Screening of sperm antigen epitopes by phage display technique and its preliminary clinical application.

BACKGROUND: At present, there is a lack of standardized preparation methods of sperm antigen for the detection of antisperm antibody (AsAb). To screen sperm antigen mimotopes from a phage display random peptide library and use them to establish an enzyme-linked immunosorbent assay (ELISA) for the detection of AsAb, immunoglobulins were extracted from the sera of rabbits with positive AsAb and negative AsAb, respectively, by the saturated ammonium sulfate method, and a phage display 12-mer peptide library was affinity panned by the extracted immunoglobins coated on the ELISA plate. Then, the obtained positive phage clones were identified by ELISA and sent for sequencing and peptides synthesis. Last, a diagnostic ELISA was established to detect clinical serum and seminal plasma samples. RESULTS: A total of sixty phage clones were chosen by affinity panning, and sixteen of them reacted positively with AsAb in indirect ELISA and sandwich ELISA. Following DNA sequencing and translation, the peptide sequences of the sixteen positive clones were obtained. By comparison in Blast database, four of sixteen positive clones were found to be closely related to male reproduction. Two (#1 and #25) of four mimotopes were synthesized, and an ELISA method was established using the two mimotopes as sperm specific antigens. One hundred and thirty-four serum samples and seventy-four seminal plasma samples from infertile couples were analyzed by the established ELISA with #1 and #25 mimotopes, respectively. The positive rates of AsAb in serum samples were 20.15% (27/134) for #1 and 11.19% (15/134) for #25, respectively, and the coincidence rate between them was 91.04% (122/134). The positive rates of AsAb in seminal plasma samples were 1.35% (1/74) for both #1 and #25, and the coincidence rate was 100%. CONCLUSION: Sperm antigen mimotopes can be obtained successfully by the phage display technique, and can be used as standard sperm specific antigens to establish an ELISA method for the detection of AsAb.

RéSUMé: CONTEXTE: À ce jour, il n'existe pas de méthodes normalisées de préparation d'antigènes spermatiques pour la détection des anticorps anti-spermatozoïdes (ACAS). Dans le but d'élaborer un tel test ELISA (enzyme-linked immunosorbent assay), nous avons extrait de sérum de lapins des anticorps anti-spermatozoïdes humains via la technique du sulfate d'ammonium saturé et en ayant recours à une librairie phagique de peptides (12-mer). Les clones positifs ont été identifiés par ELISA, séquencés à façon et les peptides correspondants ont été synthétisés. In fine, un test ELISA diagnostic a été conçu pour être utilisé avec des échantillons cliniques de sérum et de plasmas séminaux. RéSULTATS: Au total, soixante clones de phages ont été sélectionnés, et seize d'entre eux se sont avérés interagir avec les ACAS en ELISA indirect comme en ELISA sandwich. Les séquences peptidiques de ces seize clones positifs ont été obtenues. Par comparaison avec les bases de données (Blast), quatre de ces seize clones positifs se sont révélés être étroitement liés à la reproduction masculine. Deux des quatre mimotopes (#1 et #25) ont été synthétisés, et un test ELISA a été généré en utilisant ces deux mimotopes comme antigènes spécifiques des spermatozoïdes. Cent trente-quatre échantillons de sérum et soixante-quatorze échantillons de plasma séminal de patients de couples infertiles ont alors été analysés avec ce test ELISA. Respectivement, les échantillons sériques se sont révélés positifs à 20,15% (27/134) pour le mimotope #1 et à 11,19% (15/134) pour le mimotope #25, avec un taux de coïncidence de 91,04% (122/134). Seul un échantillon de plasma séminal (1/74, soit 1, 35%) s'est révélé positif à la fois pour le mimotope #1 et #25 (coïncidence 100%). CONCLUSION: La technique « phage display¼ nous a permis d'identifier des mimotopes d'antigènes spermatiques qui ont pu être utilisés afin de générer un test ELISA pour la détection d'anticorps anti-spermatozoïdes.

Multifunctional Metal-Organic Framework Exoskeletons Protect Biohybrid Sperm Microrobots for Active Drug Delivery from the Surrounding Threats.

