Anti-sperm antibodies from infertile patients and their cognate sperm antigens: a review. Identity between SAGA-1, the H6-3C4 antigen, and CD52

PROBLEM: The correlation of anti-sperm antibodies (ASA) with some instances of unexplained infertility implicates a role for these antibodies in blocking fertilization. Improved diagnosis and treatment of immunologic infertility, as well as a more complete understanding of the mechanism behind this phenomenon, are dependent on the identification and characterization of relevant sperm antigens. METHOD OF STUDY: In this article, we review literature on methods employed to identify sperm antigens using anti-sperm polyclonal and monoclonal antibodies from infertile patients and vasectomized men. Particular focus is given to approaches using human and mouse monoclonal antibodies to define the SAGA-1 human sperm antigen. RESULTS: ASA present in sera and genital tract secretions from infertile patients and vasectomized men have been employed in a variety of methods to identify sperm antigens. In an alternate approach, a monoclonal antibody (mAb), H6-3C4, was immortalized from the lymphocytes of an infertile woman who exhibited sperm-immobilizing titers. Subsequently, the sperm-agglutinating, murine S19 mAb was shown to react with the H6-3C4 cognate antigen. The H6-3C4 S19 cognate antigen, designated Sperm Agglutination Antigen-1 (SAGA-1), was characterized as a polymorphic, highly acidic, GPI-anchored glycoprotein on the surface of human spermatozoa. Purification with the S19 mAb followed by microsequencing demonstrated that the SAGA-1 core peptide is identical to CD52, a glycoprotein on the surface of human lymphocytes. Immunoblot analysis demonstrated that these two glycoproteins differed in carbohydrate composition. Thus, sperm SAGA-1 and lymphocyte CD52 represent glycoforms, glycoproteins with the same core peptide but with different carbohydrate structures. CONCLUSIONS: Autoimmunity to the SAGA-1 and/or CD52 glycoforms may lead to infertility. Structural and immunologic differences between these glycoproteins may be important factors in the etiology of immunologic infertility and other autoimmune disorders.     

178例男性不育患者Y染色体AZF微缺失筛查

目的探讨男性不育症患者Y染色体微缺失分子检测临床意义。方法应用多重PCR对178例不育症患者进行Y染色体AZFa、AZFb和AZFe基因微缺失检测。结果41例特发性无精症患者中有10例缺失,占24．3％;34例严重少精子症患者中有4例缺失,占11．7％;其余103例少弱精子症患者中没有检出缺失。结论在男性不育症患者中,Y染色体AZF基因微缺失是特发性无精子或严重少精子症发生的重要原因,基因检测可为正确诊断和合理治疗提供科学依据。     

育龄妇女不育不孕的实验室诊断及实验结果分析

目的探讨育龄妇女自身免疫抗体及生殖道衣原体(CT)、支原体(UU)感染与不育不孕患者的关系。方法选择2007年来我院门诊就诊和妇科住院的自然流产患者、不孕患者,没有反复流产史,且有一个健康婴儿的健康体检者,分别检测封闭抗体(APLA)、抗精子抗体(AsAb)、抗子宫内膜抗体(EMAb)、抗心磷脂抗体(ACA)解脲支原体(UU)和沙眼衣原体(CT)。结果自然流产患者中APLA阴性占75%,不孕患者APLA阴性占17.73%,对照组APLA阴性占8.14%;流产组、不孕组的APLA、AsAb、EMAb、ACA、UU、CT阳性检出率明显高于对照组。结论育龄妇女体内存在APLA、AsAb、ACA、EMAb及生殖道支原体或衣原体感染与不育不孕患者密切相关,对有不良孕产史及不孕的患者进行自身免疫抗体及生殖道衣原体、支原体的检测,可为诊断及治疗提供科学的依据。     

369例男性不育症患者的染色体核型分析

目的探讨男性不育与染色体核型异常的关系。方法按照精液常规检查结果,将369例男性不育症患者分为4组(无精子症组61例,少精子症组98例,弱精子症组117例,精液正常组93例),采用常规外周血淋巴细胞染色体制备方法对上述患者进行细胞遗传学检测。结果染色体异常检出率为8.4%(31/369),其中无精子症组19.7%(12/61),少精子症组7.1%(7/98),弱精子症组6.0%(7/117),精液正常组5.4%(5/93)。结论染色体异常是男性不育的主要遗传因素。     

