White Blood Cells In Semen Research Articles

Male infertility: Decreased levels of selenium, zinc and antioxidants

In this study, we aimed to compare the level of zinc, selenium, glutathione peroxidase activity and antioxidant status in following populations of men: severe inflammation in prostate (>106 white blood cells in prostate secretion; n=29), severe leukocytospermia, (>106 white blood cells in semen; n=31), mild inflammation, (0.2–1M white blood cells in semen or prostate secretion; n=24), non-inflammatory oligozoospermia (n=32) and healthy controls (n=27). Male partners of infertile couples had reduced level of antioxidative activity, selenium and zinc in their seminal plasma. Most importantly, reduced selenium levels were evident in all patient groups regardless of inflammation status. Therefore, these patients might gain some benefit from selenium supplementation.

Targeting Trojan Horse leukocytes for HIV prevention

…. is sure design'd, by fraud or force: trust not their presents, nor admit the horse. Virgil, Aeneid Human immunodeficiency virus type 1 (HIV-1), the lymphotropic virus that causes AIDS, has infected more than 60 million people worldwide since its clinical appearance in 1981. Despite intensive prevention efforts, the HIV/AIDS epidemic continues to spread, particularly in developing countries in sub-Saharan Africa, southeast Asia and the Caribbean, as well as the developed world [1]. Although HIV can be transmitted very efficiently parenterally, the advent of routine blood screening prior to transfusion and harm reduction programs for injection drug users, have made this mode of transmission much less common than mucosal transmission. Most new HIV infections are attributable to mucosal transmission: through genital and rectal mucosae in the case of sexual transmission and through oral or gastrointestinal mucosae in the case of mother-to-child transmission [2]. Much has been learned about HIV pathogenesis and infection mechanisms at the molecular level, but the scientific community has yet to develop an effective vaccine or microbicide for HIV prevention. Many unanswered questions remain concerning HIV-1 sexual transmission. In 1983, barely 2 years into the AIDS epidemic, we hypothesized that the agent that was subsequently identified as HIV-1 may be sexually transmitted by infected ‘Trojan Horse’ leukocytes in semen [3]. This hypothesis was based on our knowledge at the time that human semen contains substantial numbers of T lymphocytes and macrophages, which could host a T-cell tropic virus, and the following assumptions: intracellular virus would be better protected than free virus from adverse effects of antiviral factors in the genital environment such as antiviral antibodies likely to be present in genital secretions of the virus-infected transmitter, as well as antimicrobial peptides that play an important role in genital innate immune defense; and virus-infected allogeneic cells could also escape early detection by major histocompatibility complex (MHC)-restricted cytotoxic T cells in a new host. Over the intervening 25+ years, others have also championed this cause [4,5], and convincing evidence has emerged from clinical research as well as in-vitro and animal studies that infected leukocytes indeed play a role in HIV transmission. Yet, most recent research on sexual HIV transmission has focused on cell-free HIV in genital secretions because of the wide availability of HIV RNA quantification assays. Furthermore, the majority of HIV vaccines and microbicides have been designed to block transmission of cell-free virus and have been tested in animal and in-vitro models that use cell-free virus as the only infectious inoculum. As the molecular events underlying cell-associated HIV transmission differ from those underlying cell-free virus transmission, many of the current vaccine and microbicide candidates might not be expected to protect against cell-associated HIV transmission. The failure of several recent vaccine and microbicide clinical trials may be due in part to this oversight. It should be possible to design strategies that block cell-associated HIV transmission as well as cell-free HIV transmission. In this article, we present an overview of research that has been conducted on cell-associated HIV mucosal transmission and recommendations for future research. We focus on sexual HIV transmission, but this review also has relevance for mother-to-child HIV transmission, which may occur through mucosal transmission of cell-associated HIV from maternal genital or mammary gland secretions [6–8]. We review published reports that describe and enumerate HIV-infected cells in genital secretions, and compelling evidence from clinical, animal and in-vitro studies demonstrating that such cells can transmit HIV across genital tract epithelial surfaces; potential molecular mechanisms underlying cell-associated HIV transmission that could be specifically targeted by future HIV prevention strategies; and in-vitro and animal cell-associated HIV transmission models currently used for studies on cell-associated HIV transmission mechanisms and for testing vaccine and microbicide candidates. Using this information as a foundation, we discuss the evidence and probability that various current microbicide and vaccine approaches prevent cell-associated HIV transmission, and suggest additional microbicide and vaccine concepts and experiments that will move this field forward.

