Anti-sperm antibodies detection by a modified MAR test: Towards a better definition of its indications.

RESEARCH QUESTION: Anti-sperm antibodies (ASA) have been shown to reduce male fertility but consensus about the precise situations in which tests should be carried out are lacking. In infertility investigations, should the mixed antiglobulin reaction (MAR) test be a first-line test? Should it be carried out systematically before assisted reproductive technology (ART)? What are the risk factors for ASA? DESIGN: All infertile patients (n = 1364) were tested with SpermMar (modified MAR test) between July 2013 and June 2017. Intra-patient variability of the MAR test was also assesed by comparing two tests within the same year in selected patients (n = 101). RESULTS: The main factor that influenced the percentage of ASA was the presence or absence of sperm agglutination. In the presence of agglutinations, 27 out of 72 (37.5%) patients were positive for ASA compared with 33 out of 1292 (2.6%) in the absence of agglutinations (P < 0.0001). When one risk factor was present (spontaneous sperm agglutination, history of scrotal trauma or inguinal surgery), 33 out of 179 (18.44%) tests were positive for ASA (≥50% coated spermatozoa), whereas only 27 out of 1242 (2.2%) were positive when no risk factor was present (P < 0.0001). CONCLUSIONS: ASA detection should not be systematically recommended in investigations of fertility status and before ART but reserved for when sperm agglutination is found during conventional sperm examination, or if the patient has a history of scrotal trauma or has undergone inguinal surgery.

Antisperm antibodies in repeat-breeding cows: Frequency, detection and validation of threshold levels employing sperm immobilization, sperm agglutination and immunoperoxidase assay.

Antisperm antibodies have been found in repeat-breeding(RB) cows, and those causing agglutination and/or immobilization of sperm are considered to be closely related to unexplained infertility. However, a standard protocol for identifying antisperm antibodies (ASA) in cattle is not validated. Therefore, an investigation was undertaken to evaluate sperm immobilization (SIT), sperm agglutination (SAT) and immunoperoxidase (IPT)assays for detection of ASA in serum and their respective threshold levels for confirmation. Animals (heifers, normally breeding, repeat-breeding and pregnant animals) that were free from IBR, brucellosis and uterine infections (screened by clinical examination) were included in the study. Sperm agglutinating, sperm immobilizing and antisperm antibodies evaluated by respective assay were significantly higher (p < .05) in RB cows compared to other groups. The SIT assay was able to identify 61% of RB caused by ASA, more than those employing SAT and IPT. Furthermore, a dilution rate of 1:5 and 1:80 (confirms 59.0 and 57.0% RB+ve)were sufficient to diagnose ASA by SAT and IPT, respectively. Results indicate the presence of __12.6% clumped spermatozoa and __ 2.6%(cut-off value) peroxidase-positive spermatozoa at 1:5 and 1:80 dilutions diagnosed with SAT and IPT, respectively, may be considered as repeaters arising out of ASA. Furthermore, study also showed the presence of lower incidence of ASA positivity in other groups of animals (heifer

Methods for direct and indirect antisperm antibody testing.

Antisperm antibodies (ASA) are one well-known cause of refractory infertility in both males and females. In females, a sperm immobilization test, which detects sperm-immobilizing antibodies indirectly in the patient's serum, requires complement for the reaction and thus seems to be a more specific immunological reaction. In males, an immunobead test or a mixed antiglobulin reaction test, which detects ASA directly on the sperm surface, is a screening test because of the nonspecific reaction.

An isoimmune response to human sperm clathrin in an infertile woman with systemic lupus erythematosus.