男性不育患者Y染色体微缺失的研究

目的研究无精子症和少精子症患者与Y染色体位点缺失的相关性，建立Y染色体微缺失的分子诊断方法。方法采用多重PCR技术对53例染色体核型正常的无精子症和少精子症患者以及5例正常男性的无精子因子（azoospermia factor，AZF）区域的6个STS位点进行检测。结果5例精液正常男性未检出Y染色体微缺失；53例患者中6例有AZF区域的微缺失，总缺失率为11．3％。结论Y染色体微缺失是严重生精障碍的重要原因之一，无精子因子（AZF）候选基因在精子发生过程中可能起重要作用。     

Co-Expression of Mannose-Ligand and Non-Nuclear Progesterone Receptors on Motile Human Sperm Identifies an Acrosome-Reaction Inducible Subpopulation

PROBLEM : To determine whether surface expression of receptors for progesterone and mannose can be used to identify spermatozoa likely to undergo an acrosome reaction after zona binding and to compare the reactivity of these receptors with naturally occurring sperm head-directed anti-sperm antibodies (ASAs). METHOD : Progesterone binding sites on the surface of fresh and capacitated motile human sperm in relation to acrosome status were visualized using a cell-impermeant progesterone. Free progesterone and/or mannose ligands were compared for percent sperm binding and ability to induce an acrosome reaction. Western blots of sperm proteins localized to the plasma membrane and surface proteins precipitated following passive transfer of serum ASAs were probed with progesterone-horseradish peroxidase. The effects of the same ASAs on ligand binding and on the induced acrosome reaction were examined. RESULTS : The two receptors are located in close proximity on a subset of capacitated motile sperm and are coordinately cleared from the plasma membrane overlying the acrosomal cap prior to exocytosis. The surface appearance of functional binding sites for each ligand, however, is regulated by different mechanisms and the progesterone receptor alone is specifically precipitated by ASAs. Passive transfer of ASAs to capacitated sperm selectively inhibits the progesterone-stimulated acrosome reaction but not the ionomycin-induced acrosome reaction or the ability of sperm to bind mannose ligands. CONCLUSIONS : Sperm from fertile donors incubated under capacitating conditions in vitro can be subdivided into acrosome reaction inducible and noninducible subpopulations on the basis of the co-expression or total absence of these receptors. The combined data indicate that reaction of sperm surface progesterone receptors with ASAs contributes to the acrosome reaction insufficiency observed in anti-sperm immune infertility.     

张家口地区154例男性不育患者染色体分析

目的研究男性染色体异常与男性不育之间的关系。方法对154名患者的外周血淋巴细胞进行常规培养，然后作G带核型分析。结果在154名不育患者中，检出核型异常29例，其中有26例为数目异常（包括5例46，XY／47，XXY嵌合体），3例为结构异常。另外本实验还发现了2例男性假两性畸形患者。结论染色体异常是男性不育的重要原因之一。     

135例男性不育症的细胞遗传学分析

目的研究男性染色体异常与男性不育症的关系。方法用常规方法制备外周血淋巴细胞标本,对135例男性不育症患者进行染色体G显带核型分析。结果检出异常核型74例,占全部被检者55％。其中性染色体异常40例。占全部被检者30％,占异常核型54％;常染色体异常34例,占全部被检者25％,占异常核型46％。结论对男性不育症的患者进行染色体检查能对不育夫妇的诊断和治疗提供指导性依据。     

Molecular mimicry: An explanation for autoimmune diseases and infertility

Microorganisms execute an enthralling range of adjustments to survive in the host. Among the various strategies employed by microorganisms to surmount the host immune response, the phenomenon of molecular mimicry empowers the microorganisms to manoeuvre host physiology and cellular functions for their own advantage by mimicking the host proteins and initiating autoimmunity. This phenomena, by and large, has been studied in context of autoimmune diseases; however, its implications have also been reported in infertility. Hence, in this article, we provide a review of the various instances of molecular mimicry initiated by bacteria, parasites and viruses in the world of autoimmune diseases and infertility.     

Modalities for treatment of antisperm antibody mediated infertility: novel perspectives

Immunoinfertility because of antisperm antibodies (ASA) is an important cause of infertility in humans. The incidence of ASA in infertile couples is 9-36% depending on the reporting center. Early claims regarding the incidence and involvement of ASA in involuntary infertility were probably overemphasized, which has resulted in subsequent confusion, doubt, and underestimation of their clinical significance. No immunoglobulin that binds to sperm should be called an antisperm antibody in a strict sense unless it is directed against a sperm antigen that plays a role in fertilization and fertility. ASA directed against the fertilization-related antigens are more relevant to infertility than the immunoglobulins that bind to sperm associated antigens. Several methods have been reported for treatment of immunoinfertility. These include: immunosuppressive therapies using corticosteroids or cyclosporine; assisted reproductive technologies such as intrauterine insemination, gamete intrafallopian transfer, in vitro fertilization, and intracytoplasmic sperm injection; laboratory techniques such as sperm washing, immunomagnetic sperm separation, proteolytic enzyme treatment, and use of immunobeads. Most of the available techniques have side effects, are invasive and expensive, have low efficacy, or provide conflicting results. Recent findings using defined sperm antigens that have a role in fertilization/fertility have provided animal models and innovative novel perspectives for studying the mechanism of immunoinfertility and possible modalities for treatment. The better understanding of local immunity and latest advances in hybridoma and recombinant technologies, proteomics and genomics leading to characterization of sperm antigens relevant to fertility will help to clarify the controversy and to establish the significance of ASA in infertility.     