Leukocytospermia in male infertility patients in China.

Recent studies have revealed a high prevalence of leukocytospermia (> 1 x 10(6) white blood cells ml-1 semen) in male infertility patients in the USA and certain European countries, and have implicated white blood cells as a cause of infertility. Since leukocytospermia may often be attributed to male genital-tract infections, its prevalence could vary widely in different populations depending on factors such as sexual practices and the prevalence of sexually transmitted pathogens. In the study described here the incidence of leukocytospermia was determined in a group of 101 male infertility patients and a small reference group of normal fertile men in Beijing, China. Seminal white blood cells (WBC) and WBC sub-populations were enumerated by peroxidase staining and immunohistological assay. Eight out of 101 (7.9%) samples from infertility patients and 0/10 samples from fertile donors were leukocytospermic. The incidence of leukocytospermia in the Chinese infertility patients was considerably lower than the 23% incidence observed in a recent study of infertility patients in the USA using a similar technique. All but one of the patients with leukocytospermia had a poor sperm count and/or poor sperm motility. However, due to the low incidence of leukocytospermia and the small number of patients in this group, a statistically significant association between leukocytospermia and poor semen quality was not attained. The simple peroxidase test correlated well with the more expensive and technically demanding immunohistological assay for detection of white blood cells in semen.

Ketotifen improves sperm motility and sperm morphology in male patients with leukocytospermia and unexplained infertility

In an open, uncontrolled study, the effect of 12 weeks of daily administration of ketotifen, an antihistamine-like drug with a mast cell stabilizing effect, on the semen quality of 55 men with leukocytospermia and unexplained infertility was examined. After 4 weeks of treatment, white blood cell count dramatically diminished and was accompanied by a significant improvement in sperm motility. A significant increase of morphologically normal sperm cells was observed at 8 weeks of treatment, and these changes remained until at least 4 weeks after the end of treatment.

Human immunodeficiency virus type-1 episomal cDNA in semen

BackgroundEpisomal 2-long terminal repeat (LTR) HIV-1 cDNA, a by-product of HIV-1 infection, is used in clinical trials as a marker for ongoing viral replication. It would be useful to employ 2-LTR cDNA to monitor cryptic HIV-1 infection in the genital tract of men on antiretroviral therapy (ART) to predict the evolution of sexually transmissible drug-resistant HIV-1, but studies thus far have failed to detect this marker in semen. The objectives of this study were: 1) to use a technique that maximizes DNA recovery from HIV-1 infected white blood cells in semen to determine if episomal 2-LTR cDNA is detectable in semen of ART-naïve men with other evidence of genital tract HIV-1 infection, and 2) to compare levels of HIV-1 2-LTR cDNA, RNA, and proviral DNA in semen from HIV-1+ men on ART.ResultsUsing a somatic cell DNA extraction technique, 2-LTR cDNA was detected by PCR/ELISA in 4 out of 8 semen samples from ART-naïve men selected for other signs of seminal HIV-1 infection (positive controls). Southern blot and DNA sequencing confirmed that the amplified sequences were HIV-1 2-LTR cDNA; copy numbers ranged from 55 to 504 copies/sample. Two semen samples from a cohort of 22 HIV-1-infected men on dual nucleoside therapy, one with and one without detectable seminal HIV-1 RNA, were 2-LTR cDNA positive (336 and 8,560 copies/sample). Following addition of indinavir to the therapy regimen, no semen samples from 21 men with controlled peripheral and seminal viral loads were 2-LTR cDNA positive at 1 and 6 month time points, despite the persistence of HIV-1 proviral DNA+ semen cells and seminal cytomegalovirus (CMV) shedding in some cases. However, one individual who failed indinavir therapy and later developed distinct protease inhibitor (PI) drug resistance mutations in semen, maintained elevated levels of HIV-1 RNA and 2-LTR cDNA in semen.Conclusion2-LTR HIV-1 cDNA is detectable in semen of HIV-1-infected men. Two men on ART had 2-LTR HIV-1 cDNA in semen, suggesting that this marker may prove to be useful to monitor HIV-1 infection in the genital tract of men on ART to predict the evolution of drug resistance mutations in semen.

