inclusion of gelatin alone or gelatin plus α-chymotrypsin. Supplementation of gelatin and α-chymotrypsin significantly (P<0.05) decreased ammonia in semen, ALT and AST activities in seminal plasma, TBARS concentration, sperm abnormalities, dead spermatozoa, glutathione and superoxide dismutase, while, increased percentages of progressive linear motility as compared to the control. Kindling rate was improved as compared with the control group. In addition, the total and live litter size and bunny weights at birth were considerably higher for supplemented groups α-chymotrypsin and gelatin than those of the control group. The levels of α-chymotrypsin 200 and 400 mg/100 ml extender with gelatin represented the same significant differences with other experimental groups. In conclusion, supplementation of rabbit semen with gelatin and α- chymotrypsin not only enhanced the semen quality but it also improved anti-sperm antibodies status and reproductive performance.

Relationship of Naturally Occurring Antisperm Antibodies in Blood Serum and Seminal Plasma of Cattle Bulls with Sperm Function and Fertility Tests

The study was planned with an objective to assess the level of antisperm antibodies (ASA) in the blood serum and seminal plasma of breeding cow bulls and their relationship with sperm function and fertility tests. ASA was analyzed in blood serum and seminal plasma by SpermMar test, Immuno peroxidase assay (IPA) and Enzyme linked immunoabsorbant assay (ELISA). In SpermMar test, about 54% bulls were with >40% IgG in blood serum against sperm surface antigens, whereas none of the bulls were with >10% IgG in seminal plasma. More than 20% and >10% IgA against sperm surface antigens were detected in the blood serum and seminal plasma of 65.8% and 37% bulls, respectively. Out of 26 bulls, seminal plasma of 21 bulls reacted with spermatozoa both in IPA and IgA latex particles and that of only 12 bulls reacted with IgG. In IPA, about 50% of the bulls had >40% ASA against head surface antigens, whereas, there were 23% bulls with >10% ASA in seminal plasma. Also ELISA indicated a higher antibody titre in blood serum (3200 - 6400) and seminal plasma (40 - 80) of 50% and 42% bulls, respectively. There were 11 bulls with low values of HOST/in vitro acrosome reaction/cervical mucus penetration assay and higher level of either serum or seminal plasma ASA. Our study revealed that a significant level of ASA in serum or seminal plasma may have effect on the fertility of bulls by affecting the sperm function.

The influence of antisperm antibodies, intratesticular haemodynamics and the surgical approach to varicocelectomy on seminal variables

ObjectiveTo evaluate the effect of antisperm antibodies (ASAs), hormonal levels, intratesticular haemodynamics and the surgical approach on the outcomes of varicocelectomy in infertile men, as assessed by seminal variables. Patients and methodsIn a prospective case-series study, 82 infertile men with varicocele (35 left and 47 bilateral) were evaluated. The preoperative assessment included a physical examination, semen analysis, assessment of ASAs in seminal plasma, hormonal levels (follicle-stimulating hormone (FSH), luteinising hormone and testosterone), and scrotal colour Doppler ultrasonography (CDUS) to measure the peak systolic velocity (PSV), end diastolic velocity (EDV), resistive index (RI) and pulsatility index. Patients were scheduled for varicocelectomy, with high ligation (Palomo) used in 40 patients (18, 45%, with left and 22, 55%, with bilateral varicocele), or an inguinal approach (Ivanissivich) with loupe magnification used in 42 (17, 40%, with left and 25, 60%, with bilateral varicocele). The men were reassessed at ⩾3months after surgery and according to the improvement in seminal variables (expressed as a ⩾50% increase in total motile sperm count, TMSC), patients were further categorised into ‘improved’ or ‘unimproved’. Binary logistic regression analysis was used to investigate the predictors of improvement. ResultsBefore surgery the ASAs were positive in 17 men (21%). There was no significant difference between the right and left sides in intratesticular haemodynamics. The TMSC was improved in 52 (63%) patients who had a significant improvement in the haemodynamic variables. Intratesticular haemodynamics, serum FSH and testosterone levels differed significantly between the improved and unimproved patients. Positivity for ASAs, the surgical approach and laterality of the varicocele were not significantly different, although the ASA-positive cases were characterised by a significant decrease in motility. Logistic regression analysis showed that the EDV, PSV, FSH, testosterone level and bilateral testicular volume (BTV) were significant predictors of improvement. ConclusionPositivity for ASAs is not a predictor of the outcome after varicocelectomy but affects only the motile fraction in positive cases, despite the improvement in other seminal variables and testicular haemodynamics, and regardless of the surgical approach. The EDV, PSV, FSH, testosterone and BTV were significant predictors of a successful outcome.

Are Antisperm Antibodies Really Associated with Proven Chronic Inflammatory and Infectious Diseases of the Male Reproductive Tract?

