Cytokines In Seminal Plasma Research Articles

Analysis of 12 kinds of cytokines in seminal plasma by flow cytometry and their correlations with routine semen parameters

ObjectiveTo investigate the levels of 12 kinds of cytokines in seminal plasma and their correlations with routine semen parameters. MethodsThe remaining seminal plasma samples of 134 patients undergoing routine semen examination were collected for detecting cytokines. The parameters for sperm concentration, percentage of progressively motile sperm (PR), and motility were analyzed by a computer-assisted sperm analysis (CASA) system. According to the results of sperm concentration, PR and motility, 134 patients were divided into the normal routine semen parameters group, oligoasthenospermia group and azoospermia group. The levels of 12 kinds of cytokines in seminal plasma, including interleukin (IL)-1β, IL-2, IL-4, IL-5, IL-6, IL-8, IL-10, IL-12P70, IL-17, interferin (IFN)-α, IFN-γ, and tumor necrosis factor (TNF)-α, were detected by flow cytometry. Two seminal plasma samples were detected for 10 times, respectively, to calculate the coefficients of variation (CV) of each cytokine. The linear range of each cytokine was measured using the standard, and the correlation coefficient (r) was calculated. ResultsThe r2 of 12 kinds of cytokines detected by flow cytometry were all greater than 0.99. The reproducibility of 2 seminal plasma samples showed that the CVs of all cytokines were lower than 15 % except for TNF-α in sample 1 (15.15 %). Seminal plasma IL-6 levels were negatively correlated with semen volume (P < 0.01). Seminal plasma IL-5 levels were positively correlated with sperm concentration (P < 0.01). Seminal plasma IL-8 levels were negatively correlated with sperm motility (P < 0.01). Seminal plasma IL-8, IL-17 and IL-12P70 levels were negatively correlated with sperm PR (P < 0.05). In addition to the significant negative correlation between IL-5 and IL-17 (P < 0.05), there was a significant positive correlation between the majority of other cytokines. The levels of seminal plasma IL-17 and IL-12P70 in the oligoasthenospermia group and IL-1β and IL-12P70 in the azoospermia group were significantly higher than those in the normal routine semen parameters group (P ≤ 0.05), while the levels of IL-10 in the azoospermia group were significantly lower than that in the normal routine semen parameters group (P < 0.05). ConclusionThere are certain correlations between seminal plasma cytokines and routine semen parameters and strong correlations between different seminal plasma cytokines, suggesting that the imbalance between seminal plasma cytokines may affect sperm quality. However, it still needs to be further confirmed by large samples and multi-center clinical studies and related basic researches.

Immune regulatory cytokines in seminal plasma of healthy men: A scoping review and analysis of variance.

Seminal plasma cytokines are associated with fertility and reproductive health, but progressing their clinical utility is hampered by absence of reference data on concentration ranges of relevant cytokines in healthy men. We employed a systematic approach to assemble current evidence on the concentrations of immune regulatory cytokines present in seminal plasma (SP) of normozoospermic and/or fertile men and evaluated the impact of different platform methodologies for cytokine quantification. A systematic literature search was performed utilising PubMed, Web of Science and Scopus. Databases were searched from inception until 30th June 2022 inclusive, using combinations of keywords pertaining to seminal fluid and cytokines, and was restricted to human participants. Original data with values reported as concentration of specific cytokines in SP of men clearly defined as fertile or normozoospermic were extracted from studies written in English. A total of 3769 publications were initially identified, of which 118 fulfilled the eligibility criteria for inclusion. A total of 51 individual cytokines are detectable in SP of healthy men. The number of studies reporting on each cytokine range from 1 to>20. The reported concentrations for many cytokines linked with fertility status, including IL6, CXCL8/IL8, and TNFA, are highly variable between published studies. This is associated with the different immunoassay methodologies utilised and may be exacerbated by a lack of validation of assays to ensure suitability for SP assessment. Due to the large variation between studies, accurate reference ranges for healthy men cannot be determined from the published data. The concentrations of cytokines and chemokines detected in SP is inconsistent and highly variable between studies and cohorts, limiting current capacity to define reference ranges for cytokine concentrations in fertile men. The lack of standardisation in methods used to process and store SP, and variation in platforms used to evaluate cytokine abundance, are factors contributing to the observed heterogeneity. To progress the clinical utility of SP cytokine analysis will require standardisation and validation of methodologies so that reference ranges for healthy fertile men can be defined.

