Subfertile Men Research Articles

Office Based Screening of Sperm Autoimmunity

Sperm autoimmunity has been reported as a contributory cause of male factor infertility in up to 10% of subfertile men. The standard laboratory technique for the determination of sperm bound antisperm antibodies is the direct immunobead test. However, because of its complex methodology this test does not readily lend itself as a screening tool for this diagnosis. We describe a simple office based procedure, the sperm mixed antiglobulin reaction, for the detection of antisperm antibodies and compare its diagnostic accuracy with the standard direct immunobead test. Both assays were simultaneously performed on each of 102 semen specimens from men with male factor infertility. The rate of detection of sperm autoantibodies was 16.6% and 19.6% by the direct immunobead test and sperm mixed antiglobulin reaction, respectively. Compared to the direct immunobead test, sperm mixed antiglobulin reaction demonstrated a sensitivity and specificity of 100% and 96%, respectively. The positive predictive value was 85% and the negative predictive value was 100%, emphasizing its particularly useful role as a screening tool for office based detection of sperm autoantibodies in men who present for evaluation of fertility potential.

Subfertility in men Measurement error and confounding not accounted for

EDITOR, - Richard Lilford and colleagues conclude that subfertility in men is familial and that their observations are consistent with a genetic cause.1 Measurement error and confounding may provide alternative explanations. In case-control studies it is essential that the data collected from cases and controls should be of comparable accuracy - a requirement not met by this study, in which infertility in the brothers of cases and controls was assessed on hearsay evidence. The reasons for childlessness are more likely to be explored when two brothers are childless than when one brother is fertile and the other childless. Subfertile men may actively seek the explanation of their brother's childlessness in an attempt to explain their own subfertility. Cases and controls may interpret unexplained childlessness from their own experience as involuntary or voluntary. Additional bias may be introduced if the collection of information was not blinded. The use of reported childlessness also assumes that all such childlessness is due to male subfertility, the partner's contribution being ignored. Misclassification could easily explain these results. We agree that the difference in …

Effects of the coculture with human endometrial cells on the function of spermatozoa from subfertile men

Effects of the coculture with human endometrial cells on the function of spermatozoa from subfertile men

Methodological aspects of sperm morphology evaluation: comparison between strict and liberal criteria

To compare the methodological accuracy of different sperm morphology criteria. A multicenter study including 10 laboratories with high expertise in semen analysis. Semen preparations of subfertile men with a variable degree of teratozoospermia and of fertile semen donors. Detailed assessment of sperm morphology on 10 air-dried semen smears, of which 3 originated from the same ejaculate. The average coefficient of variation calculated on the three smears of the same ejaculate was higher when strict criteria for morphological normality were used and borderline cells were classified as abnormal than when criteria of abnormality were applied and borderline cells were considered normal. The correspondence between individual centers mutually and of each center with the average result of all centers was better when the latter approach was taken. The performance of two computer-assisted systems was intermediate between that of the two approaches, whereas one system gave unreliable results. Strict criteria for normality of sperm morphology, with borderline cells considered abnormal, gives results that are less reproducible and less accurate than the approach that classifies sperm as abnormal, with borderline cells considered normal.

Andrology: Morphometric characteristics of motile spermatozoa in subfertile men with an excess of non-sperm cells in the ejaculate

A comparison has been made between the morphological dimensions of motile spermatozoa in subfertile men demonstrating a persistent excess of non-sperm cells in their semen and those found in spermatozoa from fertile controls. Scanning electron microscopy and image analysis were used to make a morphometric assessment of defined sperm parameters in the study and control subjects. Half of the study cohort had an excess (> 5 x 10(6)/ml) of seminal leukocytes and the remainder an excess of sperm precursors in the ejaculate. In subjects with a sperm precursor excess, the motile spermatozoa had several significantly larger head parameters than those from both the fertile controls and those men with a leukocyte excess. Mid-piece and tail measurements did not differ significantly between groups. These findings suggest that, where there are large numbers of immature germinal elements present in semen, there is aberrant morphological development of the motile, apparently mature, spermatozoa which may represent disordered spermatogenesis.