It is debated whether chronic urogenital inflammations and infections may trigger the formation of antisperm antibodies (ASA) in semen. To evaluate the formation of ASA in defined chronic inflammatory and infectious diseases of the male reproductive tract (MRT). Three hundred sixty-five patients retrospectively enrolled in a single center were categorized as having National Institutes of Health (NIH) category II chronic prostatitis (n=38), NIH category IIIa chronic prostatitis/chronic pelvic pain syndrome (CP/CPPS) (n=59), NIH category IIIb CP/CPPS (n=213), chronic epididymitis (n=34), and chronic urethritis (n=21). Forty-five age-matched men served as controls. All subjects underwent microbiologic and cytologic analysis for common bacteria, yeasts, and mycoplasma using the four-glass test. Urine samples, ejaculates, and urethral swabs were analyzed with polymerase chain reaction (PCR) for Chlamydia trachomatis and Neisseria gonorrhea. Semen analysis followed World Health Organization (WHO) standards. ASA in seminal plasma were analyzed using the mixed agglutination reaction (MAR) test. The overall positive detection rate of clinically significant levels (> or = 50% of spermatozoa coated by ASA) of IgG and IgA antibodies was 1.8% and 0.8%, respectively, in the patient group. No clinically significant levels of ASA were detected in the control group, and no statistically significant difference was observed between controls and patients (IgG, p=1.0; IgA, p=1.0). No difference was found between the different inflammatory and infectious diseases and the control group in the detection rate of ASA, even when the cut-point value was lowered to > or = 1% (IgG, p=0.4; IgA, p=0.3). Moreover, in one selected subgroup of patients (n=26) with persistent increased inflammatory parameters (peroxidase-positive leukocytes [PPL] > or = 1 x 10(6)/ml and elastase > or = 230 ng/ml), no significant difference in the levels of ASA was observed compared with the controls (IgG, p=0.1; IgA, p=0.8). There is no association between chronic inflammatory or infectious diseases of the MRT and the presence of ASA in semen.

Detection of antisperm antibodies in seminal plasma by flow cytometry: comparison with the indirect immunobead binding test

Objective: To compare flow cytometry with the established indirect immunobead binding test (IBT) for the detection of antisperm antibodies in seminal plasma. Design: A prospective, comparative study. Setting: University-based and rology unit. Patient(s): One hundred and fifty-eight men with suspected male factor subfertility. Intervention(s): Seminal plasma samples were incubated with antisperm antibody-negative donor sperm. Surface-bound antibody was detected with fluorescence-labeled antihuman antibody in the flow cytometry assay or with immunobead-labeled antihuman antibody in the IBT. Main Outcome Measure(s): The percentage of sperm that tested positive for surface-bound antibody was determined in the two assays. Seminal plasma was antisperm antibody-positive when ≥20% of the sperm were antibody-bound, and clinically significant levels were present when ≥50% of the sperm were antibody-bound. Result(s): Of 71 samples that were negative by the IMT, 66 (93%) also were negative by flow cytometry. Of 63 samples that had ≥50% immunobead binding, 55 had equivalent results by flow cytometry. Overall statistical analysis showed a good correlation between the two assays. Conclusion(s): There is a good correlation between the indirect IBT and indirect flow cytometry for the detection of antisperm antibodies in seminal plasma.

Seminal fluid antisperm antibodies measured by direct flow cytometry do not correlate with those measured by indirect flow cytometry, the indirect immunobead test, and the indirect mixed antiglobulin reaction

To compare the direct flow cytometric (FCM) measurement of sperm-bound antibodies with the indirect FCM measurement of unbound antisperm antibodies in seminal plasma of the same ejaculates. To compare the FCM measurements with the indirect mixed antiglobulin reaction (MAR) and the indirect immunobead test (IBT) performed on the same seminal plasmas. University hospital-based infertility service. Eleven infertile men with a positive immunoglobulin (Ig) G direct MAR test and antibody positivity on the direct FCM measurements. Four control patients with a negative IgG-direct MAR test. The proportion of spermatozoa positive for IgG and IgA antibodies as measured by various tests. The direct and indirect FCM measurements for sperm-bound antibodies from seminal plasma did not correlate with each other. In general, IgG antibodies were not detectable by indirect FCM whereas IgA were detected in lower proportions than by direct FCM measurements. Weak correlation was observed between the indirect FCM, indirect MAR, and indirect IBT measurements. Nearly all of the IgG and some of the IgA antisperm antibodies in seminal fluid bind to spermatozoa. Thus, indirect tests to measure antisperm antibodies from seminal plasma are likely to miss the presence of antisperm IgG antibodies while detecting some cases of IgA antibodies.

Nonspermatozoal cells in semen: relationship to other semen parameters and fertility status of the couple.

In semen samples from 94 men from infertile couples and 42 control patients proven to be fertile the occurrence of different kinds of nonspermatozoal cells (NSC) was examined. The findings were evaluated in relation to other seminal parameters and to the fertility status of the couple. The ratios of NSC to sperm and primary spermatocytes to spermatids revealed increasing values with decreasing sperm counts. Especially in a group where the semen samples contained > or = 6 x 10(6) motile sperm per mL and no infertility factor was detectable in the woman, the concentration of leukocytes was high, with three of eight patients having > or = 10(6) leukocytes per mL. A high proportion of abnormal sperm seemed to occur more frequently where the cause of infertility could be found in the man. A high number of leukocytes was neither significantly associated to an increased proportion of abnormal sperm nor to sperm motility evaluated by recording the percentage of motile sperm and the 1-h migration distance in sodium hyaluronate medium. No correlation between concentration of lymphocytes and presence of antisperm antibodies in seminal plasma was detectable.