We have employed a proteomic approach to study the immune response to human sperm in an infertile female patient suffering from systemic lupus erythematosus (SLE). Human sperm antigenic extracts were resolved by means of two-dimensional electrophoresis and electroblotted onto nitrocellulose membranes. The membranes were incubated with serum from the SLE patient. Sperm antigens that were reactive to polyclonal antibodies were next visualized on X-ray film, using the enhanced chemiluminescence (ECL). Three spots corresponding to the positions of sperm immunoreactive antigens on a nitrocellulose membrane were localized in a silver stained gel and subjected to mass spectrometry. A database search of the sequences recognized by the analyzed SLE serum revealed its homology to the clathrin heavy chain (CHC). Further analysis revealed that anti-CHC antibody reacted with multiple sperm antigenic determinants, resolved by either one- or two-dimensional electrophoresis. When studied by immunofluorescence, we demonstrated anti-CHC antibody reactivity with the sperm tail tip (corresponding to the sperm agglutination pattern), also with the principal piece and with cytoplasmic droplets around the sperm midpiece. Live sperm clearly exhibited reactivity with the midpiece. This study demonstrates clathrin heavy chain on human sperm using serum of an infertile individual with a concomitant autoimmune disease.

[Local sperm antibodies and general levels of immunoglobulins in ovulatory cervical mucus]. / Lokální protilátky proti spermiím a celkové hladiny imunoglobulinu v ovulacním cervikálním hlenu.

OBJECTIVE: The aim of our preliminary study was to compare the levels of total local sIgA and IgG with activity of detected sperm antibodies in ovulatory cervical mucus (OCM). SETTING: Department of Gyneacology and Obstetrics, Medical School and Faculty Hospital, Charles University, Plzen. METHODS: We screened samples of OCM from 12 pacients aged 26-31 (29,6 years on average) by radial immunodifusion (RID) to determine s IgA and IgG. Indirect MAR test was used for detection of spermagglutinationg antibodies. RESULTS: We found out by RID the average concentration of sIgA in OCM 567,84 mg/l (0 -1250,47) and the average concentration of IgG in OCM 23,57 mg/l (8,74-47,99). Antibody activity against sperm cells dominates in IgA with 6 pacients, in IgA with 1 patient, in IgA and IgG together with 1 infertile woman and in IgA and IgM isotypes together with 1 patient. No local sperm antibodies were determined with 3 patients. CONCLUSION: We proved the hypothesis, that the levels of spermagglutinating antibodies do not correlate with findings of total sIgA and IgG in OCM with our patients.

[Isolation and immunology identification of spermagglutinating antibodies from human serum]. / Izolace a imunologická identifikace spermaglutinacní protilátky z lidského séra.

OBJECTIVE: Isolation of spermagglutinating antibodies and their assesment. DESIGN: Retrospective study. SETTING: Special consulting for reptoduction immunology, Department of Obstetrics and Gynecology, Charles University and Faculty Hospital, Plzen. METHODS: Fractionation of serum samples by liquid exclusion chromatography, examination of full sera and their chromatographic fractions by Friberg teste (Tray Agglutination Test--TAT), indirect antiimmunoglobulin reaction test (i-MAR test) and by supplementar radial immunodiffusiona (RID). RESULTS: Isolation of spermagglutinating fractions of antisperm antibodies positive sera preserved spermagglutinating aktivity and confirmed great spermagglutinating potential of IgM. CONCLUSION: According to assesment of the presence of IgG and IgG we reported possible states of immunisation: actual immunisation with IgM activity, perpetual stimulation (IgG and IgM) and, finaly, anamnestic titres in IgG. These findings can help us to choose an optimal way of treatment. Excluding gel chromatography is suitable method for serum proteins fractionation, but not their identification--presence of antisperm antibodies does not affect the chromatographic spectrum, nor the RID patterns.

Sperm antibodies, intra-acrosomal sperm proteins, and cytokines in semen in men from infertile couples.