Utilizing spermatozoa as the engine unit of robotic systems at a microscale has brought revolutionized inspirations and strategies to the biomedical community. However, the motility of sperms is impaired by the surrounding threats. For example, the antisperm antibody (AsA) can specifically bind with surface antigens on the sperm membrane and adversely affect their propulsion, hindering the operation of sperm-based microrobots in practical environments. In the present work, we report a biohybrid sperm microrobot by encapsulating sperm cells within metal-organic frameworks (MOFs) and zeolitic imidazolate framework-8 (ZIF-8) nanoparticles (NPs) (ZIFSpermbot), capable of active drug delivery and cytoprotection from the biological threats of AsA. ZIF-8 NPs can be facilely coated on the sperm membrane through complexation with tannic acid. Such cell surface engineering has a negligible impact on sperm motility under optimized conditions. The selective permeability of the resulting porous ZIF-8 wrappings protects ZIFSpermbots from the specific binding of AsA, enabling the preservation of intrinsic propulsion of the sperm engine. Besides, ZIF-8 wrappings sustainably release zinc ions and attenuate the oxidative damage generated in sperm cells, allowing the maintenance of sperm movement. Combining the effective protection of sperm propulsion with the drug-loading capacity of ZIF-8 NPs provides new applicability to ZIFSpermbots in risky surroundings with AsA, exhibiting rapid migration in a microfluidic device for active drug delivery with enhanced therapeutic efficacy due to their retained effective propulsion. Imparting bioengine-based microrobots with multifunctional wrappings holds great promise for designing adaptive cell robots that endure harsh environments toward locally extended and diverse operations, facilitating their use in practical and clinical applications.

Sperm immobilization test and quantitative sperm immobilization test using frozen-thawed sperm preparation applied with computer-aided sperm analysis.

PURPOSE: In a previous study, a new method was described using the sperm immobilization test (SIT) with computer-aided sperm analysis (CASA). However, obtaining high-quality sperm as needed was a known issue. Here, we compared the results of using frozen-thawed sperm and fresh sperm for the SIT using the CASA method. METHODS: For the frozen-thawed preparation, 500 µL of condensed semen and 500 µL of Sperm Freeze were mixed in a cryovial and cryopreserved in liquid nitrogen. Density gradient centrifugation was used for the collection of motile sperm in both the fresh and frozen-thawed sperm preparations. A total of 50 serum samples were prepared for both the fresh and frozen-thawed sperm with each sample tested containing 10 µL of serum, 1 µL of either fresh or frozen motile sperm suspension, and 2 µL of complement. Sperm motilities were measured using CASA after a 1-hour incubation period for both fresh and frozen-thawed sperm. RESULTS: Both fresh and frozen-thawed sperm reacted similarly when exposed to serum containing sperm-immobilizing antibodies asserting the use of frozen-thawed sperm for the diagnosis of immunological infertility. CONCLUSION: These results suggest the possibility of using cryopreserved sperm for the SIT when fresh sperm is unavailable.

Production and characterization of a human antisperm monoclonal antibody against CD52g for topical contraception in women.

BACKGROUND: Approximately 40% of human pregnancies are unintended, indicating a need for more acceptable effective contraception methods. New antibody production systems make it possible to manufacture reagent-grade human monoclonal antibodies (mAbs) for clinical use. We used the Nicotiana platform to produce a human antisperm mAb and tested its efficacy for on-demand topical contraception. METHODS: Heavy and light chain variable region DNA sequences of a human IgM antisperm antibody derived from an infertile woman were inserted with human IgG1 constant region sequences into an agrobacterium and transfected into Nicotiana benthamiana. The product, an IgG1 mAb ["Human Contraception Antibody" (HCA)], was purified on Protein A columns, and QC was performed using the LabChip GXII Touch protein characterization system and SEC-HPLC. HCA was tested for antigen specificity by immunofluorescence and western blot assays, antisperm activity by sperm agglutination and complement dependent sperm immobilization assays, and safety in a human vaginal tissue (EpiVaginal™) model. FINDINGS: HCA was obtained at concentrations ranging from 0.4 to 4 mg/ml and consisted of > 90% IgG monomers. The mAb specifically reacted with a glycan epitope on CD52g, a glycoprotein produced in the male reproductive tract and found in abundance on sperm. HCA potently agglutinated sperm under a variety of relevant physiological conditions at concentrations ≥ 6.25 µg/ml, and mediated complement-dependent sperm immobilization at concentrations ≥ 1 µg/ml. HCA and its immune complexes did not induce inflammation in EpiVaginal™ tissue. INTERPRETATION: HCA, an IgG1 mAb with potent sperm agglutination and immobilization activity and a good safety profile, is a promising candidate for female contraception. FUNDING: This research was supported by grants R01 HD095630 and P50HD096957 from the National Institutes of Health.