Immune-endocrine interactions and Leydig cell function: the role of cytokines.

Inflammatory disease is known to affect male reproductive function and fertility. Male accessory gland infections (MAGI) account for almost 15% of all cases of male infertility seen in infertility clinics. Infections of the male accessory glands are associated with increased counts of white blood cells in semen and elevated levels of pro-inflammatory cytokines in the semen and the testis. Numerous studies have underscored the importance of cytokines in the regulation of testicular and glandular function during pathophysiological events as well as under normal physiological conditions when cytokines act as growth and differentiation factors. The purpose of this paper is to particularly review the role of cytokines in the regulation of Leydig cell function in the testis primarily under pathophysiological conditions, and also considers clinical investigations that help to improve the evaluation and treatment of male infertility.

The effects of combined conventional treatment, oral antioxidants and essential fatty acids on sperm biology in subfertile men

We evaluated the effects of combined conventional treatment, oral antioxidants (N-acetyl-cysteine or vitamins A plus E) and essential fatty acids (FA) on sperm biology in an open prospective study including 27 infertile men. The evaluation included sperm characteristics, seminal reactive oxygen species (ROS), FA of sperm membrane phospholipids, sperm oxidized DNA (8-OH-dG), and induced acrosome reaction (AR). Treatment did not improve sperm motility and morphology, nor decrease the concentration of round cells and white blood cells in semen. Sperm concentration increased in oligozoospermic men (7.4±1.3 to 12.5±1.9 million/ml). Treatment significantily reduced ROS (mean±SEM) (775.3±372.2 to 150.3±105.2×103counts/10 second) and 8-OH-dG (45.3±10.4 to 16.8±3.3 fmol/μg DNA). Treatment increased the AR (55.1±2.2 to 71.6±2.2%), the proportion of polyunsaturated FA of the phospholipids, and sperm membrane fluidity. The overall pregnancy rate was 4.5% in 134 months. The per month pregnancy rate tended to be higher in partners of (ex)-smokers (7.15%, n=14,70 months) than in never-smokers (1.6%, n=13,64 months) (OR:4.57, 95% Cl:0.55–38.1).

Role of cytokines in testicular function.

Inflammatory disease has been established to affect male reproductive function and fertility. Relevant inflammatory diseases include general and chronic infectious diseases as well as localized acute or chronic infections of the male genitourinary tract. Male accessory gland infections account for almost 15% of all cases of male infertility seen in infertility clinics while fertility usually is not a clinical objective among patients with acute systemic infections such as Gram-negative sepsis. Infections of the male accessory glands frequently are associated with increased counts of white blood cells in semen and elevated levels of proinflammatory cytokines in semen and the testis. There is a mounting body of evidence that demonstrates the importance of cytokines and chemokines in the regulation of testicular and glandular function during pathophysiological states as well as under normal physiological conditions when cytokines act as growth and differentiation factors. The purpose of this review is to examine the role of cytokines in the regulation of steroidogenesis and spermatogenesis in the testis under physiological and pathophysiological conditions and considers clinical investigations that help to improve the evaluation and treatment of male infertility.

Immature spermatids are not prevalent in semen from men who are receiving androgen-based contraceptive regimens