PROBLEM: The aim of this study was to investigate seminal sperm-agglutinating antibodies, intra-acrosomal proteins, sperm head abnormalities, and cytokines (IL-1beta, IL-2, IL-4, IL-5, IL-6, IL-8, IL-10, IL-12p70 TNF-alpha, and IFN-gamma) in men from infertile couples. METHOD OF STUDY: The direct mixed anti-immunoglobulin reaction test for IgG, IgA, and IgE in semen, and immunocytochemical method using monoclonal antibodies and indirect immunofluorescence for the examination of intra-acrosomal proteins in the spermatozoa were used. Cytokines in seminal plasma were determined by multiplex immunoanalytic xMAP (LUMINEX) technology. RESULTS: Sperm-agglutinating antibodies, IgG and IgA, in seminal plasma were found to be more in asthenospermatic and oligoasthenospermatic men than in normospermatic men. Sperm head pathology and very low amounts of acrosomal proteins were frequently detected in pathologic semen samples. Cytokine levels defined as 'high' (based on the 75 percentile for each cytokine in all groups) were obtained especially for IL-8, IL-5, IL-6, and IL-10. The high cellularity in semen was correlated with higher IL-5. CONCLUSION: Immunologic cause of male infertility is a very important risk factor in the pathogenesis of sperm cells. Sperm autoantibodies and the presence of intra-acrosomal factors must be studied together, cytokines according to accessory cellularity in the semen.

Experimental immunological infertility effect of anti-GAPDH-2 antibodies on the fertility of female mice.

OBJECTIVE: To examine the relationship between an antibody against GAPDH-2, a sperm-specific protein, and infertility of female mice. DESIGN: Basic research. SETTING: National Research Institute for Family Planning Beijing, World Health Organization Collaboration Center of Human Reproduction. ANIMAL(S): New Zealand rabbit, NIH and ICR mice. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): Enzyme-linked immunoabsorbent assay, Western blot and indirect immunostaining assays, standard fertility assay, and sperm agglutination assay. RESULT(S): Antibodies against the full-length GAPDH-2 were raised. Its specificity was assessed by immunoblotting and indirect immunostaining assays. The antibody immunoreacted with human sperm GAPDH-2 and the mouse homolog GAPDS but did not cross-react with GAPDH. Treatment of female mice with IP injection of anti-GAPDH-2 serum significantly reduced their fertility. Anti-GAPDH-2 serum caused the agglutination of normal mice sperm in vitro. The anti-GAPDH-2 antibody was detectable in the sera and uterine fluid of the mice immunized with GAPDH-2. CONCLUSION(S): These results show that GAPDH-2 should be further evaluated as a promising candidate in the development of an antifertility immunogen. Detecting anti-GAPDH-2 antibodies in the bodily fluid of subjects afflicted with indeterminate infertility may be a new diagnostic index.

Expression of a recombinant human sperm-agglutinating mini-antibody in tobacco (Nicotiana tabacum L.).

The murine monoclonal antibody (mAB) S19 recognizes an N-linked carbohydrate antigen designated sperm agglutination antigen-1 (SAGA1) located on the membrane protein CD52. This antigen is added to the sperm surface during epididymal maturation. Binding of the S19 mAB to SAGA-1 causes the rapid agglutination of sperm and blocks pre-fertilization events. Previous studies indicated that the S19 mAB may be a potential specific spermicidal agent (termed a spermistatic) capable of replacing current spermicidal products that contain harsh detergents with harmful side effects. The nucleotide sequences encoding the heavy (H) and light (L) chains of the S19 antibody were cloned. A chimeric gene was constructed using the nucleotide sequences encoding the variable regions of both the H and L chains, and this gene (scFv1 9) was expressed in transgenic tobacco (Nicotiana tabacum L.) to produce a recombinant anti-sperm antibody (RASA). Highest levels of RASA expression were observed in BY-2 plant cell suspension cultures and regenerated N. tabacum cv. Xanthi plants transformant in which the RASA coding sequences were expressed under the control of the Cauliflower Mosaic Virus 35S promoter containing a double-enhancer sequence (2X CaMV 35S). Subsequent modifications of the transgene including the addition of a 5'-untranslated sequence from the tobacco etch virus (TEV leader sequence), N-terminal fusion of the coding region with an endoplasmic reticulum targeting signal of patatin (pat) and C-terminal fusion with the endoplasmic reticulum retention signal peptide KDEL showed further enhancement of RASA expression. The plant-expressed RASA formed intrachain disulfide bonds and was primarily soluble in the cytoplasmic fraction of the cells. Introduction of a poly-histidine (6xHIS) tag in the recombinant RASA protein allowed for rapid purification of the recombinant protein using Ni-NTA chromatography. Optimization of scale-up production and purification of this plant-derived recombinant protein should provide large quantities of an inexpensive spermistatic plantibody.

