Changes in soluble interleukin-2 receptor level in serum and Na^+ -K^+ -exchanging ATPase activity in semen of infertile men caused by antisperm antibody

Aim: To explore the possible mechanisms of male infertility caused by antisperm antibody (AsAb). Methods: Thesoluble interleukin-2 receptor (sIL-2R) level in serum was analyzed by ELISA and Na^+ -K^+ -exchanging ATPase activi-ty in semen by phosphorus (Pi) assay. Results: The slL-2R level in serum was significantly higher and the Na^+ -K^+ -exchanging ATPase activity in semen significantly lower in AsAb positive infertile men when compared with thecontrols. Conclusion: The AsAb titer varies with the slL-2R level in serum. A decrease in Na^+ -K^+ -exchangingATPase activity in semen may play a role in male infertility caused by AsAb.

Analysis of Sperm Membrane Protein Relevant to Antisperm Antibody by Two-Dimensional Gel Electrophoresis and Western Blotting

Objective To identify the sperm membrane proteins that are associated with antisperm antibody Methods Using antisperm antibody positive serum through unidimensional polyacrylamide gel electrophoresis and 2-dimensional gel electrophoresis followed by Western blot analysis to determine the molecular weights (MW) and isoelectric points (pI) of sperm membrane proteins that are associated with antisperm antibody. Results Eight kinds of MW with more than ten sperm membrane proteins can be recognized by antisperm antibody positive serum, of which the MWs and pI were 23 kD, 31 kD, 32 kD, 34 kD, 41 kD, 51 kD, 60 kD, 78 kD and 5.3, 5.5,5.7, 5.0, 5.3, 5.8, 6.0, 5.5～6.2, 4.6,5.1,5.5～5.8 respectively. The identification ratios of the sperm membrane proteins on 78 kD (60.7%), 60 kD (71.4%), 51 kD (14.9%) and 23 kD (14.29%) were higher. Conclusion The sperm membrane proteins with MW of 78 kD, 60 kD, 51 kD and 23 kD were associated with antisperm antibody and immunological infertility. Two- dimensional gel electrophoresis and Western blotting can precisely identify the sperm membrane proteins that are associated with antisperm antibody.

复发性流产患者抗精子抗体水平与T17/Treg及IL-10/IL-17关系

目的:分析复发性流产(RSA)患者抗精子抗体(AsAb)水平与辅助T型17细胞(Th17)/调节性T细胞(Treg)及白细胞介素(IL-10)/IL-17关系。方法:收集2022年1-12月于本院就诊的122例复发性流产患者为研究对象,53例女性RSA患者,69例配偶有RSA病史的男性患者;无不良孕产史已生育健康女性40例和配偶无RSA史男性55例作为对照组。检测纳入对象AsAb水平,Th17、Treg细胞数及IL-10、IL-17水平;采用Spearman相关性法分析AsAb阳性表达与Th17、Treg、IL-10、IL-17水平关系。结果:RSA组AsAb阳性8例,其中女性2例、男性6例,对照组阳性仅1例男性;RSA组AsAb阳性率(6.6%)高于对照组(1.1%);RSA组Th17(3.29±1.04个)、Th17/Treg(0.82±0.41)高于对照组(2.01±0.65个、0.29±0.46),Treg(4.02±1.34个)低于对照组(6.85±1.59个);IL-10(399.25±67.96 ng/ml)、IL-10/IL-17(1.34±0.25)低于对照组(602.09±54.31 ng/ml、2.96±0.38),IL-17(298.17±33.54 pg/ml)高于对照组(203.11±27.60 pg/ml);经Spearman法分析,AsAb阳性表达与Th17、Th17/Treg、IL-17水平呈正相关,与Treg、IL-10、IL-10/IL-17水平呈负相关(均P<0.05)。结论:RSA患者AsAb阳性率较高,且与Th17/Treg细胞及IL-10/IL-17相关。