Objective: To determine whether immature spermatids increase in semen in response to hormonal contraceptive treatments. Such a finding would support the existence of a defect in spermiogenesis, which in turn may explain the reported variability in sperm output. Design: Semen smears were obtained from healthy men undergoing randomized control trials of T plus progestin contraceptive treatments. Patient(s): Healthy men (21–49 years) with normal semen analyses received T (50–100 mg IM weekly) in combination with either desogestrel (150–300 μg daily, ( n = 5) or levonorgestrel (125–250 μg daily, n = 10) for 24 weeks. Semen smears were made during spermatogenic suppression and recovery. Nine control subjects were also assessed. Main Outcome Measure(s): Semen analyses were performed using World Health Organization criteria. Immature spermatids and white blood cells in semen were identified by immunostaining with monoclonal antibodies to the human intra-acrosomal antigen SP-10 and the ubiquitous white cell CD-45 antigen, respectively. Result(s): In a total of 14 normal ejaculates (9 control and 5 pretreatment) 74 ±14 million/mL sperm (mean ±SEM) were seen together with a few immature spermatids (0.69 ± 0.20 million/mL). During contraceptive treatments, spermatid number decreased in parallel with the sperm concentration and spermatids disappeared in most subjects. No significant changes were seen in either leukocyte or immunonegative round cell concentration (0.41 ± 0.25 and 0.25 ± 0.09 million/mL in controls, respectively) in response to treatments. Conclusion(s): Spermatid sloughing, as assessed by the ejaculation of immature spermatids, is not a feature of T-induced spermatogenic regression in men; rather, the decline in both mature and immature germ cells in the ejaculate probably results from a decline in the number of precursor cells, ultimately resulting in severe oligoor azoospermia. Detailed studies on the sites of spermatogenic interruption are required to understand the variability in responses seen after contraceptive therapies in men.

Evaluation of beta-endorphin and interleukin-6 in seminal plasma of patients with certain andrological diseases.

Human semen contains large amounts of opioid peptides and cytokines. We have measured the concentrations of interleukin (IL)-6 in 140 semen samples and of beta-endorphin in 77 semen samples. The median concentration of beta-endorphin in seminal plasma from normozoospermic men (n = 23) was 154.7 pg/ml (10th-90th percentiles, 42.0-774.6), and there was no significant difference in the beta-endorphin concentration among normozoospermic, oligozoospermic (n = 28), asthenozoospermic (n = 15), azoospermic (n = 4) and post-vasectomy (n = 7) samples. There was no correlation between beta-endorphin concentration and sperm characteristics, nor with blood hormones. beta-Endorphin concentration was lower in cases with immunological infertility, as revealed by a positive direct mixed antiglobulin reaction test (n = 12) (P < 0.01), than in matched controls. The median concentration of IL-6 in samples with normal sperm concentration, motility and morphology with or without white blood cells (n = 39) was 26.1 pg/ml (10th-90th percentiles, 7.3-172.3), and there was no significant difference in the IL-6 concentration among normozoospermic, oligozoospermic (n = 46), asthenozoospermic (n = 32), azoospermic (n = 13) and post-vasectomy (n = 10) samples. The IL-6 concentration was significantly higher in cases of varicocele (n = 22) without white blood cells in semen (P < 0.001) than in matched controls without varicocele (n = 23). In addition, the IL-6 concentration was elevated (P < 0.0001) in cases with accessory sex gland inflammation (n = 40). IL-6 concentration was positively correlated with white blood cells in semen (n = 60, r = 0.59, P < 0.0001), but there was no correlation with beta-endorphin concentration. The IL-6 concentration chosen to differentiate between cases with and without accessory gland inflammation was 45.3 pg/ml, with a specificity of 80.6% and a sensitivity of 92.5%. It is concluded that beta-endorphin in seminal plasma plays an immune suppressive role, and that increased IL-6 concentration may be related to testicular dysfunction in cases with varicocele. Furthermore, IL-6 is an accurate marker of accessory sex gland inflammation.