NONSURGICAL TREATMENT OF MALE INFERTILITY: An Update

As has been shown, when well-defined pathophysiology such as hypogonadotropic hypogonadism exists in subfertile men, reasonably successful treatment options are available. When subfertility is unexplained or idiopathic, treatment is less successful. At present, numerous conditions of varying pathophysiology are lumped together as "idiopathic subfertility" and treated with the same nonspecific therapeutic modalities with minimal success. As further research elucidates the biochemical abnormalities causing these conditions, rational and specific therapies may be developed. In addition, advances in assisted reproductive technologies will allow treatment for many patients for whom no therapy currently exists.

Relationship of Sperm Parameters with Levels of Reactive Oxygen Species in Semen Specimens

The levels of reactive oxygen species were compared in semen specimens from suspected subfertile men and from normal volunteers, and correlated with other semen parameters. Reactive oxygen species formation was measured in semen samples that had no white blood cells by a chemiluminescence assay with a Luminometer. The relationship of seminal reactive oxygen species to several sperm parameters was evaluated. A total of 84 specimens from 69 suspected subfertile men and 15 normal volunteers was tested for reactive oxygen species production. Comparison of reactive oxygen species levels in white blood cell-negative patient and donor specimens showed significantly higher values (p <0.005) in the patient group. Similarly, levels in patient and donor specimens with normal sperm motility were significantly lower (p <0.005) than those in specimens showing poor motility. The seminal reactive oxygen species levels of white blood cell-negative patients with abnormal morphology were significantly higher (p <0.005) than those in white blood cell-negative patients with normal morphology. Our results show that seminal reactive oxygen species levels in suspected subfertile men are significantly higher than in normal men, and that the presence of excess reactive oxygen species in semen is positively correlated with low sperm concentration, poor motility and poor morphology. In conclusion, the evaluation of reactive oxygen species levels in cases of idiopathic male infertility could serve as an important marker of sperm dysfunction.

Unilateral testicular obstruction: orchidectomy or reconstruction?

Unilateral testicular obstruction can cause infertility in man by reducing sperm output, or by stimulating antisperm antibody production. Amongst 125 subfertile men with unilateral blocks, the causes were defined as post-infective (n = 72), post-traumatic (n = 41) or congenital (n = 12) by surgical exploration. Of these, 50 had severe oligozoospermia or azoospermia, and another 30 had oligozoospermia; 67 had significantly raised serum antisperm antibody titres. The outflow tracts were reconstructed in 87 men, while in 24 the obstructed testis was removed and replaced with a prosthesis. Follow-up information is available in 104 men. Reconstruction produced significant improvement in sperm concentration in the ejaculate but little change in antisperm antibody titres. Orchidectomy lowered seminal plasma antibody titres and had no effect on sperm output. Orchidectomy plus prednisolone was most effective in terms of pregnancies produced in the female partners. Testicular biopsies showed that spermatogenesis was normal on the side of the block in 90% of cases, whereas it was impaired in the contralateral testis in 40% of men. Although sperm output can be improved by reconstruction, the stimulus to antibody production is more reliably eliminated by orchidectomy. Sperm concentration, antisperm antibody titres and bilateral testicular biopsy results should be taken into account before making a final therapeutic recommendation in men with unilateral testicular obstruction.

Pharmacological stimulation of sperm motility

The treatment of male factor infertility is a rapidly developing field. The introduction of microsurgical fertilization techniques allows assisted conception units to treat couples who previously would not have benefited from in-vitro fertilization techniques. However, these techniques are only used for the minority of subfertile men in andrological practice. Many subfertile men are still treated pharmacologically or by sperm selection methods to enhance sperm fertilizing ability. Numerous pharmacological compounds have been described that enhance sperm motility and thus, potentially, sperm fertilizing capacity. This paper attempts to review these compounds and assess their role in treatment of the subfertile male.

Incidence of Antisperm Antibodies in Patients with Carcinoma of the Testis and in Subfertile Men with Normogonadotropic Oligoasthenoteratozoospermia

The incidence and the clinical relevance of sperm-reactive antibodies in subfertile men and in testicular cancer patients were assessed in a pilot study. The sera of 42 men with normogonadotropic oligoasthenoteratozoospermia (OAT syndrome, n = 20) or carcinoma of the testis after inguinal semicastration (n = 22) were analyzed for agglutinating antisperm antibodies using fluorescein-labeled antiglobulin. In the group with the OAT syndrome, the incidence of sperm-reactive antibodies was only 5%, which is comparable to that in normal fertile men. Although the incidence of 18% in the testicular cancer patients was markedly higher, only 2 of the patients in question had abnormal spermiograms, which in one case could, moreover, be explained by previous radiation therapy. In summary in this small group of patients, serum monitoring for sperm-reactive antibodies appeared to be of limited clinical relevance in patients with the OAT syndrome and in testicular cancer patients.

