Antisperm antibodies (ASAs) are assumed to be a possible causative factor for male infertility, with ASAs detected in 5%-15% of infertile men but in only 1%-2% of fertile ones. It remains unclear whether ASAs have an adverse effect on the outcome of in vitro fertilization (IVF) or intracytoplasmic sperm injection (ICSI). This study investigated differences in the rates of fertilization, pregnancy, and live births associated with serum ASA-positive and ASA-negative men following IVF or ICSI. Five hundred and fifty-four consecutive infertile couples undergoing IVF (n = 399) or ICSI (n = 155) were included. The two-sample two-sided t-test and Chi-square or Fisher's exact test was used for statistical analysis. Lower rates of fertilization (41.7% vs 54.8%, P = 0.03), good embryos (18.9% vs 35.2%, P = 0.00), pregnancy (38.5% vs 59.4%, P = 0.00), and live births (25.8% vs 42.5%, P = 0.00) were observed in men of the IVF group with a positive serum ASA than in those with a negative ASA. ASA positivity/negativity correlated with pregnancy rates (P = 0.021, odds ratio [OR]: 0.630, 95% confidence interval [CI]: 0.425-0.932) and live birth rates (P = 0.010, OR: 1.409, 95% CI: 1.084-1.831) after controlling for the female serum follicle-stimulating hormone level and the couple's ages at IVF. Women coupled with ASA-positive men had lower live birth rates with IVF than with ICSI (25.8% and 47.4%, respectively; P = 0.07). Women coupled with ASA-positive men had lower rates of pregnancy and live births following IVF than those coupled with ASA-negative men but had a similar outcome with ICSI.
Adult ; Antibodies/pharmacology* ; Cohort Studies ; Female ; Fertilization ; Fertilization in Vitro/methods* ; Humans ; Infertility, Male/therapy* ; Live Birth ; Male ; Pregnancy ; Pregnancy Outcome ; Pregnancy Rate ; Sperm Injections, Intracytoplasmic/methods* ; Spermatozoa/immunology* ; Treatment Outcome ; Young Adult

Up to 15% of male infertility has an immunological origin, either due to repetitive infections or to autoimmune responses mainly affecting the epididymis, prostate, and testis. Clinical observations and epidemiological data clearly contradict the idea that the testis confers immune protection to the whole male genital tract. As a consequence, the epididymis, in which posttesticular spermatozoa mature and are stored, has raised some interest in recent years when it comes to its immune mechanisms. Indeed, sperm cells are produced at puberty, long after the establishment of self-tolerance, and they possess unique surface proteins that cannot be recognized as self. These are potential targets of the immune system, with the risk of inducing autoantibodies and consequently male infertility. Epididymal immunity is based on a finely tuned equilibrium between efficient immune responses to pathogens and strong tolerance to sperm cells. These processes rely on incompletely described molecules and cell types. This review compiles recent studies focusing on the immune cell types populating the epididymis, and proposes hypothetical models of the organization of epididymal immunity with a special emphasis on the immune response, while also discussing important aspects of the epididymal immune regulation such as tolerance and tumour control.
Adaptive Immunity ; Animals ; Epididymis/immunology* ; Fertility/immunology* ; Genital Neoplasms, Male/immunology* ; Humans ; Immunity, Innate ; Infertility, Male/immunology* ; Male ; Spermatozoa/immunology*

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). 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.
Antibodies, Bacterial ; analysis ; Humans ; Infertility, Male ; immunology ; microbiology ; Male ; Semen ; Sperm Count ; Spermatozoa ; immunology ; Ureaplasma Infections ; diagnosis ; immunology ; Ureaplasma urealyticum ; immunology

ObjectiveTo investigate the role of the serum inhibin B (INHB) level in evaluating the testicular function of the prepubertal patient with varicocele (VC) after high ligation of the spermatic vein (HLSV).