EFFECT OF ELECTROACUPUNCTURE ON ANTISPERM ANTIBODIES IN MALE INFERTILITY PATIENTS

Aims: To explore the therapeutic effect of electroacupuncture (EA) for treatment of male immune infertility patients and to observe the effect of EA on antisperm antibody (AsAb) positive reaction. Methods: A total of 100 male infertility AsAb positive patients were randomized into EA group (n=50, BL 15, BL 17, 18, 23, etc.) and medication group (n=50, oral administration of prednisone, 5 mg/time, t.i.d.). Serum and sperm AsAb were determined with enzyme immunoassay technique. Results: Following 4 months’ treatment, the cure rates and the total effective rates of EA and medication groups were 40.4% (20 cases/50 cases) and 92.0% (45/50), 10.0% (5/50) and 64.0% (32/50) respectively. The cure rate of EA group was significantly superior to that of medication group (P<0.01). But, no significant difference was found between two groups in the total effective rate (P>0.05). After treatment, AsAb positive rates of both groups particularly that of EA group decreased significantly compared with pre treatment of each group (P<0.05-0.01). Conclusion: EA treatment can work well in the treatment of some immune mediated male infertility patients and possesses a favorable regulation action on AsAb reaction.

Antisperm Antibodies on the Surface of Spermatozoa before Ejaculation from Vasectomized Men

The antisperm antibodies (AsAbs) coated on spermatoZoa of the proximal vas deferens (sperm before ejaculation, SBE) from 48 fertile men who were volunteers of vasectomy and 24 vasectomized men who asked for vasovasotomy,were determined by immunobead test (IBT) and sperm-cervical mucus contact test (SCMC). The results showed that in fertile men there were no positive samples of SBE in IBT and SCMC. In vasectomized men positive samples of SBE were found in 79.4% for IgG, 38.2% for IgA and 35.5% for SCMC. The AsAbs on SBE could be found at the time of less than one year to more than 3 years after vasectomy. The AsAbs were still found on the semen samples at 1~3months after vasovasotomy. Our results also indicated that the incidence of AsAbs on SBE from vasectomized men could not predict the levels of AsAbs on their ejaculated sperm after vasovasotomy. There was no significant correlation between the levels of AsAbs in serium before vasovasotomy and those on SBE from vasectomized men.

Cross-neutralizing Anti-hemagglutinin Antibodies Isolated from Patients Infected with Avian Influenza A(H5N1) Virus

Objective To recover broad-neutralizing monoclonal antibodies(Bn Abs)from avian influenza A(H5N1)virus infection cases and investigate their genetic and functional features.Methods We screened the Abs repertoires of expanded B cells circulating in the peripheral blood of H5N1 patients.The genetic basis,biological functions,and epitopes of the obtained Bn Abs were assessed and modeled.Results Two Bn Abs,2-12 D5,and 3-37 G7.1,were respectively obtained from two human H5N1 cases on days 12 and 21 after disease onset.Both Abs demonstrated cross-neutralizing and Ab-dependent cellular cytotoxicity(ADCC)activity.Albeit derived from distinct Ab lineages,i.e.,V^H1-69-D2-15-JH^4(2-12D5)and V^H1-2-D3-9-JH^5(3-32 G7.1),the Bn Abs were directed toward CR6261-like epitopes in the HA stem,and HA2 I45 in the hydrophobic pocket was the critical residue for their binding.Signature motifs for binding with the HA stem,namely,IFY in VH1-69-encoded Abs and LXYFXW in D3-9-encoded Abs,were also observed in 2-12D5 and 3-32 G7.1,respectively.Conclusions Cross-reactive B cells of different germline origins could be activated and re-circulated by avian influenza virus.The HA stem epitopes targeted by the Bn Abs,and the two Ab-encoding genes usage implied the VH1-69 and D3-9 are the ideal candidates triggered by influenza virus for vaccine development.