Distribution of Human Papillomavirus and Antisperm Antibody in Semen and Its Association with Semen Parameters Among Infertile Men.

BACKGROUND: Sexually transmitted infections (STIs) can be associated with infertility. Human papillomavirus (HPV) has been identified as a potential agent in male infertility. Also, anti-sperm antibodies (ASA) have been detected in men with infertility. The aim of this study was to investigate the prevalence and association of HPV and ASA in infected semen of infertile men. METHODS: This cross-sectional study was performed on 96 infertile men referring to infertility treatment center of Kashan University of Medical Sciences during March 2017 till September 2017 in Iran. Semen analysis and diagnostic PCR test were performed for detection of HPV DNA. The semen parameters in HPV infected and ASA positive samples were compared with HPV non-infected and ASA negative samples. Chi square test was used to determine the correlation between variables and p<0.05 was considered statistically significant. RESULTS: HPV DNA and ASA were detected in 17.4% and 15.2% of 96 semen samples, respectively. Semen volume, sperm count, sperm motility and the normal morphology rate were significantly decreased in HPV-positive subjects (p=0.004, p= 0.016, p<0.001, and p=0.017, respectively). Also, sperm motility was significantly decreased in ASA-positive subjects (p=0.002), also patients with HPV infection had a higher rate of ASA than the non-HPV group. In contrast to ASA, HPV infection had a significant correlation with education level (p=0.039). CONCLUSION: The findings suggest that asymptomatic seminal infection of HPV and ASA by adversely affecting sperm quality, in particular sperm motility and count, may play an important role in male infertility.

Immune regulatory molecules as modifiers of semen and fertility: A review.

Declining fertility rates in both human and animals is a cause for concern. While many of the infertility cases are due to known causes, idiopathic infertility is reported in 30% of the infertile couples. In such cases, 18% of the infertile males carry antisperm antibodies (ASAs). Such data are lacking in livestock, wherein 20-30% of the animals are being culled due to low fertility. In males, the blood-testis barrier (BTB) and biomolecules in the semen provide an immuno-tolerant microenvironment for spermatozoa as they traverse the immunologic milieu of both the male and female reproductive tracts. For example, insults from environmental contaminants, infections and inflammatory conditions are likely to impact the immune privilege state of the testis and fertility. The female mucosal immune system can recognize allogenic spermatozoa-specific proteins affecting sperm kinematics and sperm-zona binding leading to immune infertility. Elucidating the functions and pathways of the immune regulatory molecules associated with fertilization are prerequisites for understanding their impact on fertility. An insight into biomolecules associated with spermatozoal immune tolerance may generate inputs to develop diagnostic tools and modulate fertility. High-throughput sequencing technologies coupled with bioinformatics analyses provides a path forward to define the array of molecules influencing pregnancy outcome. This review discusses the seminal immune regulatory molecules from their origin in the testis until they traverse the uterine environment enabling fertilization and embryonic development. Well-designed experiments and the identification of biomarkers may provide a pathway to understand the finer details of reproductive immunology that will afford personalized therapies.

[Association of Ureaplasma urealyticum with the types of antisperm antibody in infertile men].

OBJECTIVE: To investigate the prevalence of Ureaplasma urealyticum (UU) infection in infertile men, its influence on routine semen parameters and the distribution of antisperm antibody (AsAb) and its types in infertile patients with UU infection. METHODS: We detected the positive rate of UU infection, semen parameters, and the distribution of AsAb and its types in 662 infertile men and 25 normal fertile male controls followed by comparison of the obtained data between the two groups of subjects. RESULTS: The positive rate of UU infection was significantly higher in the infertile men than in the normal controls (52.87% ï¼»350/662ï¼½ vs 16.00% ï¼»4/25ï¼½, χ2 = 11.68, P <0.05). The semen volume, sperm count, sperm concentration and percentage of progressively motile sperm were remarkably lower in the UU-positive infertile males than in the control group (P <0.05). No statistically significant difference was observed between the UU-positive and UU-negative groups in the positive rates of total AsAb (43.4% vs 36.5%, χ2 = 3.25, P >0.05) and AsAb IgA, IgM and IgG in the seminal plasma, or in the percentages of serum AsAb IgM (16.9% vs 20.5%, χ2 = 1.22, P >0.05) and IgG (32.7% vs 28.9%, χ2 = 0.99, P >0.05) except in that of serum AsAb IgA (23.6% vs 17.0%, χ2 = 4.03, P <0.05). CONCLUSIONS: The UU infection rate is high in infertile males, which decreases the semen volume, total sperm count, motile sperm concentration and percentage of progressively motile sperm and increases the positive rate of serum AsAb IgA.