Effects of the coculture with human endometrial cells on the function of spermatozoa from subfertile men

To evaluate the effects of a coculture with human endometrial cells on the function of spermatozoa from samples obtained from infertile couples. In a prospective study, human spermatozoa selected by swim-up from fresh samples were cultured on human endometrial feeder layers. Thereafter, their viability, motility, acrosome integrity, and ability to penetrate zona-free hamster oocytes were evaluated. Spermatozoa from the same samples incubated under the same conditions but in the absence of endometrial cells, as well as in the medium previously spent for cell culture, were used as controls. Andrology Laboratory of the Infertility Center of San Raffaele Hospital. Spermatozoa were obtained from 17 infertile men attending the Infertility Center at our hospital. Spermatozoa incubated in the presence of endometrial cell feeder layers did not differ from controls with regard to their viability or motility. Conversely, the percent spontaneous acrosome reactions after 18 hours of incubation was significantly higher for spermatozoa cocultured (19.7 +/- 2.2 versus 11.2 +/- 1.9; mean +/- SE). The mean number of spermatozoa penetrating hamster oocytes was also significantly improved (1.24 +/- 0.3 versus 0.68 +/- 0.24). This effect did not seem to be solely due to the secretion of soluble factors by endometrial cells in the medium, in that spermatozoa incubated in the medium spent for endometrial cell culture had a similar acrosome reaction percentage but a lower rate of hamster egg penetration. The coculture with human endometrial cells appeared to be beneficial for improving the sperm function. This effect partially may be due to the secretion of steroids in the medium, which increases the quota of spontaneous acrosome reaction and in part due to the direct contact of cells with spermatozoa, maybe for the detoxification of the medium or the release of trophic factors. Coculture might be a promising approach to preparing spermatozoa for assisted fertilization in cases of subfertile males.

Effect of low-dose prednisolone on sperm fertilizing capacity in subfertile men with circulating antisperm antibodies.

To assess if low-dose prednisolone reduced circulating antisperm antibodies and improved sperm fertilizing capacity, the hamster oocyte penetration test (HOPT) was used to evaluate treatment in 36 males with significant serum antisperm antibodies, measured by the tray agglutination test (TAT). Prednisolone 5 mg three times a day for 3 months was used. Only couples in whom all tests in the female partner were normal were entered into the study. A significant increase in sperm density, normal morphology, and HOPT were noted after therapy (p < .001). A significant decrease in antisperm antibody titer was noted (p < .0001) and correlated with improvement of HOPT (p < .05). There were no significant side effects. Six pregnancies (17%) occurred. Three pregnancies (18%) occurred in partners of an untreated group of 17 men. Prednisolone therapy in this regime does not significantly improve pregnancy rates. The HOPT does not offer any additional information for predicting patients who will show an improvement in antibody titers or achieve pregnancy after steroids.

Improvement in membrane integrity and acrosin levels of human sperm by use of L4 membrane.

Previous results have demonstrated the advantage of using an L4 filter as a method for sperm preparation is assisted reproductive techniques. The present study examined the effect of two sperm processing methods (L4 filter and swim-up) on sperm acrosin levels and the integrity of the sperm membrane as measured by the hypoosmotic swelling (HOS) test. Semen specimens from 35 men (19 suspected subfertile men and 16 normal volunteers) were used to measure acrosin activity; 30 specimens (14 patients and 16 normal men) were used to assess the integrity of sperm membrane by the HOS test. The acrosin activity of L4-filtered spermatozoa (mean 110 +/- 85 microIU acrosin/10(6) sperm) in patients and donors was significantly higher (p < .002) than in either the swim-up specimens (76 +/- 58 microIU) or original unprocessed ejaculate (46 +/- 27 microIU). Similarly, the percentage of tail swelling measured by HOS test was significantly higher (p < .01) in the L4-filtered sperm (mean 71 +/- 13%) than in either swim-up specimens (59 +/- 18%) or original ejaculate (61 +/- 10%). These results clearly demonstrate improvement in two important functional parameters of sperm fertilizing ability in sperm processed by the L4-filter technique, thereby providing new evidence in support of the use of L4 filters for sperm processing.