Effects of autoantibodies against AT1-receptor and angiotensin Ⅱ on refractory hypertension

Objective The study will explore effects of the autoantibodies against AT1 receptor and angiotensin Ⅱ on the refractory hypertension. Methods Seventy-seven patients (46 men and 31 women) with essential hypertension were divided into groups of refractory hypertension (RH) and hypertension (HT) according to the 1999 WHO-ISH Guidelines for the Management of Hypertension. Forty normotensives (22 men) were recruited as controls. The mean age was 54. 3±13 years old in RH group, 53. 5±9 years old in HT group and 51. 2±11. 9 years old in normotensives (NT) group. The mean blood pressure was 154. 2±9. 4/98. 4± 8. 2 mmHg in RH group and 130. 1±7. 6/80. 5±6. 7 mmHg in HT group after combination drug therapy of hypertension for 4 weeks. Blood pressure in NT group was 120. 8±11. 7/76. 4 ± 7. 2 mmHg. The epitope of the 2nd extracellular loops of AT1 receptor was synthesized and used as antigens to screen the autoantibodies by ELISA. Plasma angiotensin (Ang) II were examined by a radioimmunoassay. Results The autoantibodies against AT1 receptor were positive in 18 (46. 15 %) patients with RH, in 4 (10. 5 % ) hypertension and in 3 (7. 5 % ) normotensives, P < 0. 01. Ang Ⅱwas 57. 01±52. 63 pmol/L in patients with RH. Both the autoantibodies positive and the Ang Ⅱ increasing were 4 (10. 3 % ) cases, both normal were 7 (17. 9 % ) cases, the autoantibodies positive or Ang II increasing was all of 14 (35. 9 % ) cases (x2 = 0. 09, P>0. 05) . There was no relationship between the autoantibodies against AT1 receptor and the angiotensin Ⅱ in refractory hypertension. Conclusion The autoantibodies against AT1 receptor and Ang Ⅱ might be two independent factors in developing of refractory hypertension. The findings suggest that AT1 receptor an-tagnist used in the treatment of refractory hypertension might have an important value.

INHIBITION OF ZYGOTE FORMATION AND EMBRYO DEVELOPMENT IN MICE IMMUNIZED WITH MONOCLONAL ANTISPERM ANTIBODIES

Passive immunization of female mice by the intraperitoneal administration of ascitic fluidscontaining monoclonal antisperm antibodies blocked the development of mouse zygotes tomorulae and blastocysts when cultured in vitro.Radioiodinated YWK-I monoclonal anti-bodies were prepared and administered intraperitoneally to Jemale mice.Significant radioac-tivity was found in washes of the oviductal lumens.The results suggest that antispermantibodies block fertilization and may have a deleterious effect on early developing embryos.

Effect of GABA on sperm acrosome reation in antisperm antibody positive patients

To investigate the effect of gamma-aminobutyric acid (GABA) on the rate of sperm acrosome reaction both in normal and antisperm antibody (AsAb) positive men. Methods: The sperm acrosome reaction was tested with triplestain technique in two groups of 18 men each. Results: (1) GABA increased the rate of sperm acrosome reaction both in normal and AsAb positive subjects (P<0.01); (2) GABA increased the Na^+-K^+-ATPase activity of sperm (P<0.01); (3)GABA increased the Ca^(2+)-ATPase activity of sperm (P<0.05); (4) GABA decreased the production of MDA and oxygen free radicals of sperm. Conclusion: GABA could regulate the rate of sperm acrosome reaction. (Chin J Andro12002; 16: 355)

Studies on relationship between semen viscosity and other semen parameters, Ureaplasma urealyticum infection and seminal plasma antisperm antibody

Objective: To study the relationship between semen viscosity and other semen parameters, Ureaplasma urealyticum (UU) infection and seminal plasma antisperm antibody (AsAb) in male infertiles. Methods: Semen parameters, Ureaplasma urealyticum (UU) infection and antisperm antibody (AsAb) were measured and analyzed in 4337 infertile men. Results: The seminal viscosity was higherr than normal in 65.02 % of 4337 male infertiles. The sperm motility and grade (a, b) motile sperm were significantly lower in the high viscosity group than in the normal viscosity group (P<0.05-0.01). The rate of abnormal morphology sperm was higher and duration of semen liquefaction was longer in the high viscosity than in the normal viscosity group (P<0.01). The seminal volume, sperm concentration and semen pH were not significantly different between the two groups. The semen viscosity is significantly higher in subjects with higher seminal WBC (>5/ HP) than in those with lower WBC (<5/HP). The positive AsAb and UU infection rates were significantly higher in the high viscosity group (P<0.01). Conclusion: The semen viscosity is related to other seminal parameters, as well as to UU infection and seminal AsAb.