METHODSThis study included 31 prepubertal male patients with left VC, averaging 12.55 years of age and 9 complicated by right VC. We collected peripheral blood samples before and at 4, 12 and 26 weeks after HLSV as well as spermatic venous blood samples intraoperatively for determination of the levels of follicle-stimulating hormone (FSH), luteinizing hormone (LH), testosterone (T), anti-sperm antibody (AsAb) and serum INHB by ELISA.

RESULTSCompared with the baseline, statistically significant differences were observed in the INHB level in the peripheral blood at 12 and 26 weeks after operation (［255.18 ± 69.97］ vs ［141.78 ± 59.82］ pg/ml, P < 0.05) and that in the spermatic venous blood intraoperatively (［255.18 ± 69.97］ vs ［412.44 ± 259.42］ pg/ml, P < 0.01). Spearman's analysis showed a negative correlation between the level of INHB and that of FSH (r = －0.224, P < 0.01).

CONCLUSIONSThe level of serum INHB in the peripheral blood of the prepubertal VC patient is decreased within 6 months after HLSV and negatively correlated with that of FSH. The levels of INHB and FSH may well reflect the testicular function of the prepubertal VC patient.

Adolescent ; Antibodies ; blood ; Biomarkers ; blood ; Child ; Follicle Stimulating Hormone ; blood ; Humans ; Inhibins ; blood ; Luteinizing Hormone ; blood ; Male ; Spermatozoa ; immunology ; Testosterone ; blood ; Varicocele ; blood

C57BL/6N is the most widely used inbred mouse strain applied in a wide variety of research areas including cancer, cardiovascular biology, developmental biology, diabetes and obesity, genetics, immunology, neurobiology, and sensorineural research. To compare the fertilization rates of C57BL/6NKorl mice with two commercial C57BL/6N stocks, differences in reproductive organ structures, sperm and egg numbers, fertilization rates, and embryo development rates among C57BL/6NKorl (Korea FDA source), C57BL/6NA (USA source), and C57BL/6NB (Japan source) mice were determined. Among the stocks, no significant differences were detected in organ weight and histological structure of male and female reproductive organs, although body weight was higher in C57BL/6NKorl mice than that in the other groups. The concentration and morphology of sperm and eggs in C57BL/6NKorl mice were similar to those of C57BL/6NA and C57BL/6NB mice. Furthermore, the three stocks had similar in vitro fertilization and embryo development rates, although these rates tended to be higher in C57BL/6NB mice. Pup body weight was higher in C57BL/6NKorl and C57BL/6NB mice than that in C57BL/6NA mice. The results of the present study suggest that C57BL/6NKorl, C57BL/6NA, and C57BL/6NB mice obtained from three different sources have similar fertilization and embryo development rates, although there were slight differences in the magnitude of their responses rates.
Allergy and Immunology ; Animals ; Biology ; Body Weight ; Developmental Biology ; Eggs ; Embryonic Development ; Female ; Fertilization in Vitro ; Fertilization* ; Genetics ; Humans ; Male ; Mice* ; Mice, Inbred Strains ; Neurobiology ; Obesity ; Organ Size ; Ovum ; Pregnancy ; Spermatozoa

Objective: To prepare a polyclonal antibody against human lysozyme-like protein 4 (LYZL4) expressed in the prokaryotic system and identify the distribution of LYZL4 in the testis. METHODS: The full-length cDNA of LYZL4 was cloned into the pET32a plasmid and the expression of the recombinant LYZL4 (rLYZL4) was induced by IPTG. The rLYZL4 was purified by Ni-NTA and chitin affinity chromatography respectively and its bactericidal activity was observed by bilayer agar plate diffusion assay. The purified rLYZL4 was used as an immunogen to generate the polyclonal antibody, followed by examination of the antibody titer by ELISA and its specificity by Western blot. The distribution of LYZL4 in human tissue, sperm and seminal plasma was identified and its subcellular localization in the testis was determined by immunohistochemistry. RESULTS: rLYZL4 was expressed efficiently in the prokaryotic system and exhibited no bacteriolytic activity against M. lysodeikticus and E. coli. The anti-rLYZL4 polyclonal antibody could bind the recombinant protein with a high sensitivity and specificity. LYZL4 was identified in the testis, epididymis and sperm protein extracts and localized in the acrosomal region of round and elongating spermatids. CONCLUSIONS An anti-rLYZL4 polyclonal antibody was successfully prepared using the prokaryotic expression system. LYZL4 was detected in the acrosomal region of round and elongating spermatids, suggesting an association with the structure and function of the acrosome.
Acrosome ; immunology ; Animals ; Antibodies ; analysis ; Blotting, Western ; DNA, Complementary ; Enzyme-Linked Immunosorbent Assay ; Epididymis ; immunology ; Escherichia coli ; Humans ; Immunohistochemistry ; Male ; Muramidase ; genetics ; immunology ; Plasmids ; Recombinant Proteins ; genetics ; Semen ; immunology ; Spermatozoa ; immunology ; Testis ; immunology