The biologic significance of white blood cells in semen

To analyze the data available on the biologic significance of white blood cells (WBC) in semen of infertility patients. The relevant literature was reviewed. It is not possible to identify reliably WBC by conventional sperm staining techniques. The peroxidase method is sufficient for quantification of granulocytes, but immunocytology is the gold standard for the detection of all WBC populations in semen. Granulocytes are the most prevalent WBC type in semen (50% to 60%), followed by macrophages (20% to 30%) and T-lymphocytes (2% to 5%). The prevalence of leukocytospermia (> 10(6) WBC/mL semen) among male infertility patients is approximately 10% to 20%. There is controversy on the significance of WBC in semen. Whereas some authors did not observe sperm damage in the presence of leukocytospermia, others have found evidence that WBC are significant cofactors of male infertility: [1] seminal WBC numbers were higher in infertility patients than among fertile men; [2] leukocytospermia was associated with decreased sperm numbers and impaired sperm motility; [3] WBC damaged sperm function and hamster ovum penetration in vitro and were important prognostic factors for IVF-ET failure. Because of absence of clinical symptoms, the origin of WBC is difficult to determine. Normally, most WBC appear to originate from the epididymis because vasectomized men show very few WBC in semen. On the other hand, leukocytospermic samples show low citric acid levels, pointing to asymptomatic prostatitis as a source of WBC in semen. Surprisingly, approximately 80% of leukocytospermic samples are microbiologically negative. In some cases Chlamydia trachomatis might have triggered a persistent inflammatory reaction leading to leukocytospermia. Sperm damage by WBC can be mediated by reactive oxygen species, proteases and cytokines. Furthermore, genital tract inflammation facilitates the formation of sperm antibodies. As seminal plasma has strong anti-inflammatory properties and because there is only short contact between sperm and WBC in prostatitis and seminal vesiculitis, inflammations of the epididymis and testis are likely to have the largest impact on sperm. There is ample evidence that WBC can affect sperm function. Further studies are needed to define cofactors that increase or decrease the risk of sperm damage by WBC.

A method of human semen centrifugation to minimize the iatrogenic sperm injuries caused by reactive oxygen species.

Current techniques of sperm preparation for in vitro fertilization or intrauterine insemination require centrifugation of human semen to separate spermatozoa from the seminal plasma. Centrifugation increases reactive oxygen species (ROS) formation in semen. Moreover, high levels of ROS are associated with sperm membrane injury through spontaneous lipid peroxidation, which may alter sperm function. We investigated the relationship between centrifugation variables (time and g-force) and ROS production to establish an optimal centrifugation protocol for sperm preparation techniques. Semen from 38 men (24 patients and 14 normal volunteers) was evaluated for the formation of ROS before centrifugation and after centrifugation at 200 g for 2 or 10 min and after 500 g for 2 or 10 min. The absence of white blood cells in semen which can also produce ROS was determined with the myeloperoxidase technique (Endtz test). All specimens were negative (< 1 x 10(6)/ml) by the Endtz test. The formation of ROS was measured by chemiluminescence. ROS formation was regarded as high (positive) when the chemiluminescence response was at least 10 x 10(4) counted photons/min (cpm). The sperm concentration in each sample was adjusted to 15-20 x 10(6) cells/ml before analysis. Eight specimens (7 patients and 1 donor) exhibited high levels of ROS before centrifugation. All 8 showed further, significant increases in ROS formation regardless of g-force or time. The increase in ROS was significantly less when semen was centrifuged for 2 as compared to 10 min (p < 0.001). Six specimens previously ROS-negative became ROS-positive after centrifugation for 10 min at 200 and 500g. We conclude that the time of centrifugation is more important than g-force for inducing ROS formation in semen. Based on these results, we recommend a shorter centrifugation period in the preparation of sperm for assisted reproductive techniques.

White blood cells in semen affect hyperactivation but not sperm membrane integrity in the head and tail regions

The presence of high numbers of peroxidase-positive PML in ejaculated semen significantly reduced sperm HA, an important step leading to sperm capacitation. Sperm membranes at both the head and tail regions, as assessed by the hypo-osmotic viability parameter and the hypo-osmotic sperm swelling test, respectively, were not affected by peroxidase-containing leukocytes. Sperm motility was not affected, but sperm curvilinear and straight line velocity parameters were reduced in the presence of high concentrations of leukocytes in the ejaculate. The results suggested that the effect of leukocytes on sperm was through a reduction in sperm hyperactive motility but not through alterations in the sperm head and tail membranes.