Association of Ureaplasma urealyticum with the types of antisperm antibody in infertile men / 中华男科学杂志

Objective@#To investigate the prevalence of Ureaplasma urealyticum (UU) infection in infertile men, its influence on routine semen parameters and the distribution of antisperm antibody (AsAb) and its types in infertile patients with UU infection.@*METHODS@#We detected the positive rate of UU infection, semen parameters, and the distribution of AsAb and its types in 662 infertile men and 25 normal fertile male controls followed by comparison of the obtained data between the two groups of subjects.@*RESULTS@#The positive rate of UU infection was significantly higher in the infertile men than in the normal controls (52.87% ［350/662］ vs 16.00% ［4/25］, χ2 = 11.68, P 0.05) and AsAb IgA, IgM and IgG in the seminal plasma, or in the percentages of serum AsAb IgM (16.9% vs 20.5%, χ2 = 1.22, P >0.05) and IgG (32.7% vs 28.9%, χ2 = 0.99, P >0.05) except in that of serum AsAb IgA (23.6% vs 17.0%, χ2 = 4.03, P <0.05).@*CONCLUSIONS@#The UU infection rate is high in infertile males, which decreases the semen volume, total sperm count, motile sperm concentration and percentage of progressively motile sperm and increases the positive rate of serum AsAb IgA.

[Impacts of different procedures of testicular sperm retrieval on testicular function and antisperm antibodies in azoospermia patients].

OBJECTIVE: To investigate the influence of different procedures of testicular sperm retrieval on the levels of serum inhibin B (INHB), antisperm antibodies (AsAb), follicle-stimulating hormone (FSH), and testosterone (T) in patients with azoospermia. METHODS: We randomly assigned 210 azoospermia patients to receive testicular sperm extraction (TESE, n = 50), testicular sperm aspiration (TESA, n = 56), testicular fine needle aspiration (TEFNA, n = 64), or microscopic TESE (micro-TESE, n = 40). We measured the levels of serum INHB, FSH, and T and the positive rate of AsAb before and at 1 and 3 months after surgery. RESULTS: Compared with the baseline, the levels of serum FSH at 1 and 3 months after surgery showed no statistically significant differences in the TESE (ï¼»8.51 ± 4.34ï¼½ vs ï¼»8.76 ± 3.07ï¼½ and ï¼»7.24 ± 3.32ï¼½ IU/L, P >0.05), TESA (ï¼»7.70 ± 2.72ï¼½ vs ï¼»7.90 ± 4.57ï¼½ and ï¼»8.04 ± 3.65ï¼½ IU/L, P >0.05), TEFNA (ï¼»6.04 ± 3.17ï¼½ vs ï¼»6.08 ± 2.70ï¼½ and ï¼»6.10 ± 3.32ï¼½ IU/L, P >0.05), or micro-TESE group (ï¼»6.59 ± 2.74ï¼½ vs ï¼»6.89 ± 1.78ï¼½ and ï¼»6.75 ± 2.57ï¼½ IU/L, P >0.05); the positive rate of AsAb (IgM) was significantly increased at 1 month in the TESE (0.00 vs 14.00%, P <0.05) and micro-TESE groups (2.50% vs 15.00%, P <0.05), while the serum T level markedly decreased in the two groups (ï¼»16.52 ± 6.25ï¼½ vs ï¼»9.25 ± 5.76ï¼½ nmol/L and ï¼»14.16 ± 5.45ï¼½ vs ï¼»8.23 ± 4.12ï¼½ nmol/L, P <0.05); the levels of serum INHB were remarkably reduced at 1 and 3 months in the TESE (ï¼»70.56 ± 23.17ï¼½ vs ï¼»42.63 ± 15.34ï¼½ and ï¼»44.05 ± 18.47ï¼½ pg/ml, P <0.05), TESA (ï¼»68.71 ± 14.74ï¼½ vs ï¼»40.55 ± 20.51ï¼½ and ï¼»42.11 ± 19.34ï¼½ pg/ml, P <0.05), TEFNA (ï¼»76.81 ± 27.04ï¼½ vs ï¼»46.31 ± 19.28ï¼½ and ï¼»48.32 ± 20.54ï¼½ pg/ml, P <0.05), and micro-TESE groups (ï¼»74.74 ± 28.35ï¼½ vs ï¼»45.27 ± 18.83ï¼½ and ï¼»47.64 ± 28.34ï¼½ pg/ml, P <0.05), but with no statistically significant differences among the four groups (P >0.05). CONCLUSIONS: Different procedures of testicular sperm retrieval have different impacts on the testicular function and AsAb in patients with azoospermia.