Was Young's syndrome caused by exposure to mercury in childhood?

To determine whether the incidence of chronic sinusitis, bronchitis, or bronchiectasis in men with obstructive azoospermia (Young's syndrome) has fallen in men born after 1955 when calomel (mercurous chloride) was removed from teething powders and worm medication in the United Kingdom. A prospective study of aetiological factors in subfertile men with epididymal obstruction operated on between 1975 and 1993. Central London. 274 men with obstructive azoospermia undergoing epididymovasostomy; date of birth was recorded and illness in childhood, persistent nasal or respiratory symptoms, and previous urinary or genital infection were asked about. Site of epididymal block and association with possible aetiological factors, related to date of birth. 146 men had hold up in the head of the epididymis (capital blocks): 119 (82%) had Young's syndrome, and 11 gave a definite history of pink disease (mercury intoxication) in childhood. 128 had obstruction lower down towards the tail of the epididymis (caudal blocks): 64 (50%) had a history of genital or urinary infection, and only three had Young's syndrome; none had had pink disease. The incidence of Young's syndrome fell significantly from 114 (50%) of 227 men born up to 1955 to eight (17%) of 47 men born since then. The decline in incidence of Young's syndrome in those born after 1955 is similar to that observed with pink disease, suggesting that both conditions may have had a similar aetiology--mercury intoxication.

The effects on in-vitro fertilization of autoantibodies to spermatozoa in subfertile men

Thirty-six infertile couples underwent treatment by in-vitro fertilization. In 16 couples (group 1) the male partner was positive for antisperm antibodies measured by direct mixed antiglobulin reaction, direct immunobead test, and serum and/or seminal plasma tray agglutination test. In 20 couples (group 2) the men had no such antibodies. Men with poor sperm motility were excluded. The female partners had no antisperm antibodies, and in the controls (group 2) infertility was due to a known female factor. The fertilization rate in couples without antisperm antibodies (group 2) was 72.7% compared to 50.5% when the men had antibodies. However, the pregnancy rate per embryo transfer was not significantly different in the two groups (46.1% in group 1, 33.3% in group 2). This indicates that antisperm antibodies in the male interfere with sperm--egg fusion and subsequent fertilization but once fertilization has occurred, the pregnancy rate remains the same.

Novel parameters of human sperm hypoosmotic swelling test and their correlation to standard spermatogram, total motile sperm fraction, and sperm penetration assay

To improve the accuracy of human sperm hypoosmotic swelling test by introducing new morphological evaluation parameters for this test. Individual semen samples were processed, and the standard spermatogram, total motile sperm fraction, sperm penetration assay (SPA), and hypoosmotic swelling test were performed. Male infertility clinic and andrologic laboratory in a university medical center. One hundred eighteen subfertile men undergoing an infertility work-up. The results of hypoosmotic swelling test were subdivided into four groups (hypoosmotic swelling test 1, 2, 3, 4) according to the different shapes of hypoosmotically affected spermatozoa tails. The hypoosmotic swelling test 3 had the best correlation with the different sperm parameters: motility, total motile sperm fraction, concentration, and SPA. The hypoosmotic swelling test 1 showed a satisfactory interrelation for concentration and for total motile sperm fraction but no correlation for motility and SPA. The hypoosmotic swelling test 2 and the hypoosmotic swelling test 4 demonstrated no correlation at all with motility, concentration, morphology and SPA. The precision of hypoosmotic swelling test may be improved by using the hypoosmotic swelling test 3 type of spermatozoa tail shape as the parameter for a normally functioning spermatozoa.

Variations of testicles of subfertile men with epidemic patiditis in anamnesis

The investigation data of testicles bioptates of 153 subfertile men are given. The examined patients are divided into 2 groups: the first group includes subfertile men with epidemic parotidits in anamnesis (91), the second one includes subfertile men, never being ill with epidemic parotiditis (62). The peculiarity lies in the fact that the morphologic variations of testicles of subfertile men with epidemic parotiditis in anamnesis are similar to the variations of autoimmune nature.