Comparison of methods to enumerate white blood cells in semen

Seminal WBC counts obtained by an mAb-based immunohistologic method correlated well with seminal granulocyte counts obtained with a simple peroxidase method (rho = +0.70; P < 0.0001). However, total WBC counts were significantly higher than granulocyte counts for most samples. With the immunohistologic method, 17 of 112 samples (15.2%) contained > 10(6) WBC/mL semen, whereas the peroxidase method resulted in only 10 samples (8.9%) with > 10(6) WBC/mL. When the threshold defining leukocytospermia was set at 1 x 10(6) positive cells/mL for both methods, the specificity of the peroxidase test compared with the immunohistology technique was 100% (10/10), but the sensitivity was only 58.8% (10/17). When the threshold for leukocytospermia in the peroxidase test was lowered to 5 x 10(5) positive cells/mL semen, the sensitivity relative to the immunohistology technique increased to 94.1% (16/17), and specificity remained 100% (16/16). Likewise, good interassay sensitivity and specificity values were obtained with thresholds of 10(6) WBC/mL for the peroxidase assay and 2 x 10(6) WBC/mL for the immunohistology assay. We conclude that either peroxidase or immunohistology assays can be used to screen for leukocytospermia, but that more research is needed to establish thresholds for pathological levels of WBC in semen using these two approaches. Total round cell counts are of no value for enumerating WBC in semen.

The Correlation between Round Cells and White Blood Cells in the Semen

It is generally recognized that white blood cells in semen are detrimental to sperm function and fertility. Traditionally, pyospermia has been suspected if greater than 10 round cells per high power field or more than 1 million round cells per ml. were identified in the semen. Since immature germ cells and white blood cells appear as round cells under wet mount light microscopy, the relationship between increased numbers of round cells in semen and true pyospermia was examined.Semen analyses were performed on specimens from 627 infertility patients. The numbers of round cells per high power field and the number of round cells per ml. of semen were recorded. Specimens from 57 patients (9%) persistently demonstrated greater than 10 round cells per high power field or more than 1 million round cells per ml. and they were subjected to immunohistochemical staining against white blood cell surface antigens. Greater than 1 million white blood cells per ml. were identified in only 20 patients (35%) and only 8 samples demonstrated more than 3 million white blood cells per ml. Of the patients with greater than 1 million white blood cells per ml. 65% had motilities of less than 60% compared to 49% of the patients with less than this amount of pyospermia. This difference was not statistically significant. Round cells counted on a per milliliter basis correlated better with results of immunohistochemical staining than did round cells counted on a per high power field basis. These data demonstrate that the majority of patients with excess round cells in the semen do not have pyospermia and, therefore, empiric antibiotic therapy without further testing will likely be unproductive.

Comparison of three methods to detect white blood cells in semen: leukocyte esterase dipstick test, granulocyte elastase enzymeimmunoassay, and peroxidase cytochemistry

Comparison of three methods for the detection of WBC in semen revealed a low concordance of positive test results. Among 557 semen samples, most positives were observed with the leukocyte esterase dipstick test (n = 95; 17.1%) followed by the peroxidase test (n = 51; 9.2%). There was little overlap between positives in the esterase-dipstick and the peroxidase method (29/117; 24.8%). With only 4 of 557 samples (0.7%), the PMN-elastase ELISA showed a surprisingly low incidence of positives. Because of lack of a gold standard, none of the three methods could be identified as superior. Because of its simplicity, specificity, and cost-effectiveness, the peroxidase method appeared most suited for clinical application.

Leukocytospermia is associated with poor semen quality

Semen samples from 179 infertility patients were analyzed for type and number of white blood cells (WBC) by a combination of immunologic techniques. Forty-one (23%) had more than 10(6) WBC/mL semen (leukocytospermia). Semen parameters in patients with less than 10(6) WBC/mL were similar to those of 15 fertile donors. In contrast, the leukocytospermic group had significant reductions in total sperm number, percent sperm motility, sperm velocity, motility index, and total number of motile sperm. Patients with high concentrations of monocytes/macrophages (greater than 5 X 10(5)/mL; n = 27) had significantly reduced ejaculate volume; patients with high numbers of T lymphocytes (greater than 10(5)/mL; n = 19) had a significant reduction in sperm velocity; and patients with high levels of granulocyte elastase in semen (greater than 1,000 ng/mL; n = 26) had significant reductions in ejaculate volume, total sperm number, and total motile sperm number. There was no correlation between the presence of antisperm antibodies and leukocytospermia. Our data suggest that leukocytospermia may occur frequently in male infertility patients and show that elevated levels of WBC in semen are associated with poor semen quality. This provides further rationale for white blood cell testing in semen of male infertility patients, since antibiotic or anti-inflammatory therapy may be helpful in appropriately selected cases.