Expression and function of HSP110 family in mouse testis after vasectomy / 亚洲男科学杂志(英文版)

HSP110 functions to protect cells, tissues, and organs from noxious conditions. Vasectomy induces apoptosis in the testis; however, little is known about the reason leading to this outcome. The aim of the present study was to evaluate the expression and function of HSP110 in mouse testis after vasectomy. Following bilateral vasectomy, we used fluorescent Terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) to detect apoptosis, Western blotting and immunohistochemistry to examine HSP110 expression and localization. Serum antisperm antibody (AsAb) and testosterone were measured by Enzyme-linked immunosorbent assay (ELISA) and radioimmunoassay, respectively. Expression of endoplasmic reticulum stress (ERS) sensors and downstream signaling components was measured by Reverse Transcription-Polymerase Chain Reaction (RT-PCR), and the phosphorylation of eIF2 and JNK was detected by Western blotting. Vasectomy induced morphologic changes, increased apoptosis in the testis, increased serum AsAb, and decreased testosterone levels. After vasectomy, ORP150 mRNA level was increased first and then decreased, Bcl-2 was decreased, and the expression of HSPA4l, GRP78, GADD153, PERK, ATF6, IRE-1, XBP-1s, Bax, Bak, and caspases and the phosphorylation of eIF2 and JNK were increased. We present that an ER stress-mediated pathway is activated and involved in apoptosis in the testis after vasectomy. HSPA4l and ORP150 may play important roles in maintaining the normal structure and function of testis.

Impacts of different procedures of testicular sperm retrieval on testicular function and antisperm antibodies in azoospermia patients / 中华男科学杂志

Objective@#To investigate the influence of different procedures of testicular sperm retrieval on the levels of serum inhibin B (INHB), antisperm antibodies (AsAb), follicle-stimulating hormone (FSH), and testosterone (T) in patients with azoospermia.@*METHODS@#We randomly assigned 210 azoospermia patients to receive testicular sperm extraction (TESE, n = 50), testicular sperm aspiration (TESA, n = 56), testicular fine needle aspiration (TEFNA, n = 64), or microscopic TESE (micro-TESE, n = 40). We measured the levels of serum INHB, FSH, and T and the positive rate of AsAb before and at 1 and 3 months after surgery.@*RESULTS@#Compared with the baseline, the levels of serum FSH at 1 and 3 months after surgery showed no statistically significant differences in the TESE (［8.51 ± 4.34］ vs ［8.76 ± 3.07］ and ［7.24 ± 3.32］ IU/L, P >0.05), TESA (［7.70 ± 2.72］ vs ［7.90 ± 4.57］ and ［8.04 ± 3.65］ IU/L, P >0.05), TEFNA (［6.04 ± 3.17］ vs ［6.08 ± 2.70］ and ［6.10 ± 3.32］ IU/L, P >0.05), or micro-TESE group (［6.59 ± 2.74］ vs ［6.89 ± 1.78］ and ［6.75 ± 2.57］ IU/L, P >0.05); the positive rate of AsAb (IgM) was significantly increased at 1 month in the TESE (0.00 vs 14.00%, P 0.05).@*CONCLUSIONS@#Different procedures of testicular sperm retrieval have different impacts on the testicular function and AsAb in patients with azoospermia.