[Inhibin B and intraacrosomal proteins in men from the couples with fertility disorders]. / Inhibin B a intraakrozomální proteiny u muzu z páru s poruchou fertility.

THE AIM: To monitor the basic andrologic and immunologic sperm factors and the levels of inhibin B in serum and in seminal plasma in men from the couples with infertility disorders. SETTING: Department of Gynecology and Obstetrics, Medical School, Charles University and University Hospital, Plzen, Institute of Molecular Genetics, AV CR, Prague, Institute of Clinical Immunology and Allergology, LF UK a FN, Plzen. METHODS: We used conventional methods for estimation of sperm quality according to WHO and we detected the intra-acrosomal proteins by monoclonal antibodies (Hs8 and Hs14, immunofluorescent method), spermantibodies by direct mixed antiimunoglobulin reaction (MAR) test, and we examined inhibin B in serum (< or =400 pg/ml= A) and in seminal plasma (< or = 600 pg/ml= N) by ELISA in 355 men aged 21-52 years (ø 34 years) with normal levels of FSH, LH and testosterone. The control group was created by 56 health sperm donors. RESULTS: We found 65% normospermatics in the group of 355 patients, 34.9% men with various kind of pathologies. Predominance of spermagglutinating antibodies was found in 15.77% in IgG, in 19.44% in IgA, in 8.44% in IgA and IgG together. Normal intraacrosomal proteins were reached in 74.65% for Hs8, in 20.85% pathologic, in 86.2% normal findings for Hs14, in 4.23% pathologic. The immunological results in control group were completaly negative. Pathological levels of inhibin B in seminal plasma was found in 37.2% (152 men), in 25% in serum, and in 5.6% in serum and in seminal plasma together. In 54.7% of patients we found physiological levels of inhibin B in both biological fluids. We also compared physiological 109/152 (71.71%), and pathological spermiogrammes 43/152 (28.29%) with abnormal levels of inhibin B in seminal plasma, with intraacrosomal proteins to levels of inhibin B in serum. Our detailed study shows high interidividual results, which must be studied in complex with diagnosis of decreased fertility in man. CONCLUSION: Andrologic and immunologic analysis in the group of 355 men showed normal parameters of spermiogrammes in 231 patients (65%), in the rest of men the immunologic profil was in various parts pathologic. Only 105 men have got excellent spermiogrammes. Inhibin B as hormon regulates in back the secretion of FSH, and serves as good indicator in male reproductive failures.

[Ovulatory mucus and its pH, arborization and spermagglutination antibodies in women with fertility disorders]. / Ovulacní hlen a jeho pH, arborizace a spermaglutinacní protilátky u pacientek s poruchou plodnosti.