Relationship between seminal plasma cytokine levels and asthenozoospermia in infertile men

Background. Violation of male fertility can have various causes, one of which is a decrease in sperm motility. In the pathogenesis of asthenozoospermia, some bioactive substances, in particular cytokines, may be involved.Aim. To estimate the number of the most studied cytokines to date in the ejaculate of infertile men with normozoospermia and patients with asthenozoospermia.Materials and methods. We examined 38 infertile patients with normozoospermia and 10 men with asthenozoospermia, comparable in age. All participants in the study were performed a spermogram and an analysis to determine the amount of the cytokines in seminal plasma, such as interleukins 1β, 2, 4–8, 10, 12, 13, 17, G-CSF, GM-CSF, interferon γ, MCP-1, MIP-1β and tumor necrosis factor α. Then, using nonparametric statistics, the relationship between the level of the above cytokines and the presence of asthenozoospermia was analyzed.Results. As a result, we were able to find out that there is a relationship between the concentration of interleukin 6, interleukin 8 and MIP-1β and sperm motility.Conclusion. Further research is needed to assess whether these bioactive inflammatory mediators have potential relevance for use as targets in the treatment of male infertility.

The effects of nutritional level and body condition score on cytokines in seminal plasma of beef bulls

IntroductionHigh quality semen is essential for reproductive efficiency. Nutrition and environmental factors impact the ejaculate components, like cytokines, that are essential for pregnancy establishment. We hypothesized that differing nutritional periods and body condition scores would affect seminal plasma cytokine concentrations of bulls.MethodsMature Angus bulls (n=11) were individually housed and randomly assigned to either over-fed (n=5) or restricted (n=6) treatment pathways. Bulls were fed different volumes of a single ration creating 8 individual treatment periods. Body weight and body condition scores were taken every 14 d to manage intake volumes. Ejaculates were collected every 84 d to determine seminal plasma cytokine profiles. A complete randomized design was used to evaluate seminal plasma cytokines after each nutritional treatment. Initial cytokine concentrations and volume of the ejaculate were included as covariates.ResultsAll cytokines returned to initial concentrations following maintenance treatments at an ideal body condition score of 6. Nutritional treatments affected (P ≤ 0.05) IFN-γ, IL-8, MIP-1α, MIP-1β, TNF-α, IL-1β, and VEGF-A. However, TNF-α, IFN-γ, and MIP-1α had the greatest impact on cytokine profile.DiscussionNutritional levels and adiposity altered seminal plasma cytokine concentrations which could potentially impact the inflammatory balance of the uterus and the immune responses necessary for pregnancy establishment.

Применение комплекса природных противомикробных пептидов и цитокинов при мужском бесплодии и хроническим простатите

Chronic prostatitis (CP) has long been considered one of the common causes of male infertility. The influence of therapy for CP on spermatogenesis is well studied. However, recently, especially in the era of the new coronavirus infection (COVID-19), much attention has been paid to the effects of cytokines on the pathogenesis of the inflammatory process, and their impact on male fertility.To study the effect of a complex of natural antimicrobial peptides and cytokines (Superlymph) on ejaculate, MAR test, oxidative stress (OS), and sperm DNA fragmentation (SDF) in men with infertility and a history of CP. In some patients, the level of cytokines before and after treatment was determined.A total of 583 infertile men with a history of CP, aged 18-75 years, were screened according to the inclusion/ exclusion criteria. In group I, patients received antioxidant stimulation of spermatogenesis (triovit, trental, mexidol) in combination with a complex of natural antimicrobial peptides and cytokines (Superlymph) one suppository 25 IU at night for 20 days for 1-3 weeks and for 6-8 weeks for 2 months. In group II, antioxidant stimulation of spermatogenesis was prescribed along with suppositories for the prevention of CP for 2 months. The evaluated criteria included sperm analysis, MAR test, oxidative stress in spermatozoa (reactive oxygen species (ROS)) and SDF. The levels of seminal plasma cytokines were also measured before and after treatment in 32 patients using the Bio-Plex Pro panel for the simultaneous determination of 17 human cytokines, the Human Cytokine 17-plex Assay. Statistica-10 software was used for statistical analysis (p<0.05).Of 135 eligible patients, results were collected from 125 men (mean age 31.7 +/- 13.4 years). In group I, there was a more pronounced increase in motility by 42.3%, a decrease in the MAR-test level by 64.7%, oxidative stress in spermatozoa by 56.1%, and SDF by 25.6%, compared to group II (30.4 %, 10.5%, 45.7%, 21.9%), respectively. All these changes were significant with the exception of a decrease in the level of SDF. In group I, there was a predominant increase in the activity of anti-inflammatory cytokines (IL-4, 10, 13), a decrease in the level of pro-inflammatory cytokines ((IL-2, 12). In group II, there was a significant decrease in both pro-inflammatory (IL-2, 6, 8), and anti-inflammatory cytokines (IL-4). The remaining changes were insignificant. Among mild side effects, there were a change in the color of the sclera in 4 (3.2%), anal pain in 3 (2.4%), a decrease in libido in 2 (1.6%) patients. No differences between groups in the frequency of side effects were recorded (p>0.05).Antioxidant stimulation of spermatogenesis in infertile men and CP is an effective and safe. The addition of a complex of natural antimicrobial peptides and cytokines (Superlymph) leads to a more profound increase in sperm motility, a decrease in the MAR-test, oxidative stress in sperm cells, which potentiates the effects of therapy. The use of Superlymph is accompanied by a more pronounced positive effect on pathospermia, an increase in anti-inflammatory and a decrease in pro-inflammatory cytokines. In the contrary, in control group, where patients received antioxidant stimulation of spermatogenesis, only a decrease in the level of main cytokines was observed. The effect of a complex of natural antimicrobial peptides and cytokines (Superlymph) on the level of SDF, as well as the relationship with fertility and childbirth rates, requires further study.

