Weak association of anti-sperm antibodies and strong association of familial cryptorchidism/infertility with HLA-DRB1polymorphisms in prepubertal Ukrainian boys

<p>Abstract</p> <p>Background</p> <p>Cryptorchidism is a frequent syndrome occurring in 1-2% of males within the first year of age. Autoimmune reactions, particularly directed to testicular elements and/or spermatozoa have been found to be often associated with cryptorchidism. Therefore we investigated in this study the frequency of HLA class II alleles in order to recognize possible genetic predisposition for antisperm antibodies development in prepubertal boys with diagnosed cryptorchidism in Caucasoid population.</p> <p>Methods</p> <p>Sixty prepubertal boys with cryptorchidism and sixty healthy boys were examined for anti-sperm antibodies by indirect immunobead test as well as for their <it>HLA-DRB1 </it>and -<it>DQB1 </it>alleles using DNA obtained from peripheral blood leukocytes. The typing of <it>HLA-DRB1 </it>and -<it>DQB1 </it>was performed by using PCR-SSP low resolution method.</p> <p>Results</p> <p>Allele frequencies of <it>HLA-DRB1 </it>and <it>HLA-DQB1 </it>did not differ between boys with cryptorchidism and control boys. However, weakly significant differences in <it>DRB1*04 </it>(<it>p corrected </it>= 0.0475) and <it>DQB1*06 </it>(<it>p corrected </it>= 0.0385) were seen between cryptorchid patients with and without AsA, but none of these two patient groups differed significantly in <it>HLA </it>class II frequencies from controls except for AsA-negatives and <it>HLA-DQB1*06 </it>(<it>p corrected </it>= 0.0247). On the other hand, comparison of cryptorchid boys with familial cryptorchidism and/or infertility to control boys revealed highly significant (<it>p corrected </it>= 0.0006) difference in <it>HLA-DRB*11 </it>frequency, whereas boys with sporadic cryptorchidism did not differ from control. A much weaker, but still significant difference in <it>DRB*11 </it>frequency was also observed between boys with bilateral cryptorchidism and controls (<it>p corrected </it>= 0.037), whereas patients with unilateral cryptorchidism were not different from control in frequency of any <it>HLA-DRB1 </it>or -<it>DQB1 </it>allele tested.</p> <p>Conclusions</p> <p>Predisposition to produce anti-sperm antibodies seems to be only weakly associated with <it>HLA </it>class II genes, although this question requires further study on much larger population sample. It is plausible that familial and sporadic cryptorchidism may present distinct genetic background. The same may, to lower extent, apply to bilateral and unilateral cryptorchidism.</p

Flow cytometry application in the evaluation of antisperm antibodies in sera samples of infertile people and prepubertal boys with gonadal disorders

Abstract Objectives: The aim of the following study was to assess antisperm antibodies in sera samples of infertile men and women, as well as from prepubertal boys by means of flow cytometry. Material and methods: We tested sera samples of infertile and fertile adult populations, prepubertal boys with gonadal disorders and healthy prepubertal boys. The indirect immunobead test and flow cytometry were used to detect antisperm antibodies. Results: The comparison of antisperm antibody levels in sera samples of adult infertile versus healthy controls (men and women) evaluated by means of flow cytometry did not reveal statistically significant differences. The only significant correlation found were results obtained by IDIBT and FCM for IgG antisperm antibodies for infertile adult group (r=0.507, p=0.012). The comparison of antisperm antibody levels in sera samples from prepubertal boys revealed statistically significant differences for all tested antibody isotypes. Diagnostic values compared for both assays showed markedly better discriminatory ability of flow cytometry for analyzed groups of prepubertal boys than for adult populations. Conclusions: Flow cytometry test may be used to verify antisperm antibody levels in prepubertal boys with testicular failures

Global 5mC and 5hmC DNA Levels in Human Sperm Subpopulations with Differentially Protaminated Chromatin in Normo- and Oligoasthenozoospermic Males

Epigenetic modifications play a special role in the male infertility aetiology. Published data indicate the link between sperm quality and sperm chromatin protamination. This study aimed to determine the relationship between methylation (5mC) and hydroxymethylation (5hmC) in sperm DNA, with respect to sperm chromatin protamination in three subpopulations of fertile normozoospermic controls and infertile patients with oligo-/oligoasthenozoospermia. For the first time, a sequential staining protocol was applied, which allowed researchers to analyse 5mC/5hmC levels by immunofluorescence staining, with a previously determined chromatin protamination status (aniline blue staining), using the same spermatozoa. TUNEL assay determined the sperm DNA fragmentation level. The 5mC/5hmC levels were diversified with respect to chromatin protamination status in both studied groups of males, with the highest values observed in protaminated spermatozoa. The linkage between chromatin protamination and 5mC/5hmC levels in control males disappeared in patients with deteriorated semen parameters. A relationship between 5mC/5hmC and sperm motility/morphology was identified in the patient group. Measuring the 5mC/5hmC status of sperm DNA according to sperm chromatin integrity provides evidence of correct spermatogenesis, and its disruption may represent a prognostic marker for reproductive failure

Seminal Plasma Analysis of Oxidative Stress in Different Genitourinary Topographical Regions Involved in Reproductive Tract Disorders Associated with Genital Heat Stress

The pathophysiological mechanisms responsible for male subfertility/infertility caused by or complicated by genital heat stress remains unclear in many respects. Because seminal plasma creates the environment for the proper functioning of spermatozoa, in this study, we verified the associations among standard spermiograms, seminal biochemical parameters (neutral alpha-glucosidase, fructose, and citric acid) and oxidative stress markers (total antioxidant capacity, catalase activity, superoxide dismutase activity, and malondialdehyde concentration) in distinct entities associated with male infertility with and without long-time exposure to local hyperthermia. We demonstrated that men exposed to prolonged environmental or clinically recognized local heat stress in adulthood may suffer from dysregulation of seminal antioxidant components, which can be directly associated with epididymal and prostate function. The comparative analysis of the studied parameters showed numerous correlations among all biochemical parameters (particularly neutral alpha-glucosidase) with low standard semen quality in almost all the investigated infertile groups. In light of the data obtained in this originally designed study, we conclude that more attention should be paid to the epididymis and accessory gland function in subfertile and infertile men exposed to genital heat stress, especially in the context of novel treatment algorithms (targeted therapies)