Analysis of sperm membrane antigens relevant to antisperm antibody using Western blot

To identify the sperm membrane antigens associated with antisperm antibody. Methods: The antisperm antibody in serum was tested by ELISA. Antisperm antibody positive sera from 18 infertile men and 15 infertile women were used. The molecular weight (MW) of sperm membrane antigens associated with the antisperm antibody was analyzed with antisperm antibody positive serum using Western blot. Results: Eight kinds of MW of sperm membrane antigens were identified. The ratio of identification on the 78 KD(60.7 %), 60KD (71.4 %), 51 KD (14.9 %) and 23 KD (14.29 %) sperm antigen was higher than others. Conclusion: Sperm membrane antigens with MW of 78 KD, 60 KD, 51 KD and 23 KD were associated with antisperm antibody and immunological infertility. (Chin J Andro12002; 16: 345)

Determinantion and evaluation of antisperm antibodies and seminal immunosuppressive material in recurrent aborting couples

Circulating antisperm antibodies (AsAb) and immunosuppressive material in seminal plasma (SPIM)were determined by solid-phase enzyme staining assay and anticomplement test respectively in 686 patients with abortion (including 285 couples) . 241 fertile couple served as control. It’s found that the positive rate of AsAb in infertile patients was significantly higher than that in fertile control,being 36.6% vs 3.3% (P【0.001). AsAb was even more offen detected in recurrent aborting patients. Male patients whose spouses aborted 2-6 fetuses had significantly less SPIM than control, sperm count and sperm motility were also significantly decreased. But the incidence of pyospermia was significantly greater than that in control. It is concluded that AsAb and SPIM have played an important role in the development of recurrent abortions.

Anti-Sperm Antibodies: Prevalence of Ig A and Ig G Positive Serology among Infertile Patients in Kisangani, Democratic Republic of Congo

Introduction: Infertility in a couple is a stressful situation. Among the causes is the presence of antisperm antibodies that play a harmful role in reducing fertility and negatively influencing in vitro fertilization. The aim of this study is to determine their prevalence among infertile patients in Kisangani. Patients and methods: We conducted an analytical cross-sectional study in health facilities in the city of Kisangani. Of the 1379 patients who consulted for conception, 311 were tested for antisperm antibodies by ELISA. Results: Of the 311 patients, 121 tested positive (38.91%). Women were significantly more likely to be positive (p-value = 0.013). Ig G was positive in 32.48% of patients and Ig A in 10.93%. Age was a significant predictor of the number of positive patients (p-value < 0.001). Compared to patients of ≤30 years of age, having 31 to 40 years of age or 41 years of age and older increased respectively 2 and 3 times the risk of being positive. In secondary infertility, women are twice as likely to be positive. Conclusion: The prevalence of antisperm antibodies is very high among infertile patients in Kisangani with predominance among women. Older age increases the risk of positivity. This test should therefore be integrated into infertility investigations as often as possible.