Diagnostic cytokine marker of male infertility - interleukin 4.

Analysis of the study is to assess the diagnostic significance of cytokines in the sperm plasma of men of reproductive age (20 - 45 years) of two groups: of patients with chronic bacterial prostatitis, not complicated by infertility and with loss of fertility. The study of sperm plasma - the WHO standard. Determination of the level of cytokines in seminal plasma - by enzyme immunoassay («Cytokine», Russia). Two methods of mathematical statistics were used: discriminant analysis and classification trees (decision trees).The similarity of interpretations of discriminant analysis and decision tree was noted, where the main role in both cases belongs to the cytokine IL-4. The level of sperm IL-4 in combination with therapeutic monitoring can be used for the medical management of patients with chronic prostatitis in order to prevent the development of infertility and to develop methods for screening diagnostics of fertility disorders in men.

Cytokine profile of seminal plasma and effectiveness of assisted reproductive technology programs

Increasing evidence shows that seminal plasma is among the most important immunoregulatory factors in female reproductive function. We suggest that the favorable effect of the partner’s seminal plasma (SP) upon pregnancy occurence in women during the cycle of in vitro fertilization (IVF)/intracytoplasmic sperm injection (ICSI) may be provided by the balanced content of Th1/Th2-dependent seminal cytokines. Otherwise, in case of pathologic changes in SP composition, it may negatively affect the IVF efficiency. Our aim was to determine whether the levels of seminal IL-1β, IL-2, IL-4, IL-5, IL-6, IL-8, IL-10, IL-12p70, IL-18, IFNγ, TNFα, TNFβ and TGF-β1 are associated with pregnancy establishment in female patients exposed to seminal plasma during IVF/ICSI cycle.Twenty-eight female patients were exposed to seminal plasma via sexual intercourse before the day of oocyte retrieval, and also underwent intravaginal application of seminal plasma just after transvaginal puncture and oocyte retrieval. Quantitative measurement of seminal cytokines was performed by FlowCytomix™ technology. IL-1, IL-18 and TNFβ concentrations were significantly higher in non-pregnant group (p = 0.011; p = 0.030 and p = 0.008, respectively). The contents of IL-2, IL-6, IL-18 and TNFβ per ejaculate were also significantly higher in non-pregnant group (p = 0.020; р = 0.042; р = 0.030 and р = 0.004, respectively). We conclude that elevated concentrations of proinflammatory cytokines in seminal plasma, as well as their total excessive content per ejaculate may have an adverse effect on implantation and pregnancy establishment.