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) and micro-TESE groups (2.50% vs 15.00%, P <0.05), while the serum T level markedly decreased in the two groups (［16.52 ± 6.25］ vs ［9.25 ± 5.76］ nmol/L and ［14.16 ± 5.45］ vs ［8.23 ± 4.12］ nmol/L, P <0.05); the levels of serum INHB were remarkably reduced at 1 and 3 months in the TESE (［70.56 ± 23.17］ vs ［42.63 ± 15.34］ and ［44.05 ± 18.47］ pg/ml, P <0.05), TESA (［68.71 ± 14.74］ vs ［40.55 ± 20.51］ and ［42.11 ± 19.34］ pg/ml, P <0.05), TEFNA (［76.81 ± 27.04］ vs ［46.31 ± 19.28］ and ［48.32 ± 20.54］ pg/ml, P <0.05), and micro-TESE groups (［74.74 ± 28.35］ vs ［45.27 ± 18.83］ and ［47.64 ± 28.34］ 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.
Antibodies ; blood ; Azoospermia ; blood ; physiopathology ; Follicle Stimulating Hormone ; blood ; Humans ; Inhibins ; blood ; Male ; Sperm Retrieval ; Spermatozoa ; immunology ; Testis ; metabolism ; physiopathology ; Testosterone ; blood

OBJECTIVETo prepare a specific polyclonal antibody against full-length SUN5 for detecting the expression of SUN5 in human germ cells.

METHODSBioinformatic methods were used to compare the full-length SUN5 and its variant SUN5β, and a short peptide was designed based on the differential region to prepare SUN5 antibody. The prepared antibody was used to detect the expression of SUN5 in Ntera-2 cells and in human germ cells by Western blotting and immunofluorescence assay.

RESULTSThe short peptide was correctly synthesized and SUN5 antibody was obtained and purified. Western blotting showed that the prepared antibody was capable of recognizing full-length SUN5 in Ntera-2 cells, and SUN5 expression was localized on the nuclear membrane and in the cytoplasm as shown by immunofluorescence assay. Using this antibody, we detected SUN5 expression in the spermatocytes, round spermatids and sperms in human germ cells.

CONCLUSIONWe successfully prepared SUN5-specific antibody. SUN5 is expressed in the spermatocytes, round spermatids and sperms in human germ cells, suggesting its important role in spermatogenesis.

Antibodies ; chemistry ; Blotting, Western ; Cytoplasm ; metabolism ; Fluorescent Antibody Technique ; Humans ; Male ; Nuclear Envelope ; metabolism ; Proteins ; immunology ; metabolism ; Spermatids ; metabolism ; Spermatocytes ; metabolism ; Spermatogenesis ; Spermatozoa ; metabolism

OBJECTIVETo prepare rabbit anti-mouse zona pellucida 2 (mZP2) polyclonal antibodies and test their immunoactivity.