OBJECTIVE: We studied pH of ovulatory mucus, sperm penetration through capillary filled with ovulatory mucus in one hour and presence of local spermagglutinating antibodies. METHODS: We measured pH, arborization and Kremer test in ovulatory mucus in 127 women with fertility disorder. Indirect mixed antiimunoglobulin reaction test (i-MAR-test for IgG, IgA, IgM and IgE) was used for detection of spermagglutinating antibodies. RESULTS: pH 7.4-9.6 (physiological limit) was found in 94/127 women (74%), pH < 7.4 in 33 women (26%). 60% of 94 patients with physiological pH had Kremer's test above 2cm/hour (normal sperm penetration in ovulatory mucus), in 40% of them reduction of sperm penetration (< 2cm/hour, swelling, shaking, cytotoxicity) was seen. Sperm antibodies in ovulatory mucus in 24% patients with pH < 7.4 and 22% patients with physiological pH were studied. In 111 patients with regular menstrual cycle a classical arborization was found in 81%, in 14% was not proved. In 16 patients with irregular menstrual cycle classical arborization was observed in 11 of them. Local sperm antibodies were detected only in 13% of the total count of patients, it means in 17 patients. Their ovulatory mucus showed classical arborization. 30 healthy fertile women created the control group, only one secretion had pathological findings in all studied parameters owing to latent mycotic infection. SUMMARY: Pathological pH of ovulatory mucus, hormonal dysbalance, and presence of local spermagglutinating antibodies evidently influence penetration of sperm cells through cervix uteri. Otherwise pathological microbial vaginal environment can start a cross reaction with the surface microbes and sperm epitopes. One sign of ovulation, arborization of cervical ovulatory mucus, is not connected directly with the presence of local sperm antibodies, but insufficient estrogen influence is a sign of the reduced immunosuppression in cervix area.

Isolation of a sperm-agglutinating factor from Staphylococcus aureus isolated from a woman with unexplained infertility.

Spermatozoal agglutinating factor (SAF) that agglutinates human spermatozoa has been isolated from Staphylococcus aureus. By scanning electron microscopy, Staphylococcus aureus adherence was observed on sperm tails.

[The incidence of agglutination and its influence on sperm quality and fertility of boar semen]. / Untersuchungen über das Vorkommen von Agglutinationen und deren Einfluss auf die Spermaqualität und Fertilität beim Ebersperma.

The aim of this study was to examine the incidence of sperm agglutinations and their relationships with sperm quality and fertility. Semen samples of 40 boars of an AI-station were investigated. Nineteen of the 40 investigated boars showed a constantly low (< 10% agglutinated sperm), 3 an intermediate (10-20%) and 6 boars a high level (> 20%) of agglutination in raw semen. The degree of agglutination in sperm samples of 12 boars varied distinctly during the investigation period. During summer more (P < 0.05) agglutinated sperm were observed (11.0 +/- 11.6%) than during winter (6.2 +/- 7.3%). There was no association between bacterial contamination and incidence of agglutinations (P > 0.05). After dilution in extender the percentage of agglutinated sperm decreased from 6.2 +/- 7.3% to 1.1 +/- 1.4% (P < 0.0001). Twenty-four hours after dilution the percentage of progressively motile sperm was 7.4% lower (P < 0.05) in ejaculates with an initially high degree of agglutination (> 20% agglutinated sperm) compared to samples with an initially low degree of agglutinated sperm (< 10%). Plasma membrane integrity, mitochondrial membrane potential, acrosome reaction and chromatin structure were independent (P > 0.05) from the level of agglutination. Fertility data did not differ (P > 0.05) between boars with low and high numbers of agglutinated sperm in raw semen. The results show that there are individual, ejaculatory and seasonal variations in the incidence and degree of agglutination. Agglutinations have a negative effect on motility of sperm and disappear to a large extent after dilution in sperm extender. They have no negative consequences on fertility.

Sperm granuloma and sperm agglutination in a dog with asthenozoospermia.

A mongrel dog, aged 2 years, was found to have only a small number of sperm, immobilization of all sperm, and many sperm agglutinations in its ejaculates, and scrotal palpation revealed a small nodule in the left cauda epididymis. Addition of the dog's seminal plasma or serum to the semen of 2 normal dogs caused immobilization and agglutination of their sperm. Histological examination showed that the nodule was a sperm granuloma. Many lymphocytes were seen in the stroma around the sperm granuloma. Anti-sperm antibodies are presumed to be present in the semen and serum of the asthenozoospermic dog.