Identification of distinct seminal plasma cytokine profiles associated with male age and lifestyle characteristics in unexplained recurrent pregnancy loss

BackgroundSeminal plasma contains a wide range of cytokines, chemokines and growth factors. Part of these signalling molecules assist in inducing a state of active maternal immune tolerance towards the fetus. Disbalances in seminal plasma content may contribute to pregnancy loss. This study investigated cytokine expression profiles in seminal plasma of male partners of couples with unexplained recurrent pregnancy loss (RPL) and the association with clinical and lifestyle characteristics, including smoking, alcohol consumption and body mass index (BMI). MethodsIn the seminal plasma of 52 men who visited a specialised RPL clinic the levels of 25 pre-selected cytokines, chemokines and growth factors were measured by Bio-Plex assay or ELISA. Two-way hierarchical cluster analysis was performed. Identified patient clusters were compared on clinical and lifestyle characteristics. ResultsTwo distinct cytokine expression profiles in the seminal plasma were revealed by cluster analysis. Patient cluster I showed relatively higher levels of pro-inflammatory cytokines, including IL-1α, IL-1β, IL-6, IL-8, IL-12, IL-18 and TNF-α, compared to Patient cluster II. Men belonging to Patient cluster I were significantly older and had significantly more lifestyle risk factors compared to men in Patient cluster II. ConclusionCluster analysis suggested the existence of a less favourable pro-inflammatory cytokine expression profile, being present in part of men affected by RPL and associated with advanced male age and lifestyle risk factors. These findings may serve as a starting point for further research into underlying mechanisms and ultimately lead to novel diagnostic and therapeutic approaches for couples with RPL.

PSI-7 Differing planes of nutrition affecting the cytokines of bovine seminal plasma in beef cattle

Abstract Bull fertility is highly influential to profits of cow-calf production; however, bulls often experience various planes of nutrition throughout the year. These changes may influence the composition of the ejaculate and their ability to get females pregnant. We hypothesized that differing nutritional planes would affect the cytokines concentration in the seminal plasma of bulls. Mature angus bulls (n = 12) were individually housed and randomly assigned to one of two treatments: 1) over-fed (n = 6) or 2) restricted (n = 6). Bulls were fed the same ration (35% ground hay, 35% cracked corn, 20% distillers’ grain and 10% soybean meal) at differing volumes to achieve desired nutritional planes. Body weight and BCS were taken every two weeks to monitor nutritional planes. Blood and ejaculate were collected once a month to determine cytokine profiles within seminal plasma. Statistical analyses were conducted with GLIMMIX procedure of SAS with bull within treatment as a random effect and time as a repeated measure to determine if nutrition level, time and the interaction influenced seminal plasma cytokine concentration. There were no significant time or treatment×time interactions for any cytokine concentrations. Interferon-γ tended to be increased (P &amp;lt; 0.10) in restricted bulls (3.21±0.95 pg/mL) compared with over-fed bulls (0.64±0.95 pg/mL). The concentrations of IL-8 also tended to be increased (P &amp;lt; 0.10) in restricted (318.89±66.42 pg/mL) compared with over-fed bulls (158.02±66.42 pg/mL). Interleukin-1β in over-fed bulls (24.77±17.51 pg/mL) tended to be decreased (P &amp;lt; 0.10) compared with restricted bulls (65.8±17.51 pg/mL). Interleukin-10 tended to be greater (P &amp;lt; 0.10) in restricted (17.95±2.29 pg/mL) than in the over-fed bulls (12.41±2.29 pg/mL). The concentration of VEGF-A was greater (P &amp;lt; 0.05) in restricted bulls (3,551.28±224.49 pg/mL) than over-fed bulls (2,715.72±224.49 pg/mL). In conclusion, cytokine concentrations were elevated by restricted nutritional levels which could impact the ability of the ejaculate to stimulate uterine gene expression and establish pregnancy.

Measurable Cytokine Concentrations in Pig Seminal Plasma Are Modified by Semen Handling and Storage.

Sample handling and storing are critical steps for the reliable measurement of circulating biomolecules in biological fluids. This study evaluates how cytokine measurements in pig seminal plasma (SP) vary depending on semen handling and SP storage. Thirteen cytokines (GM-CSF, IFNγ, IL-1α, IL-1β, IL-1ra, IL-2, IL-4, IL-6, IL-8, IL-10, IL-12, IL-18 and TNFα) were measured using Luminex xMAP® technology in individual seminal plasma (SP) samples (n = 62) from healthy breeding boars. Three separate experiments explored the delay (2 h and 24 h) in SP collection after ejaculation (Experiment 1) and SP storage, either short-term (5 °C, −20 °C and −80 °C for 72 h, Experiment 2) or long-term (at −20 °C and −80 °C for two months, Experiment 3), before analysis. Levels in fresh SP-samples were used as baseline control values. Delays in SP harvesting of up to 24 h did not substantially impact SP cytokine measurements. Some cytokines showed instability in stored SP samples, mainly in long-term storage. Ideally, cytokines in pig SP should be measured in fresh samples harvested within 24 h after ejaculation. If storage of SP is imperative, storage conditions should be adjusted for each cytokine.