METHODSRecombinant proteins of mZP2 expressed in Rosetta transformant was separated by SDS-PAGE, and the gel strips containing the recombinant mZP2 were cut out and emulsified to immunize New Zealand white rabbits. The antibody response of the antiserum was detected by ELISA, and the specificity of the antiserum was verified by immunohistochemical assay. The effect of the antiserum on the binding of oocytes with acrosomal reacted sperm was tested by sperm-egg binding assay.

RESULTSELISA results showed that the immunized rabbit produced anti-mZP2 antiserum. The antiserum reacted specifically with the zona pellucida of mouse ovarian sections. Sperm-egg binding assay showed that treatment of the oocytes with the anti-mZP2 antiserum caused decreased binding of zona pellucida with the acrosomal reacted sperm by 43.7%.

CONCLUSIONWe obtained rabbit anti-mouse ZP2 polyclonal antibodies that can inhibit the binding of oocytes with acrosomal reacted sperm.

Animals ; Antibodies ; immunology ; Antibody Formation ; Egg Proteins ; immunology ; Female ; Immune Sera ; Male ; Membrane Glycoproteins ; immunology ; Mice ; Oocytes ; Rabbits ; Receptors, Cell Surface ; immunology ; Recombinant Proteins ; immunology ; Sperm-Ovum Interactions ; Spermatozoa ; Zona Pellucida Glycoproteins

OBJECTIVETo detect the expressions of interleukin-6 (IL-6) and soluble intercellular adhesion molecule-1 (sI- CAM-1) in the seminal plasma of infertile men and explore the role of inflammatory cytokines in male immune infertility.

METHODSBased on the results of the sperm-bound antibody immunobead test, 123 males with clinically suspected infertility were divided into an immune infertility group (n = 41), other infertility group A (n = 37), and other infertility group B (n = 45). The immune infertility patients were further subdivided into a leukocyte-positive and a leukocyte-negative group according to the results of leukocyte peroxidase staining. The control group included 31 males confirmed to be fertilein the clinic. Statistical analyses were conducted on the differences in inflammatory cytokines expressions and main parameters in the seminal plasma among different groups. The seminal liquefaction time was measured by visual and microscopic observation, sperm concentration and motility detected using the computer-assisted sperm analysis system, sperm viability determined by hypotonic swelling assay, and the expression levels of IL-6 and sICAM-1 meas- ured by ELISA.

RESULTSThe infertility groups showed significantly lowers perm viability (P < 0.05) and progressive motility (P < 0.01) than the fertile control, but no remarkable differences from the latter in sperm concentration (P > 0.05) and semen liquefaction time (P > 0.05). No statistically significant differences were observed in seminal parameters between the immune infertility group and other infertility groups (P > 0.05). The IL-6 and sICAM-1 levels in the seminal plasma were extremely significantly higher in the im- mune infertility group ([37.92 ± 17.01] ng/L and [89.15 ± 41.82] ng/ml), other infertility group A ([22.23 ± 13.77] ng/L and [67.81 ± 33.24] ng/ml), and other infertility group B ([18.75 ± 14.32] ng/L and [53.25 ± 27.09] ng/ml) than in the normal control group ([9.47 ± 5.76] ng/L and [19.46 ± 9.77] ng/ml) (P <0.01) , with remarkable differences between the immune infertility group and the other two infertility groups (P < 0.05). The leukocyte-positive patients showed significantly increased levels of IL-6 ([49.25 ± 21.46] ng/L) and sICAM-1 ([104.36 ± 46.41] ng/ml) as compared with the leukocyte-negative ones ([31.38 ± 15.54] ng/Land [80.38 ± 35.52] ng/ml) (both P < 0.05).

CONCLUSIONIL-6 and sICAM-1 in the seminalplasma are involved in male immune infertility.

Biomarkers ; analysis ; Cytokines ; analysis ; Enzyme-Linked Immunosorbent Assay ; Humans ; Infertility, Male ; classification ; immunology ; Intercellular Adhesion Molecule-1 ; analysis ; Interleukin-6 ; analysis ; Male ; Semen ; chemistry ; immunology ; Semen Analysis ; Sperm Count ; Spermatozoa