Antisperm antibodies in seminal plasma of spinal cord-injured men.

Central to the problem of reproductive rehabilitation of spinal cord-injured men treated by assisted ejaculatory techniques is the consistent observation of deficient semen quality. Most studies have reported asthenospermia despite the presence of normal sperm concentration in most men undergoing these procedures. To date little attention has been given to the incidence and relevance of sperm autoimmunity in this group. In 9 anejaculatory spinal cord-injured men, electroejaculation was performed. Antegrade ejaculates were obtained in 7 men and analyzed. Mean sperm antegrade concentration was 74.4 +/- 113 x 10(6)/mL with a mean motile sperm concentration of 28.6 +/- 54.0 x 10(6)/mL. Enzyme-linked immunosorbent assay (ELISA)-determined antisperm antibody response was positive in the seminal plasma of 5 of 7 patients. Because of the disproportionately high incidence of an immunologic factor in men with neurogenic infertility, sperm autoimmunity should be considered among the important causes underlying their seminal dysfunction.

Seminal plasma anti-sperm antibodies and IVF: the effect of semen sample collection into 50% serum

A retrospective analysis was carried out which studied IVF results for couples in whom the male partner had anti-sperm antibodies in seminal plasma, as judged by a positive MAR test. Of the 59 couples studied over a total of 113 cycles, 30 had IVF semen samples collected into sterile, dry pots. Thirty-eight of the couples had samples collected into medium containing 50% serum, and nine couples underwent separate treatment cycles with samples collected both dry and into 50% serum. The results demonstrate that the addition of serum to sample collection pots significantly improves the oocyte fertilization rate and is followed by a greater chance of conception for these couples.

Antisperm antibodies and sperm motility: a study using timed exposure photomicrography.

This study examined the effect of antisperm antibodies on sperm motility. Antisperm antibodies present in seminal plasmas with different sperm agglutinating titres were transferred passively to normal donor sperm, and the effects on movement characteristics (velocity of forward progression, amplitude of lateral head displacement, percentage progressive motility and percentage non-progressive motility) were analysed using timed exposure photomicrography. There was no significant association between sperm movement characteristics and the presence of titre of antisperm antibodies in seminal plasma. Furthermore, no differences were detected between those samples that possessed sperm agglutinating versus sperm immobilizing activity. These findings do not support the common belief that antisperm antibodies are a cause of poor sperm motility in semen.

Mixed erythrocyte-spermatozoa antiglobulin reaction (MAR test) for IgA antisperm antibodies in subfertile males

A mixed antiglobulin reaction (MAR test), using antibody-coated red cells and spermatozoa, has been used to detect the presence of either IgA or IgG on he surface of the spermatozoa. Group O Rh-positive red cells were sensitized with serum containing anti-D, which was partly IgA, though mainly IgG. Spermatozoa were added to the sensitized red cells and tested either with anti-IgA or anti-IgG. MAR (IgA) test results were negative or doubtful in 42 (44%) of 94 samples with positive or strongly positive MAR (IgG) tests. Positive MAR (IgA) tests showed a highly significant correlation with the presence of antisperm antibodies in seminal plasma. Significantly impaired sperm penetration of cervical mucus was demonstrated for 15 patients, 12 of whom had positive MAR (IgA) tests, whereas good sperm penetration was observed for 5 patients with negative or dubious MAR (IgA) tests; all 20 patients had strongly positive MAR (IgG) tests and positive serum antisperm antibody tests.

Vasectomy: Consequences of Autoimmunity to Sperm Antigens

Data from studies examining the effects of vasectomy in a large number of nonhuman primates vasectomized for periods ranging up to 14 years are summarized, and these findings and speculations are used as a framework with which to review the subject of autoimmunity and vasectomy. Attention is directed to autoimmunity to sperm antigens following vasectomy (factors affecting antisperm antibody levels, characteristics of circulating antisperm antibodies, antisperm antibodies in seminal plasma, and cellular immunity following vasectomy), and immunopathology of antisperm autoimmunity (local effects on the male reproductive tract and systemic effects on the male reproductive tract). The 6 hypotheses that have been advanced to explain individual variations in dynamics and types of antisperm antibodies produced following vasectomy are reviewed. 3 tests are commonly used to detect free antisperm antibodies after vasectomy: 1) the spermagglutination test; 2) the sperm immobilization test; and 3) the immunofluorescence test. Spermagglutinating (SA) antibodies, the most common type of antibody produced after vasectomy, occur in approximately 2/3 of vasectomized men and in a majority of vasectomized rhesus monkeys. Sperm-immobilizing (SI) antibodies are also produced in a large percentage (40%) of vasectomized men and rhesus monkeys. About 30% of vasectomized men also have antiprotamine antibodies.