Period of Boar Ejaculate Collection Contributes to the Yearly Intra-Male Variability of Seminal Plasma Cytokines.

The concentrations of cytokines in seminal plasma (SP) fluctuate over time in healthy males, weakening their practical usefulness as diagnostic tools. This study evaluated the relevance of intra-male variability in SP cytokines and to what extent the period of the year when ejaculate is collected contributes to such variability. Thirteen cytokines (GM-CSF, IFNγ, IL-1α, IL-1β, IL-1ra, IL-2, IL-4, IL-6, IL-8, IL-10, IL-12, IL-18, and TNFα) were measured using the Luminex xMAP® technology for 180 SP samples of ejaculate collected over a year from nine healthy and fertile boars. The SP samples were grouped into two annual periods according to decreasing or increasing daylight and ambient temperature. Intra-male variability was higher than inter-male variability for all cytokines. All SP cytokines showed concentration differences between the two periods of the year, showing the highest concentration during the increasing daylength/temperature period, irrespective of the male. Similarly, some cytokines showed differences between daylength/temperature periods when focusing on their total amount in the ejaculate. No strong relationship (explaining more than 50% of the total variance) was found between annual fluctuations in SP-cytokine levels and semen parameters. In conclusion, the period of the year during which ejaculates were collected helps explain the intra-male variability of SP-cytokine levels in breeding boars.

Seminal Plasma Cytokines Are Predictive of the Outcome of Boar Sperm Preservation.

Background: Boar seminal plasma is rich in cytokines, which could influence the capability of spermatozoa to tolerate preservation.Objectives: To evaluate the involvement of boar seminal plasma cytokines in the changes experienced by boar spermatozoa during their storage, either in liquid or frozen state.Materials and Methods: In two separated experiments, semen samples from healthy and fertile boars were split in two aliquots, one centrifuged twice (1,500 ×g for 10 min) to harvest seminal plasma, whereas the other was either commercially extended (3 × 107 sperm/mL) and liquid-stored at 17°C during 144 h (n = 28, Experiment 1) or frozen-thawed using a standard 0.5 mL protocol (n = 27, Experiment 2). Sixteen cytokines were quantified using Luminex xMAP®. Sperm attributes (CASA-evaluated total and progressive motility; flow cytometry-evaluated sperm viability, production of intracellular H2O2 and and levels of lipid peroxidation in viable spermatozoa) were evaluated either at 0, 72, or 144 h of liquid storage (Experiment 1) or before freezing and at 30- and 150-min post-thawing (Experiment 2).Results: Multiple linear regression models, with Bayesian approach for variable selection, revealed that the anti-inflammatory TGF-β2, TGF-β3, IL-1Ra, and IL-4 and the pro-inflammatory IL-8 and IL-18, predicted changes in sperm motility for liquid-stored semen while the anti-inflammatory IFN-γ was included in the models predicting changes in all sperm attributes for cryopreserved semen.Conclusion: Specific boar seminal plasma cytokines would contribute to modulate the structural and metabolic changes shown by spermatozoa during preservation, either in liquid or frozen state.

Features of seminal plasma cytokine profile and an occurrence of pregnancy when carrying out a program of assisted reproductive technologies

Aim. To asses an effectiveness of assisted reproductive technologies (ART) program given characteristics of the cytokine profile of seminal plasma (SP) entering the female reproductive tract during sexual intercourse. Outcomes and methods. 33 married couples who applied for a treatment of infertility by means of in vitro fertilization /ICSI (Intracytoplasmic Sperm Injection) were included in the prospective study. Patients were recommended to have sexual intercourse with no restrictions during treatment and to have the last sexual intercourse 3 days before an intended transvaginal puncture (ITP). Testing of cytokines (transforming growth factor (TGF)-b1, interferon (IFN)-g, interleukin (IL)-33, IL-6, IL-8, IL-23, IL-10, tumor necrosis factor (TNF)-a, IL-18, IL- 17A, IFN-a, IL-12, monocyte chemotactic protein-1) levels in samples of partners’ SP obtained on the day having ITP were carried out using multiplex analysis with LegendPlex kits (BioLegend, USA). Results. When comparing a cytokine profile of SP in couples who did not become pregnant (n=25) and couples who become pregnant (n=8) increased IL-18 and reduced IL-10 levels (p=0,017 and p=0, 01 respectively) were revealed in the group which got pregnant. To assess a clinical relevance of cytokine content in SP ROC (Receiver Operating Characteristics) curve was used. It was established that determining of IL-18 concentration in SP has the greatest diagnostic significance (the area under a curve was 0.792±0.107, test sensitivity - 62.5%, test specificity - 95.24% at threshold concentration&gt;210.43 pg/ml). Incidence of pregnancy at IL-18 concentration above threshold levels was 83,3% while at lower concentrations of IL-18 pregnancy occurred only in 13.0% of women. Conclusions. Elevated IL-18 levels and decreased IL-10 levels in SP of female patients’ partners who don’t have restriction of sexual life when treating infertility with ART are favorable factors for a pregnancy to occur.