Differentiation of round cells in semen by means of monoclonal antibodies and relationship with male fertility

To differentiate round cells in semen samples of subfertile men and evaluate the clinical significance during infertility investigation. One hundred and eight randomly chosen couples with a median duration of infertility of 4 (range, 1 to 20) years presenting at the outpatient infertility clinic of the University of Heidelberg, Germany. Differentiation of round cells in semen by means of monoclonal antibodies (mABs) and a streptavidin-biotin system for staining. Correlation of results with medical history, outcome of clinical examination, sperm analysis, microbial screening of both partners, evaluation of sperm functional capacity in vivo by means of the postcoital test (PCT) and in vitro with the standardized crossed sperm-cervical mucus penetration test (SCMPT) and the subsequent fertility in a prospective study. The method used for differentiation of round cells proved to be practical and suitable for routine use. The percentage of leukocytes ranged from 0% to 58% with a median of 3%. Number of round cells and percentage of leukocytes did not differ markedly with regard to andrologic history, clinical findings, for example, varicocele, results of standard sperm analysis, and microbial colonization of semen samples. However, high rates of leukocytes of the round cells correlated with reduced sperm count and morphology and results of PCT. Leukocyte-positive (> 15% leukocytes) specimens were also significantly more frequent in case of inadequate SCMPT and reduced sperm penetration ability in vitro. In asymptomatic patients (in terms of genital tract infection), the majority of round cells consist of immature germ cells and < 5% are white blood cells. The streptavidin-biotin system and the mABs used in this study proved to be useful to identify patients with elevated rates of leukocytes in semen possibly reflecting subclinical genital tract infection with influence on sperm functional capacity and subsequent fertility. Thus the procedure can be recommended to be included in a comprehensive evaluation of male fertility.

Detection and quantitation of sperm-bound antibodies by flow cytometry of human semen.

Detection of sperm surface antibodies is important for the diagnosis and treatment of infertility. The authors have investigated the methodologic aspects of flow cytometry (FCM) to detect sperm-bound antibodies and to quantitate the sperm antibody load (antibody molecules/spermatozoa). To obtain reliable results, dead spermatozoa must be excluded from analysis because they can bind antibody nonspecifically, and comprise 10% to 58% (n = 28) of the ejaculate in subfertile men. Flow cytometry estimation of dead cells correlates (r = 0.83) significantly with the manual Eosin Y method. After staining washed sperm samples (n = 26) with fluorescein-isothiocyanate-conjugated F(ab')2 fragments of anti-IgG and IgA antibodies, the sperm load was 11,500 +/- 8,600 IgG molecules and 13,200 +/- 9,500 IgA molecules per spermatozoa. The sperm antibody load measured on different occasions could be compared between patients or in the same patient by the use of calibration standards. Since the inter- and intra-assay variation of the FCM assays was less than 10%, FCM has the potential reliably and objectively to monitor the sperm antibody load during corticosteroid treatment.

Use of theophylline to enhance sperm function.

Poor sperm motility is an important factor in male infertility. Preliminary results in our laboratory on a group of 19 men (10 suspected infertile men and 9 fertile donors) showed stimulation of sperm fertilizing ability after sperm washing with theophylline as demonstrated by zona free hamster egg penetration test. The egg penetration rate for the control spermatozoa samples from subfertile men was 16%. Incubation with theophylline (10 mM) increased the penetration rate to 46%, whereas semen incubation with theophylline (20 mM) increased the penetration rate to 51%. A similar twofold increase in egg penetration was observed in the semen of fertile men incubated with theophylline of similar concentrations. Subfertile patients with ejaculate volumes of less than or equal to 1 ml or total motile sperm count of less than or equal to 10 x 10(6)/mL or increased semen viscosity did not exhibit beneficial effects with theophylline washing as measured by hamster egg penetration test score. The increase in percentage of penetrated eggs with theophylline use in both fertile and subfertile men was significant at 10 mM concentration (p less than .001) and 20 mM (p less than .001) when compared to control (untreated) samples. No significant difference in penetration rate was seen between 10 and 20 mM theophylline concentrations. It appears that theophylline may be useful in improving the fertilizing capacity of selected human semen samples with poor motility and poor penetration ability under artificial insemination conditions.