Etiology of Antisperm Antibodies in the Serum of Virgins

Background: The study of ASA etiology is very important in the diagnosis and treatment of infertility. Studies of presence of antisperm antibodies in the bodies of unmarried women are very rare, so that this article aims at studying the possible causes and interpretations behind the development of antisperm antibodies in virgins. Methodology: The study included 5 single women with positive ASA. Description and clinical history of the patients was assessed by special questionnaire provided for this purpose. All laboratory investigations and diagnostic procedures were done in the hospital from 1st August to 15th December 2017. Results: The mean serum antisperm antibody concentrations (64.3 IU/ml) which is considered positive titer. The incidence of ASA among virgins is 2.22%. Complete blood count is normal except for slight increase in WBC count and percentage of basophils, monocytes and lymphocytes. This study recorded high concentrations of serum total IgG and IgM levels (1875 IU/ml and 295 IU/ml respectively). The UTI was confirmed by counting total bacterial concentration (178,250 CFU/ml) in the urine, and diagnosis of suspected causes showed the following species: Escherichia coli, Klebsiella pneumonia, Staphylococcus aureus, and Proteus mirabilis. Conclusions: There are two suggested mechanisms to explain ASA in virgins: 1) antigen cross-reactivity between sperm and bacterial antigens to which antibodies can react;2) induction of the immune system by antigens of sperm ingested into the gastrointestinal tract with contaminated food and drink. Recommendations: it is recommended to conduct a research study that include a large number of virgins for investigating ASA to confirm our results and build a scientific generalizations , in addition to animal studies for testing the role of sperm ingestion in the induction of immune system.

The Presence of Antisperm Antibodies in Semen of Subfertile Men—Positive ASA in Subfertile Men

Background: In more than 50% of male subfertility, the aetiology remains unknown. Antisperm antibodies (ASA) might be involved, however the exact role of ASA in unexplained male subfertility is not clear, yet. Objective: The aim of this study was to examine 1) the prevalence of ASA in subfertile men, 2) the possible causes of the presence of ASA, and 3) the influence of ASA on sperm parameters and fertilization including assisted reproductive technologies (ART) and pregnancy outcomes. Study Design, Size, Duration: In this retrospective single center study, all men with semen analyses between January 2003 and December 2017 were included as well as all subfertile couples getting treatment if at least one sperm analysis showed a spermMar test ≥50%. Methods: Collected parameters were: intoxications, medication and professions that could have an adverse effect on fertility, sperm parameters, the type and amount of ART, mode of conception, pregnancy rates and outcomes. Results: 3098 semen analyses were performed. In total, 233 ASA positive men were observed, including 175 subfertile couples with an ASA positive man in the additional analyses. The prevalence of ASA in the subfertile population was 8.2%. The presence of ASA was significantly associated with the presence of oligoasthenoteratozoospermia (OAT), asthenoteratozoospermia and asthenozoospermia (p = 0.008, p = 0.004, and p = 0.02 respectively). However, 50% of the couples with an ASA positive man became pregnant without ART. Conclusions: The presence of ASA did not seem to have a negative effect on spontaneously pregnancy rates or pregnancy rates after ART. Therefore, it might be justified to advice 6 - 12 months expectant management, before starting ART in ASA positive men.

Anti-Sperm Antibodies: Risk Factors of Positive Serology among Infertile Men Patients in Kisangani-Democratic Republic of Congo

Introduction: Antisperm antibodies (ASA) prevalence is high in Kisangani. ASA are one of the male factors of infertility that can reduce spermatozoa motility and cervical penetration or prevent acrosomal reaction and even alter embryo development. This study aims to determine the risk factors of ASA positivity among infertile men in Kisangani. Patient and Method: We conducted a case-control study during a period of 2 years in Kisangani on 111 men who consulted for conception and tested for ASA by ELISA. We did a simple pairing that concerned only the type of infertility. For a better balance between the number of cases and controls per stratum to provide better accuracy in the adjusted OR estimate, we performed a 1:1 match. Results: This study showed that businessmen [ORa = 5.0000(1.2452 - 20.0767)] and jobless [ORa = 5.8125(1.1477 - 29.4367)] were at risk of being positive for ASA. Violent blow to testicles [ORa = 6.7391(1.3455 - 33.7545)], cure of hernia [ORa = 4.3478(1.0722 - 17.6299)], Chlamydia infection [ORa = 4.7125(1.3405 - 16.5665)], leucospermia [ORa = 4.1429(1.0406 - 19.7155)] and presence of Staphylococcus aureus [ORa = 3.4444(0.6410 - 18.5082)] were associated to positive ASA. Conclusion: This study shows that factors that may lead to risky sexual behavior and physical or infectious trauma are important risk factors for ASA positivity. It is therefore necessary to search for them systematically during male infertility in order to guide the search for ASA.