Study of products of IL-12 and IL-18 in sperm and blood of men, suffering on oligosymptomatic forms of a chronic urogenital infection

In order to increase the success of treating by family doctor male with infertility, developing the right tactics and optimal treatment regimens, verification of various factors of male infertility is required. From the accuracy of determining the causes of male infertility depends the further study of the pathogenesis of reproduction impairment, which will contribute to the development of effective therapies. More and more facts are being accumulated that point to the important role of immune processes in the pathogenesis of infertility, which provides the basis for studying the levels of cytokines in patients with chronic urogenital inflammatory processes. The objective : study the production of IL-12 and IL-18 cytokines in seminal plasma in males with low asymptomatic forms of chronic urogenital infection depending on fertility. Materials and methods . The patients (n=72) were examined for small asymptomatic forms of chronic inflammation of the urogenital tract, aged from 22 to 40 years. All surveyed are divided into two groups: the first (35 people) – infertile men and the second (37 people) – fertile men. Duration of infertility in patients of the І th group was 3 6 years. Polymerase chain reaction (PCR) method was used to detect Chlamidia trachomatis, M. hominis, U. urealyticum, V. herpes in seminal plasma. At the beginning of the survey, it took more than a year after the completion of the previous treatment. Sperm studies were performed and sperm morphology was determined using staining for Papanicolaou. Levels of IL-12 and IL-18 cytokines were studied in serum and in the semen. Control parameters of cytokine concentrations were obtained during the study of 24 fertile men without clinical signs of a disease of a similar age composition. Results . Investigation of cytokine levels in patients, depending on fertility, found that in the I group (male infertility), the level of IL-12 in the semen tended to decrease, with its concentration in blood in patients in this group did not differ from the norm. As for the group of fertile men (group II), an immunological study indicates diverse shifts in concentrations of IL-12 and IL-18 in both serum and seminal plasma. An increase in the content of the studied cytokines is much more pronounced in the semen than in the serum. The spermometry of both groups of patients showed that hyperfertility and microsomal type of sperma was observed in infertility men, and in fertile men, although there was a normospermia, however, in that part of the patients of the II group, which showed a decrease in the concentrations of IL-12 and IL-18 in the semen we stated an increase in the pathological forms of sperm. The data obtained indicate, in our opinion, the participation of IL-12 and IL-18 in the course of inflammation (mainly increased concentrations) and participation in the process of maturation of sperm. Conclusion . Microenvironment is important during maturation of the germ cells. An important role in this is played by cytokines, in particular IL-12 and IL-18. The study of the concentration of IL-12 and IL-18 in semen is more appropriate than in serum when the inflammatory process is established in the urogenital tract. Reduced concentrations of IL-12 and IL-18 in semen indicate the development of infertility, as evidenced by the presence of pathospermia. Immunocorrection therapy in men with oligosymptomatic forms of chronic inflammation of the urogenital tract complicated by infertility should take into account the shift of the cytokine profile both in serum and in the semen.

Clinical and Biochemical Influence of Prostatic Stones

Introduction: The study aimed to explore clinical influence of prostatic stones on lower urinary tract symptoms (LUTS), seminal plasma cytokines, and serum biomarkers. Materials and Methods: A total of 70 men aged ≤50 years with LUTS divided into 2 groups: group with stones (GSt) and group without prostatic stones (GNoSt). All subjects completed the International Prostate Symptom Score (IPSS) questionnaire and National Institutes of Health Chronic Prostatitis Symptom Index (NIH-CPSI) scoring questionnaire. Pre- and post-prostate massage test and uroflowmetry were performed. The serum concentration of total prostate specific antigen (PSA), free PSA, and free/total PSA (f/t PSA) ratio, seminal concentration of cytokines interleukin (IL)-1β, IL-6, IL-8, IL-10, IL-12p70, and tumor necrosis factor-alpha were measured. Results: GSt subjects had significantly more severe symptoms based on IPSS answers (p = 0.0289). All domains in NIH-CPSI scores were significantly higher in the GSt group: pain (p = 0.001), urinary symptoms (p = 0.023), quality of life (p = 0.008), and with overall (p = 0.003). GSt subjects also had significantly lower maximum urinary flow (Qmax; p = 0.011), lower f/t PSA ratio (p = 0.048), and higher concentration of IL-1β (p = 0.011) and IL-8 (p = 0.001). Conclusions: Prostatic stones may influence the severity of LUTS and the symptoms of chronic prostatitis. They might reduce Qmax rate and lead to reduction of the f/t PSA ratio and produce more severe inflammation causing increased seminal concentration of IL-1β and IL-8.