Value of detections of antinuclear antibody combined with anti-cardiolipin antibody, anti-sperm antibody and anti-β2-glycoprotein I antibody in female infertility / 国际检验医学杂志

Objective To investigate the value of detections of anti‐nuclear antibody (ANA) combined with anti‐cardiolipin anti‐body (ACA) ,anti‐sperm antibody (AsAb) and anti‐beta 2 glycoprotein I (β2 GPI) antibody in the diagnosis of female infertility dis‐ease .Methods A total of 187 female cases of infertility (infertility group) were detected serum ANA and AsAb by the indirect im‐munofluorescence assay ,and ACA andβ2 GPI antibody by the enzyme linked immunosorbent assay (ELISA) .ANA ,ACA ,AsAb andβ2 GPI antibody also were detected in 80 females cases of normal fertility (normal group) .Results Among 187 cases of female infer‐tility ,ANA positive rate was 18 .1% (34/187) and which in the normal group was 2 .5% (2/80) .The ACA positive rate was 22 .3%(43/187) in the infertility group and 5 .0% (4/80) in the normal group ;the AsAb positive rate was 18 .7% (35/187) in the infertil‐ity group and 3 .8% (3/80) in the normal group ;theβ2 GPI positive rate was 20 .3% (38/187) in the infertility group and 3 .8% (3/80) in the normal group;the differences between the two groups had statistical significance (P<0 .05) .Conclusion Infertility is closely correlated with the in vivo existence of ANA ,ACA ,AsAb andβ2 GPI antibody ,the joint detection is conducive to find the e‐tiology of infertility and improve the clinical diagnosis rate .

Antisperm antibodies in infertile men and their effect on semen parameters: a systematic review and meta-analysis.

BACKGROUND: Antisperm antibodies (ASA) in males cause the autoimmune disease 'immune infertility'. The mechanism of ASA cause male infertility is not clear. Present studies have investigated the effect of ASA and their incidence in men with unexplained infertility, as well as to evaluate the correlation between the ASA and semen parameter alterations but have shown inconsistent results. We performed a systematic literature review and meta-analysis to assess the association between ASA and basic semen parameters in infertility men. METHODS: Systematic literature searches were conducted with PubMed, EMBASE, Science Direct/Elsevier, CNKI and the Cochrane Library up to October 2014 for case-control studies that involved the impact of ASA on semen parameters. Meta-analysis was performed with Review Manager. Standard mean differences (SMD) of semen parameters were identified with 95% CI in a random or fixed effects model. RESULTS: Eight studies were identified, including 238 cases of ASA positive infertility male and 929 ASA negative controls. Our results illustrated that the sperm concentration and sperm motility (a+b) from ASA positive patients were significantly lower than ASA negative controls (SMD (95% CI) -23.64 [-43.47, -3.81], -16.40 [-27.92, -4.88], respectively). However, semen liquefaction time in the ASA positive group was significantly longer than the control group (SMD (95% CI) 4.19 [1.72, 6.66]). There was no significant effect of ASA on the sperm volume, sperm viability, sperm progressive motility, sperm normal morphology and sperm abnormal morphology. CONCLUSIONS: The present study illustrates that there was a significant negative effect of ASA on sperm concentration, sperm motility (a+b) and sperm liquefaction.

Study of the Changes of Acrosomal Enzyme, Nitric Oxide Synthase, and Superoxide Dismutase of Infertile Patients with Positive Antisperm Antibody in Seminal Plasma.

The purpose of this article is to study the effect of positive antisperm antibody (AsAb) in seminal plasma on acrosomal enzyme activity, nitric oxide synthase (NOS) activity, and superxide dismutase (SOD) activity of spermatozoa. A total of 40 infertility patients with positive AsAb in seminal plasma were selected as experimental group, and 40 fertile males were selected as control group. Based on the changes in absorbance, the acrosomal enzyme activity was detected by the BAEE/ADH method, the NOS activity was detected by the redoxreaction assay, and SOD level was measured with xanthine oxidase method. Compared with the control group, acrosomal enzyme activity of spermatozoa of the experimental group was significantly decreased (P < 0.01), NOS activity was apparently increased (P < 0.01), and the SOD level in seminal plasma was significantly decreased (P < 0.01). The infertility caused by positive AsAb in seminal plasma may be related to the changes in the acrosomal enzyme of spermatozoa and the SOD and NOS activities in seminal plasma.

The Change of Semen Superoxide Dismutase and Acrosin Activity in the Sterility of Male Patients with Positive Antisperm Antibody.

In this study, we report the change of semen superoxide dismutase (SOD) and acrosin activities in the male sterility patients with positive antisperm antibody (AsAb). The activity of SOD was measured by xanthine oxidase assay and sperm acrosin activity was calculated by BAEE/ADH. Our data show that compared with the normal fertility group, the semen SOD activity in AsAb-positive patients was significantly lower. Similarly, the sperm acrosin activity in AsAb-positive patients was also significantly lower. Our results suggest that the sterility resulting from positive AsAb may be related with the changes of semen SOD and sperm acrosin activities.