Effect of immunization with synthetic peptide corresponding to region 1-17 of human seminal plasma inhibin on fertility of male rats.

Immunization of adult male rats with a synthetic peptide corresponding to the region 1-17 of human seminal plasma inhibin (hSPI) resulted in agglutination of epididymal sperm, severely affecting the fertility of the animals (75% reduction in fertility as compared to control). This effect was found to be dependent on the antibody titer to hSPI. There was a significant rise in circulating follicle-stimulating hormone levels, with luteinizing hormone and testosterone levels remaining unaffected. The histology of the testes and other reproductive organs revealed that these organs remained unaltered. The N-terminal 1-17 amino acid peptide of hSPI may hold promise as an immunogen for male immunocontraception.

Characterization of a potent sperm-agglutinating monoclonal antibody and its cognate antigens.

OBJECTIVE: To identify sperm antigens that are capable of eliciting infertility-related sperm-agglutinating antibodies. DESIGN: In vitro laboratory experiments. SETTING: University research laboratory. PATIENT(S): Fertile semen donors. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): Sperm agglutination, immunofluorescence localization, and flow cytometric analysis of surface expression of A36 antigens. Antigen analysis by Western immunoblotting. RESULT(S): Monoclonal antibody A36 induced intensive head-to-head, tail-to-tail, and head-to-tail agglutination of motile human spermatozoa. Antigens recognized by A36 were localized on the acrosomal cap and in the principal tail regions of motile, noncapacitated human sperm. Changes in subcellular levels and localization of the A36-recognized epitope occurred after capacitation and acrosomal loss. A36 reacted with a polymorphic series of proteins in Western blots of sperm extracts from humans and various other animal species, including mouse testis extracts. A common 53-kd antigen was recognized by the antibody in the different antigenic preparations. CONCLUSION(S): A mouse antibody to human sperm, monoclonal antibody A36, caused intensive agglutination of noncapacitated human spermatozoa and reacted with antigens on the acrosomal cap and in the principal tail regions. Of the multiple polypeptides that were reactive with the monoclonal antibody in sperm extracts from humans and other animal species, a common 53-kd antigen was recognized.

Changes in epididymal protein anti-agglutinin on ejaculated boar spermatozoa during capacitation in vitro.

This study is a detailed investigation of changes in epididymal protein anti-agglutinin on ejaculated boar spermatozoa during an incubation designed to promote capacitation in vitro. Ejaculated spermatozoa were collected from six mature boars, washed, and incubated to promote capacitation. Sperm samples were subjected to Western blotting-densitometric analyses, flow cytometry after immunostaining and immunocytochemical observation by indirect immunofluorescence. An antiserum to anti-agglutinin was raised in a rabbit by subcutaneous injection of a purified antigen, as described previously (Harayama et al. 1999). Western blotting-densitometric analyses revealed an approximate halving of the amount of sperm-bound anti-agglutinin during the first 45-min incubation, followed by a gradual decrease thereafter. Comparison between immunostained sperm samples by flow cytometry before and after incubation confirmed this decrease in sperm-bound anti-agglutinin during the incubation. Microscopic characterization established that this decrease occurred mainly on the acrosome. Supplementation with seminal plasma (5% or 10%, v/v) attenuated the decrease. These findings are consistent with the conclusion that a large portion of the anti-agglutinin bound to sperm acrosomes is released at an early stage of the capacitation process in vitro.

Biochemical characterization of sperm agglutination antigen-1, a human sperm surface antigen implicated in gamete interactions.

