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.

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.

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.

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.

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.

ACA、AsAb与不孕及反复自然流产的关系

目的探讨抗心磷脂抗体(ACA)、抗精子抗体(AsAb)与不孕、反复自然流产的关系。方法采用酶联免疫吸附法(ELISA)对124例不孕患者(不孕组)、182例反复自然流产(流产组)及68例无流产史且已分娩的健康妇女(对照组)血清ACA及AsAb进行测定分析并比较。结果不孕组ACA阳性率为45.16%,AsAb阳性率为33.06%,均显著高于对照组(P<0.01);流产组ACA总阳性率为48.90%,AsAb阳性率为26.92%,均显著高于对照组(P<0.01)。结论妇女不孕及反复自然流产与体内ACA和AsAb的存在有关。

益气活血汤对男性AsAb阳性病人外周血T细胞亚群分类的影响

目的 :通过观察中药益气活血汤对男性免疫性不育的作用 ,进一步研究中药对生殖免疫的作用机理。 方法 :30例血清抗精子抗体 (AsAb)阳性的不育病人 ,服用益气活血汤 2个月 ,测定用药前后外周血CD3,CD4 ,CD8以及CD4 CD8比值的变化。 结果 :治疗前后CD3数值变化不明显 (P >0 .0 5 ) ,治疗后CD4数值明显降低 (P <0 .0 5 ) ,CD8数值明显升高 (P <0 .0 5 ) ,CD4 CD8数值明显降低 (P <0 .0 5 )。 结论 :中药益气活血汤可能通过影响外周血T细胞亚群分类的水平 ,从而起到调节机体免疫力。

慢性前列腺炎/慢性盆腔疼痛综合征患者精浆成分和相应精子质量改变的研究进展

前列腺炎是前列腺三大常见疾病之一,此外分别是前列腺增生和前列腺癌。约50%的男性一生之中曾患前列腺炎。慢性前列腺炎/慢性盆腔疼痛综合征(CP/CPPS)患者中不育症的发生率显著高于正常患者,这主要与慢性前列腺炎患者精液成分的改变有关。本综述通过使用“慢性前列腺炎”、“慢性盆腔疼痛综合征”、“精子”、“精液”和“精浆”等关键词及术语,在PubMed和医学线检索了截至2024年之前发表的人类和动物研究的综述文章、临床试验和病例报告及相关引文。综合回顾了既往对于慢性前列腺炎与精浆变化及精子质量相关研究和调查,通过CP/CPPS患者前列腺液的成分的改变导致精浆成分发生变化的角度,讨论其对精子质量的影响。

男性不育者精浆C3d和C3的测定及与AsAb的关系

用ELISA法检测精浆中的C3d、C3含量，并计算C3d／C3的比值．结果表明，抗精子抗体（AsAb）阳性的不育病人精浆C3d含量及C3d／C3比值明显高于生育组，而AsAb阴性的不育病人糖浆C3d、C3及C3d／3的比值与生育组间则无明显差异。以C3d高于0．8mg／L为阳性，AsAb阴性和AsAb阳性的不育者C3d阳性率分别为19．2％和81．3％，二者差异非常显著（P＜0．01）。提示ASAb可能与补体活化有关，导致C3裂解增加，因此精浆C3d、C3的检测可反映精浆内补体激活的水平，可作为男性免疫性不育的一个重要指标．

SDS-PAGE分离不孕奶牛宫颈黏液AsAb

免疫不孕严重影响奶牛正常繁殖,相对于其他不孕类型不容易被确诊,准确对孕牛与不孕牛之间宫颈黏液AsAb的差异进行定性和定量分析,对奶牛免疫不孕的治疗研究有重要的意义。本试验将浅盘凝集检测和实际生产结果相结合确定的不孕奶牛和可孕奶牛作为研究对象,对宫颈黏液中蛋白进行SDS-PAGE电泳并对其分离条件进行优选。结果表明,本试验建立了一种在8%分离胶上,以15μL上样量,100V,电泳3~4h的SDS-PAGE分离奶牛宫颈黏液的有效方法;对浅盘凝集检测和实际生产记录相结合确定AsAb免疫性不孕奶牛的宫颈黏液,用SDS-PAGE分离得到一种分子量为22~25kDa蛋白,初步确定为AsAb轻链;发现100%(24/24)不孕奶牛的宫颈黏液中存在AsAb轻链,14.3%(2/14)可孕牛宫颈黏液中存在AsAb轻链,但表达量明显低于不孕牛。

CXL对AsAb阳性精液常规及精子膜完整性的影响

本实验观察了ＡｓＡｂ 阳性患者的精液常规分析（ＳＦＡ）的某些参数和肿胀精子率（评价精子膜的完整性） 的改变。并进一步观察了中药消抗安胎口服液（ＣＸＬ） 治疗ＡｓＡｂ 阳性患者的疗效。结果表明，ＡｓＡｂ 阳性患者精液ＳＦＡ 的精子密度、精子计数以及肿胀精子率较对照组为低，同时观察到，服用ＣＸＬ后，上述指标均得到明显改善。