Analysis of Antisperm Antibody in Cervix Mucus for Azoospermic Men’s Wives

ELISA method was used to detect antisperm antibodies in cervix mucus. Out of 400 women whose husbands had sperms, 92 were antisperm antibody positive (23% ). 42 of 200 women with azoospermic partners were antisperm antibody positive (21% ). No difference was found between the two groups. Hence, antisperm antibodies in azoospermic men's wives may be caused by sperm'sadhering antigens in semina.

Clinical Observation on Effect of Combination of Zhuanyindan (转阴丹) and Hormone in Treating MaleInfertility with Positive Antisperm Antibody

Objective:To explore the therapeutic effect of combination of Zhuanyindan (ZYD, 转阴丹, a self-made Chinese herbal preparation) and hormone in treating male infertility with positive antisperm antibody and its influence on nitric oxide (NO) level. Methods:Eighty-two patients were randomly divided (according to the digital list) into the WM group (n = 20, treated with prednisone), the TCM group (n=28, treated with ZYD) and the ICWM group (n=34, treated with prednisone plus ZYD). The clinical effect, negative converting rate of antisperm antibody, changes of NO level in semen and various parameters of sperm motion before and after treatment were observed. Results: The total effective rate in the ICWM group was 88.2%, that in theTCM group 75. 0% and in the WM group 65. 0%. Significant difference was seen in the ICWM and TCM group before and after treatment in NO level, sperm motion parameters, including linear motion speed, linearity, propulsion, whip frequency, sperm vitality and mean moving angle, and quality of semen ( P < 0. 05 or P < 0. 01). In the WM group, significant difference in comparison before and after treatment was seen in NO level, propulsion, whip frequency, mean moving angle and quality of semen, including vitality and survival rate (P<0. 01). Conclusion: Combination of Chinese herbs and hormone could lower the NO level in semen and improve the quality of sperm.Original article on CJITWM (Chin) 2004 ;23 (3): 223

Establishment of the Method of Immunohistochemistry Assay for the Detection of Scrapie in Chinese Short-Tailed Han Sheep by Monoclonal Antibody

The method of immunohistochemistry assay for the detection of scrapie in Chinese Short-tailed Han sheep was established using monoclonal antibody. Genomic DNA was isolated from Chinese Short-tailed Hart sheep blood. Using the polymerase chain reaction technique, PrP27-30 gene sequence was amplified from Chinese Short-tailed Han sheep genomic DNA. By recombinant DNA technology, the recombinant protein of Chinese Short-tailed Han sheep PrP27-30 was obtained. Then, using standard methodology of myeloma cell fusion, a panel of monoclonal antibodies was generated. With mAbs, scrapie in Chinese Short-tailed Han sheep was detected by immunohistochemistry assay. The recombinant protein of Chinese Short-tailed Han sheep PrP27-30 was obtained and a panel of six hybridoma cell lines secreting specific antibodies to Chinese Short-tailed Han sheep PrP27-30 related to scrapie was obtained with one fusion between myeloma Sp2/0 and spleen ceils from mice immunized with the purified recombinant protein. Four hybridoma cell lines can be used in immunohistochemistry assay for the detection of scrapie in Chinese Short-tailed Han sheep. So that the special monoclonal antibody developed in author＇s institute can be used to detect PrP^sc of scrapie in Chinese Short-tailed Han sheep by immunohistochemistry in China.

γ-氨基丁酸对精子顶体酶活性的影响

目的 :研究γ 氨基丁酸 (GABA)对生育男性精子及抗精子抗体 (AsAb)阳性病人的精子顶体酶活性的影响。 方法 :用BAEE ADH联合法同时测定GABA对精子顶体酶活性的影响。 结果 :GABA可明显提高正常及AsAb阳性精子顶体酶的活性 (P <0 .0 1) ;并使精子的Na+ K+ ATPase、Ca2 + ATPase、超氧化物歧化酶 (SOD)的活性增加(P <0 .0 1,P <0 .0 5 ,P <0 .0 0 1)。 结论