Effect of P/E-selectin blockage on antisperm antibody development and histopathological alterations in experimental orchitis.

AIM: This study aimed to evaluate the effect of P/E-selectin blockage on antisperm antibody (ASA) development and histopathological alterations in experimental orchitis. MATERIALS AND METHODS: Thirty-six Wistar albino-type male rats weighing 100-150 g were included in the study. Rats were allocated into six groups (n = 6) including control (CG), sham (SG), orchitis (OG), antimicrobial treatment (AG), P/E-selectin blockage (PESG), and both antimicrobial and P/E-selectin treatment (TG) groups. In CG, serum samples were taken from the tail vein prior to the procedure and followed by extraction of both testes. In SG, 1 ml of saline solution was injected in testicular parenchyma. OG was obtained by injecting 0.1 ml 106 cfu/ml Escherichia coli (0:6 strain) and 1 ml saline solution into the right testes. AG received ciprofloxacin (50 mg/kg/day) twice a day through gastrogavage 24 hours after generating orchitis. In PESG, P/E-selectin antibody (100 µg) was administered intravenously via the tail vein 24 hours after the induction of orchitis. Finally, both ciprofloxacin and P/E-selectin antibody were administered in TG 24 hours after the induction of orchitis for 14 days. At the end of treatment, 1 ml of serum sample was obtained to evaluate the ASA, P-selectin and E-selectin levels. In order to evaluate spermatogenesis (Johnsen score) and testicular injury (Cosentino score), both testes were extracted at the end of the 14th day. RESULTS: In orchitis-induced groups (OG, ATG, PSEG, TG), ASA levels were significantly increased at the 14th day when compared to SG (p < 0.05). In TG, ASA levels were decreased when compared to AG. However, similar alteration in ASA levels was not detected in PSEG (p > 0.05). In OG and AG, P-selectin levels were decreased at the 14th day when compared to levels observed on 0 day (p < 0.05). E-selectin levels on 0 day showed that each group had higher levels of E-selectin when compared to CG (p > 0.05). There was no significant difference regarding E-selectin when compared to CG (p > 0.05). No significant differences regarding E-selectin levels were detected on the 0th and 14th days between AG and CG (p > 0.05). When the Cosentino and Johnsen scores were compared among groups, TG and PSEG has decreased scores of Cosentino than OG on the right testicle (p < 0.05). In contrast, an increased Johnsen score was detected in TG and PSEG when compared to OG (p < 0/05). No significant difference was detected for both Cosentino and Johnsen scores on the left testicle (p > 0.05). There was no difference with regard to the right and left testicular injury in TG. In P/E-blocked groups, decreased histopathological alterations were observed in the contralateral testis. CONCLUSION: P/E-selectin blockage may reduce ASA production after orchitis when combined with antimicrobial treatment. P/E-selectin blockage not only has a protective effect on blood-testis barrier but also decreases the histopathological alterations in both the affected and contralateral testis. Histopathological parameters of spermatogenesis may also be prevented by P/E-selectin blockage in experimental orchitis.

Testicular microlithiasis is not a risk factor for the production of antisperm antibody in infertile males.

Testicular microlithiasis (TM) is a pathological event characterised by the presence of microliths within the testicular entities, and such calcium deposition is thought to have deleterious impacts on the structure of blood-testis barrier (BTB). Breaches in the BTB appear to be a risk factor for antisperm antibody (ASA) production, which is reported to have negative influence on human fertility. Thus, the theories are provocative that ASA formation is elicited in TM men, and the resultant ASA will accordingly affect the fecundity in these men. To illustrate these hypotheses, this study enrolled 22 infertile men incidentally diagnosed with TM by testicular ultrasound evaluation. Sperm samples were collected, and direct immunobead test was used to determine the ASA levels. None of the infertile men with TM were found to display significant levels of ASA, whilst relatively abnormal sperm parameters in these cases were revealed by semen analysis. These observations suggest that TM exposure does not increase the risk of ASA production in infertile men, and therefore, ASA is discarded as an active participant in the development of infertility in TM men. Nevertheless, disrupted spermatogenesis resulting from TM may, at least in part, have certain implications for the pathogenesis of TM-associated infertility.