Seminal plasma transforming growth factor-β, activin A and follistatin fluctuate within men over time.

Do seminal plasma transforming growth factor-β (TGFB) cytokines vary within individuals over time, and does this relate to sperm parameters, age or prior abstinence? Activin A and follistatin, and to a lesser extent TGFB1, TGFB2 and TGFB3, vary within individuals over time, in association with duration of abstinence. Seminal plasma TGFB cytokines can influence sperm function and reproductive success through interactions with the female reproductive tract after coitus. Over time, individual sperm parameters fluctuate considerably. Whether seminal fluid TGFB cytokines vary similarly, and the determinants of any variance, is unknown. Between two and seven semen samples were collected from each of 14 fertile donors at 6-10 week intervals over the course of 12 months, then seminal plasma cytokines and sperm parameters were measured. The concentrations and total amounts per ejaculate of TGFB1, TGFB2, TGFB3, activin A and follistatin were determined using commercial assays. Sperm parameters were assessed according to WHO IV standards. Mixed model analysis was utilised to determine the relative contribution of between- and within-individual factors to the variance. Relationships between cytokines and sperm parameters, as well as effect of age and duration of abstinence, were investigated by correlation analysis. Within-individual variability contributed to the total variance for all cytokines and sperm parameters, and was a stronger determinant than between-individual variability for activin A and follistatin as well as for total sperm concentration and sperm motility. Positive correlations between each of the three TGFB isoforms, and activin and follistatin, suggest co-regulation of synthesis. Duration of abstinence influenced total content of TGFB1, TGFB2, activin A and follistatin. TGFB1 correlated inversely with age. A limited number of donors from a single clinic were investigated. Clinical information on BMI, nutrition, smoking and other lifestyle factors was unavailable. Further studies are required to determine whether the findings can be generalised to larger populations and different ethnicities. These data reveal substantial variation over time in seminal fluid cytokines and indicate that repeated analyses are required to gain precise representative data on an individual's status. Within-individual variation in seminal fluid components should be taken into account when investigating seminal fluid cytokines. This study was supported by grants from the National Health and Medical Research Council of Australia, ID453556 and APP1041332. The authors have no competing interests to disclose.

Environmental exposure to lead induces oxidative stress and modulates the function of the antioxidant defense system and the immune system in the semen of males with normal semen profile

We investigated the associations between environmental exposure to lead and a repertoire of cytokines in seminal plasma of males with normal semen profile according to the WHO criteria. Based on the median lead concentration in seminal plasma, 65 samples were divided into two groups: low (LE) and high exposure to lead (HE). Differences in semen volume and the pH, count, motility and morphology of sperm cells were not observed between the examined groups. The total oxidant status value and the level of protein sulfhydryl groups as well as the activities of manganese superoxide dismutase and catalase were significantly higher in the HE group, whereas the total antioxidant capacity value and the activities of glutathione reductase and glutathione-S-transferase were depressed. IL-7, IL-10, IL-12, and TNF-α levels were significantly higher in the HE group compared with the LE group. Environmental exposure to lead is sufficient to induce oxidative stress in seminal plasma and to modulate antioxidant defense system.

Seminal Plasma HIV-1 RNA Concentration Is Strongly Associated with Altered Levels of Seminal Plasma Interferon-γ, Interleukin-17, and Interleukin-5