The anti-sperm monoclonal antibody (mAb) S19 was previously demonstrated to agglutinate human spermatozoa, inhibit sperm penetration of cervical mucus, and inhibit sperm-zona pellucida binding. These results implicated the cognate S19 antigen, designated sperm agglutination antigen-1 (SAGA-1), in gamete interactions and identified SAGA-1 as an attractive candidate for immunocontraceptive development. In the present study, evaluation of sperm agglutination with video microscopy showed that the S19 mAb rapidly and completely agglutinated human spermatozoa in a "tangled" pattern of agglutination. One- and two-dimensional immunoblot analyses identified SAGA-1 as a highly acidic, polymorphic sperm protein with an apparent molecular mass of 15-25 kDa and an isoelectric point of 2.5-3.0. Periodate treatment abolished this immunoreactivity, demonstrating that the S19 mAb reacted with a carbohydrate epitope and indicating that SAGA-1 is a glycoprotein. Absence of S19 immunoreactivity in postvasectomy seminal fluid implicated the testis, epididymis, and/or proximal vas deferens in the expression of SAGA-1. In solubility and phase partitioning assays, SAGA-1 was extracted from spermatozoa in Triton X-114 and exhibited the hydrophobic characteristics of integral and glycosylphosphatidyl inositol-anchored membrane proteins. These results identify SAGA-1 as a hydrophobic, highly acidic sperm glycoprotein that is localized on the entire sperm surface and has potential significance as a target for antibodies that inhibit sperm function and gamete interactions.

Acrobeads test as a predictor of fertilization in vitro.

PROBLEM: To determine whether the results of the Acrobeads test, which measures the expression of the complement regulator molecule CD46 on the inner acrosomal membrane following the acrosome reaction, accurately identifies semen specimens that will exhibit reduced or failed fertilization following conventional IVF insemination. METHOD: The Acrobeads test was performed on semen specimens from 97 consecutive patients preparing to undergo an IVF cycle utilizing a standardized insemination protocol. Motile sperm populations were examined at 6 h and 24 h post-isolation for sperm-bead agglutination. Results of the Acrobeads test were compared to that of TRITC-PSA staining in matched specimens to directly measure the spontaneous loss of acrosome content. The percentages of TRITC-PSA-negative sperm were determined in freshly isolated motile populations and in duplicate aliquots incubated 18 to 20 h under sperm capacitating conditions. The relationship between the results of both analyses estimating spontaneous acrosome reactions and the rate of fertilization of metaphase II oocytes was examined. RESULTS: The Acrobeads score did not correlate significantly with the rate of fertilization by insemination at 6 h or at 24 h. The negative predictive value of this test was 21.4%. There was no correlation between the Acrobeads score and the percentage of sperm undergoing a spontaneous acrosome reaction as detected by TRITC-PSA labeling. In contrast, the increment increase in the percentage of spontaneous acrosome reactions as quantified by TRITC-PSA staining was correlated with the fertilization rate. CONCLUSIONS: Contrary to previous reports, our prospective, double-blinded study failed to demonstrate that the Acrobeads test can accurately predict fertilization outcome in IVF. Therefore, the routine use of this test to screen patients prior to an IVF cycle in order to select appropriate treatment (i.e., ICSI) cannot be recommended.

Effect of sperm-agglutinating antibodies on sperm capacitation and acrosome reaction.

OBJECTIVE: To study the effect of polyclonal/monospecific antisera on sperm agglutination versus capacitation as well as acrosome reaction. MATERIALS AND METHODS: Swim-up spermatozoa from cauda epididymides of fertile male hamsters were incubated under liquid paraffin with polyclonal/monospecific antisera obtained from immunized BALB/C mice, as well as with normal serum from control BALB/C mice, at various dilutions. RESULTS: The anti-sperm antibodies caused a significant (P < .05) sperm agglutination of various types of dilutions below 1:1000. Both capacitation and true acrosome reaction were inhibited significantly in the spermatozoa incubated with polyclonal/monospecific antisera. Capacitation in the spermatozoa with normal serum started earlier, i.e., at 2 hours of incubation compared to 3 hours of incubation in controls. CONCLUSION: The data differentiate the sperm agglutinating activity from anticapacitation and antiacrosome reaction activity of antisperm antisera at 1:1000 dilution.