Toward standardization of the cut-off value for the direct immunobead test using the postcoital test in immunologically infertile males.

PURPOSE: There is a need to improve our understanding of the cut-off value of the direct immunobead test (D-IBT). METHODS: The subjects were 26 D-IBT-positive and 140 D-IBT-negative males. The results of post coital tests (PCTs) for each subject were examined. RESULTS: A significant difference was observed in abnormal PCTs between values <20 % and those ≥20 % (P = 0.02). However, there was no significant difference in abnormal PCTs between values <50 % and those ≥50 % (P = 0.084). CONCLUSIONS: A cut-off value of 20 % was correlated with the possibility of conception on treatment with IUI. The D-IBT is a screening test, and the value of 20 % initially suggested by Bronson et al. seems to be more appropriate than that of 50 % in the criteria defined by the World Health Organization.

Analysis of the changes of antisperm antibody, antiendometrial antibody and anti-mulerian hormone in serum of women with infertility / 中国综合临床

ObjectiveTo examine the level of antisperm antibody (ASAb),antiendometrial antibody (EMAb) and anti-mulerian hormone(AMH) in serum of women with infertility and to provide a reliable basis for prediction,diagnosis and treatment of infertility.Methods Two hundred cases of women with infertility visiting our hospital between May.2010 and May.2011 were chosen as the infertility group and 100 cases of women with fertility experience visiting our hospital at the same period with the infertility group were chosen as the controlgroup.We examined the presence of ASAband EMAb and the levelsof AMHof the participants.ResultsIn the infertility group,the total positive rate of serum ASAb was 27.5％ (55/200) and the positive rates of ASAb-IgG,ASAb-IgM and ASAb-IgA were 11.5 ％ (23/200),22.5 ％ (45/200) and 9.5 ％(19/200) respectively.While in the control group,the rates were 6.0％ (6/100),1.0％ (1/100),0 and 2.0％(2/100) respectively ( x2 =5.33,5.37,5.41,4.05 ;P ＜ 0.05 ).The total positive rate of EMAb was 148.5％(97/200),and the positive rates of EMAb-IgG and EMAb-IgM were 13.5％ and 32.5％ respectively,and EMAb-IgG + EMAb-IgM positive rate was 5.5％ (11/20).These parameters were significantly higher than those in the control group [0,1.0％ (1/100),3.0％ (3/100),0,x2 =5.01,5.24,5.16,5.33 ;P ＜0.01 ].There was significantly difference on the level of AMH between the experimental group and the control group [ (5.39 ±1.42) μg/L vs.(2.55 ± 1.27 ) μg/L,t =5.39,P ＜ 0.01 ].Significant correlation was found between ASAb and EMAb( x2 =6.27,P =0.03) by correlation analysis.ConclusionThe level of AMH and the positive rates of ASAb and EMAb are higher in women with infertility than in normal people. Detection of ASAb,EMAb and AMH are important in in finding cause for infertility and taking appropriate measures to treat infertility

Frequency of antisperm antibodies in infertile women.

BACKGROUND: Infertility is one of the common problems seen in couples of reproductive age. Presence of antisperm antibodies in semen and serum are amongst the causes of immunoinfertility. This study was performed to determine antisperm antibodies in cervicovaginal secretions and serum of infertile women and also measure serum levels of immunoglobulins (IgG, IgM and IgA). METHODS: The study consisted of 45 infertile women consulting the Kammal El-Sammrari Hospital for infertility from 2008 to 2009 and the control group consisted of 30 fertile women. Serum levels of immunoglobulins (IgG, IgA and IgM) were measured in the participants using single radial immune diffusion. Antisperm antibodies (ASAs) were detected in the serum of both infertile and control groups using indirect immune fluorescence test. ASAs were also detected in cervicovaginal secretion using direct sperm agglutination test in both infertile and control groups. RESULTS: Antisperm antibodies were found in the cervicovaginal secretions (62.2%) and sera (64.4%) of infertile women which were significantly higher (p <0.001) than those of the control group (3.3% and 3.3% respectively). There was a significant increase (p <0.001) in serum levels of IgG and IgA of infertile women (16.2 and 3.25 g/L respectively) compared with the healthy control group (7 and 1.2 g/L). CONCLUSION: Humoral immune response and antisperm antibodies may contribute to reproductive failure in couples of reproductive age.