Seminal plasma HIV-1 RNA level is an important determinant of the risk of HIV-1 sexual transmission. We investigated potential associations between seminal plasma cytokine levels and viral concentration in the seminal plasma of HIV-1-infected men. This was a prospective, observational study of paired blood and semen samples from 18 HIV-1 chronically infected men off antiretroviral therapy. HIV-1 RNA levels and cytokine levels in seminal plasma and blood plasma were measured and analyzed using simple linear regressions to screen for associations between cytokines and seminal plasma HIV-1 levels. Forward stepwise regression was performed to construct the final multivariate model. The median HIV-1 RNA concentrations were 4.42 log10 copies/ml (IQR 2.98, 4.70) and 2.96 log10 copies/ml (IQR 2, 4.18) in blood and seminal plasma, respectively. In stepwise multivariate linear regression analysis, blood HIV-1 RNA level (p<0.0001) was most strongly associated with seminal plasma HIV-1 RNA level. After controlling for blood HIV-1 RNA level, seminal plasma HIV-1 RNA level was positively associated with interferon (IFN)-γ (p=0.03) and interleukin (IL)-17 (p=0.03) and negatively associated with IL-5 (p=0.0007) in seminal plasma. In addition to blood HIV-1 RNA level, cytokine profiles in the male genital tract are associated with HIV-1 RNA levels in semen. The Th1 and Th17 cytokines IFN-γ and IL-17 are associated with increased seminal plasma HIV-1 RNA, while the Th2 cytokine IL-5 is associated with decreased seminal plasma HIV-1 RNA. These results support the importance of genital tract immunomodulation in HIV-1 transmission.

The Effects of Honey Supplementation on Seminal Plasma Cytokines, Oxidative Stress Biomarkers, and Antioxidants During 8 Weeks of Intensive Cycling Training

The purpose of this study was to examine the effects of natural honey supplementation on seminal plasma cytokines, oxidative stress biomarkers, and antioxidants during 8 weeks of intensive cycling training in male road cyclists. Thirty-nine healthy nonprofessional male road cyclists aged 18-28 years participated in this study. The participants were randomly assigned to exercise + supplement (E + S, n = 20) and exercise (E, n = 19) groups. All subjects participated in 8 weeks of intensive cycling training. Ninety minutes before each training session, subjects in the E + S group supplemented with 70 g of honey, whereas subjects in the E group received 70 g of an artificial sweetener. All subjects had an initial semen sampling at baseline (T(1)). The next 6 semen collections were collected immediately (T(2)) and 12 (T(3)) and 24 hours (T(4)) after the last training session in week 4, as well as immediately (T(5)) and 12 (T(6)) and 24 hours (T(7)) after the last training session in week 8, respectively. In the E group, 8 weeks of intensive cycling training significantly increased seminal interleukin (IL)-1β, IL-6, IL-8, tumor necrosis factor (TNF)-α, reactive oxygen species (ROS), and malondialdehyde (MDA) levels (P < .008) and significantly decreased the levels of seminal superoxide dismutase (SOD), catalase, and total antioxidant capacity (TAC) (P < .008). Significantly less elevation in seminal IL-1β, IL-6, IL-8, TNF-α, ROS, and MDA levels (P < .008) and significant increases in seminal SOD, catalase, and TAC concentrations were observed after the honey supplementation in the E + S group (P < .008). It may be possible that honey supplementation following long-term intensive cycling training would be effective in attenuating the probable aggravating effects of intensive cycling training on spermatogenesis and fertility capacity in road cyclists.

Cytokine expression in the seminal plasma and its effects on fertilisation rates in an IVF cycle

Cytokines are released by various immunocompetent cell subsets in the male urogenital tract and are thought to affect sperm cell function and reproductive process. The aim of the study was to evaluate the levels and a possible role of seven seminal plasma cytokines with fertilisation rates in men attending an in vitro fertilisation (IVF) programme. A total of 36 men of couples who were undergoing traditional IVF in a regional reproductive medicine unit were recruited into this prospective study. Cytokines such as interleukin (IL)-6, IL-8, IL-10, IL-11, IL-12, tumour necrosis factor alpha (TNF-α) and interferon gamma (IFN-γ) in the seminal plasma were determined using enzyme linked immunosorbent assay. IL-6, IL-8, IL-10, IL-11 and IFN-γ were detected in all samples. IL-12, and TNF-α were detected in most samples. Levels of IL-11 were significantly higher in the good fertiliser group (P ≤ 0.05). Positive correlation between cytokines such as IL-6 and IL-8 (P < 0.03), IL-10 and IL-11 (P < 0.001) and IFN-γ and IL-10 and IL-11 (P < 0.04 and P < 0.0001 respectively) were found. Our study confirms that the six cytokines other than IL-11 do not affect spermatozoon-oocyte interaction and fertilisation rates in IVF. IL-11 could have a role in the fertilising capacity of the spermatozoa. Significant correlation exists among these cytokines which shows that cytokines rarely act in isolation but rather in a network.