Leukocytospermia is associated with poor semen quality

Semen samples from 179 infertility patients were analyzed for type and number of white blood cells (WBC) by a combination of immunologic techniques. Forty-one (23%) had more than 10(6) WBC/mL semen (leukocytospermia). Semen parameters in patients with less than 10(6) WBC/mL were similar to those of 15 fertile donors. In contrast, the leukocytospermic group had significant reductions in total sperm number, percent sperm motility, sperm velocity, motility index, and total number of motile sperm. Patients with high concentrations of monocytes/macrophages (greater than 5 X 10(5)/mL; n = 27) had significantly reduced ejaculate volume; patients with high numbers of T lymphocytes (greater than 10(5)/mL; n = 19) had a significant reduction in sperm velocity; and patients with high levels of granulocyte elastase in semen (greater than 1,000 ng/mL; n = 26) had significant reductions in ejaculate volume, total sperm number, and total motile sperm number. There was no correlation between the presence of antisperm antibodies and leukocytospermia. Our data suggest that leukocytospermia may occur frequently in male infertility patients and show that elevated levels of WBC in semen are associated with poor semen quality. This provides further rationale for white blood cell testing in semen of male infertility patients, since antibiotic or anti-inflammatory therapy may be helpful in appropriately selected cases.

Relationships between [formula omitted][formula omitted] semen culture and various semen and serology parameters

Semen and blood samples from 154 rams from two Montana range flocks (Flock A, vaccinated for Brucella ovis ; Flock B, nonvaccinated) were evaluated to determine the relationship between Brucella ovis ( B. ovis ) semen culture results and various semen and blood parameters. All rams utilized in this study exhibited no palpable ram epididymitis lesions. Thirteen and 25.6% of the rams tested in Flocks A and B, respectively, had positive B. ovis semen cultures. Only age of ram and ram condition scores differed (P<0.05) between flocks. No flock by semen culture interactions were detected (P>0.05) for any of the parameters evaluated. Age of ram, ram condition score, and spermatozoa rate from forward movement were unrelated (P>0.05) to B. ovis culture results. Rams with positive B. ovis semen cultures had lower sperm motility (P<0.05), higher percentage of abnormal spermatozoa cells (P<0.05), higher percentage of spermatozoa head abnormalities (P<0.01), lower percentage of live-normal cells (P<0.05), higher incidence of white blood cells in semen (P<0.01) and higher complement fixation (CF) titers (P<0.01).

Effects of white blood cells on the in vitro penetration of zona-free hamster eggs by human spermatozoa.

The presence of white blood cells in semen has been associated with male infertility. Previous studies indicate that pyospermia occurs in conjunction with decreases in sperm motility, number of normal sperm forms, and penetration rates in the zona-free hamster egg sperm penetration assay. We have evaluated the relationship of seminal white blood cells and sperm function, as reflected in the zona-free hamster egg penetration assay, and have investigated the possible mode of action of the white cells. Egg penetration rates decreased when white blood cells from fertile or potentially fertile donors were added to their sperm suspensions prior to preincubation and at insemination in the in vitro assay. Zona-free hamster egg penetration assay results were also inhibited when the supernatant from white blood cells incubated in Biggers, Whitten, and Whittingham (BWW) medium overnight were introduced to sperm-oocyte suspensions at insemination. Conversely, egg penetration rates were enhanced in samples from hypofertile individuals when white blood cell concentrations in the semen or WBC/sperm ratios were reduced, either by physical removal or as a result of antibiotic therapy. The physical presence of leukocytes, and possibly, the extracellular release of lysosomal enzymes may be responsible for the inhibitory effects in vitro. Although the mechanism(s) by which white blood cells interfere with the fertilizing capacity of spermatozoa are not clear, it is quite obvious that their presence in the in vitro environment is undesirable and can mask an individual's actual fertilizing potential.