不育相关AsAb的免疫印迹分析

用精子膜抗原Ｗｅｓｔｅｒｎｂｌｏｔ分析法，对７８例生育、１９８例不孕、２２例流产者及４４例中晚期妊娠孕妇血清抗精子抗体（ａｎｔｉｓｐｅｒｍａｎｔｉｂｏｄｉｅｓ，ＡｓＡｂ）进行研究。结果发现，抗分子量８５０００、５１０００、３００００精子抗原的ＡｓＡｂ与不孕相关；抗分子量８５０００、７９０００、６９０００，５１０００精子抗原的ＡｓＡｂ与女性不孕相关；不孕女性ＡｓＡｂ种类（在Ｗｅｓｔｅｒｎｂｌｏｔ上的反应条带数）明显多于生育女性。抗分子量８５０００、７９０００、５８０００精子抗原的ＡｓＡｂ与女性ＲＳＡ相关。本研究未发现不孕组男性、流产组男性及中晚期妊娠孕妇各种分子量精子抗原的ＡｓＡｂ检出率与生育组同性别相比有显著不同。所发现的这些ＡｓＡｂ可能与不育有密切关系，需要进一步研究和证实。

AsAb与男性不育的相关性研究

目的观察不育男性血清、精浆中 AsAb 与精液各指标之间的关系,揭示 AsAb 与不育症的相关性及致病机制。方法 ELISA 法测定127例男性不育患者及86例正常生育者血清、精浆中的 AsAb 以及精液的各项指标。结果不育者血清中的 AsAb 阳性率(29.9%)明显高于正常生育组(4.7%),p<0.05,不育患者血清、精浆 AsAb 水平变化能导致精子的活动力明显下降,呈显著性正相关(p<0.05)。结论AsAb 与男性不育有密切关系,可引起精子活动力明显下降,但对精子密度及液化无明显影响。

不孕症免疫因素AsAb,IFN-γ的相关性研究

目的 :观察不孕患者血清AsAb及其外周血细胞培养上清液中IFN γ的水平变化 ,了解AsAb与IFN γ的相关性 ,揭示AsAb ,IFN γ与不孕的关系及致病机理。方法 :酶联免疫吸附法 (EnzymelinkedImmunosorbentAssay ,ELISA)测定了 6 2例不孕患者及 5 0例正常生育者血清AsAb ,同时测定 5 0例不孕患者与 5 0例正常生育者外周血细胞培养上清液中γ 干扰素 (interferon γ ,IFN γ)水平。结果 :(1)不孕患者血清AsAb阳性率 (2 9.0 3% )显著高于正常生育组 (6 .0 0 % ,P <0 .0 5 ) ;(2 )外周血细胞培养上清液中IFN γ水平 ,不孕组为 (1.10 0± 0 .45 9) μg/L ,显著高于正常生育组 (0 .6 0 7± 0 .0 5 2 ) μg/L(P <0 .0 0 1) ;(3)不孕患者外周血细胞培养上清液中IFN γ水平 ,AsAb阳性组 (1.30 8± 0 .34 5 ) μg/L ,显著高于AsAb阴性组 (0 .836± 0 .2 6 5 ) μg/L(P <0 .0 0 1) ;(4)不孕患者血清AsAb水平与培养上清液中IFN γ水平呈显著正相关 (r=0 .812 6 ,P <0 .0 0 1)。结论 :(1)AsAb是与不孕有密切关系的重要免疫因素 ;(2 )不孕患者体内IFN γ水平明显高于正常生育组 ,提示患者体内细胞免疫功能状态异常增强 ,异常升高的IFN γ可能通过影响机体生殖免疫功能而导致不孕 ;(3)不孕患者体内异常高水平的IFN γ与AsAb?

反复自然流产患者血清P、AsAb、AcA和EmAb水平检测的临床意义

目的探讨反复自然流产患者血清P、AsAb、AcA和EmAb水平的变化及意义。方法应用放射免疫分析法和酶联法对32例反复自然流产患者进行血清P、AsAb、AcA和EmAb水平检测,并与35例正常妇女作比较。结果反复自然流产患者血清孕酮(P)水平非常显著的低于正常人组(P<0.01),AsAb阳性率为78.1%(25/32),AcA阳性率为37.5%(12/32),EmAb阳性率为43.8%(14/32),与正常人组比较差异有非常显著性(P<0.01)。结论血清P、AsAb、AcA和EmAb水平与反复自然流产有密切关系。

血清AsAb检测在女性不孕症中的诊断意义

目的探讨抗精子抗体(AsAb)检测在女性不孕症中的诊断意义。方法采用ELISA法检测485例患者血清中的AsAb;同时选用74例具有可比性的健康女性做正常对照。结果不孕组AsAb阳性率(31.7%,154/485)明显高于对照组(2.7%,2/74),且具有极显著性差异(P<0.01)。结论 AsAb与女性不孕症密切相关,进行AsAb检测对不孕症的诊断具有重要意义。

ELISA法定量测定抗精子抗体AsAb

目的 通过对抗精子总抗体（AsAb）的测定,探讨其与临床不育（或不孕）症的关系。方法 对107例志愿者AsAb的类型和含量用ELISA法进行了分型和定量研究。结果 总抗体的测定上限可达500U/ml;回收率在96.4%～103.0%之间;批内重复性为2.1%～5.0%;本试剂盒呈色稳定性较好,操作简单。通过测定其参考值范围为AsAb〈75.3U/ml。结论 该法准确可靠,可作为临床诊断不育或不孕症病因的辅助诊断方法 。

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.