The prevalence of coeliac disease in infertility.

An increased incidence of reproductive problems, including infertility, miscarriage, low birth weight newborns, and shorter duration of breast-feeding, are known to exist in women with coeliac disease; some of these conditions are improved by a gluten-free diet. We have tried to ascertain the prevalence of coeliac disease in 99 couples who were being evaluated for infertility, compared with the known prevalence of silent disease in the population of Northern Sardinia, in which it is endemic. Of all women, four tested positive for at least two out of three markers: immunoglobulin A (IgA) antigliadin, immunoglobulin (IgG) antigliadin, and anti-endomysium antibodies, and underwent a jejunal biopsy; three had histological evidence of coeliac disease. One male partner was positive for two markers, and had a diagnostic jejunal biopsy. The prevalence of coeliac disease in infertile women seems higher (three out of 99, 3. 03%) in the study group than in the general population (17 out of 1607, 1.06%), and particularly in the subgroup with unexplained infertility (two out of 25, 8%, P < 0.03). Screening for coeliac disease should be part of the diagnostic work-up of infertile women, particularly when no apparent cause can be ascertained after standard evaluation.     

Cyclic corticosteroid immunosuppression is unsuccessful in the treatment of sperm antibody-related male infertility: a controlled study.

In this double blind cross-over study, 20 infertile men, who had sperm antibodies detected by the mixed antiglobulin reaction (MAR test) in the ejaculate and by the tray agglutination test (TAT) in serum, were treated with 40 mg/day prednisolone or placebo from days 1 to 10 of the partners' menstrual cycle. Patients were randomly allocated to different treatment groups. While group 1 started with placebo followed by verum for three consecutive cycles, group 2 began with verum and continued with placebo. All patients had regular intercourse (n = 19) or intra-cervical insemination at ovulation (n = 1). A post-coital test or a sperm penetration test was performed during verum and placebo regimes. Blood samples were drawn from the male partner at this time to control the efficacy of prednisolone treatment by checking the TAT titre. No pregnancy occurred during prednisolone or placebo treatment. In eight of 12 patients, post-coital testing showed little improvement and antibody titres decreased in seven of 16 patients. Side-effects from medication were reported by eight patients (seven verum and one placebo cycle) and caused treatment to be discontinued in two cases. Five patients' partners conceived at a later stage by intrauterine insemination with spermatozoa prepared by 'swim up' (n = 3) or by in-vitro fertilization (IVF n = 2). Thus high dose corticosteroid therapy was ineffective in achieving pregnancies induced by infertile men positive for antisperm antibodies. Since side-effects of corticosteroids should not be underestimated in otherwise healthy men, other reproductive techniques such as intrauterine insemination or IVF should be offered to such couples.     

Risk factors for female and male infertility: results of a case-control study.

In order to evaluate male and female risk factors for infertility in a case-control study, we have compared all couples in a French administrative region consulting for primary or secondary infertility (of more than one year's duration) with couples in which the woman had given birth during the year of the study. For any one couple, a history of varicocele, male genital infections or testis damage multiplies the risk of primary infertility by a factor of 28, 3 and 4 respectively, and previous female infections (salpingitis) multiply the risk by 45. Similarly, a history of ectopic pregnancy, sexually transmitted diseases or salpingitis multiplies the risk of secondary infertility by a factor of 5, 4 and 7 respectively, and a history of varicocele multiplies this risk by 4.     

南宁地区864例男性不育患者的细胞遗传学分析

目的探讨男性不育与染色体异常的关系。方法通过对2009年1月-2012年12月来我院门诊检查的864例男性不育患者进行染色体核型分析。结果864例不育男性患者中,染色体异常发生率为12．0％（104／864）,其中性染色体异常占4．5％（39／864）,常染色体异常占7．5％（65／864）。结论染色体异常是造成男性不育的一个重要因素。因此．对少精子、弱畸精子、无精子等的不育患者,进行染色体核型分析是十分必要的。     

男性不育患者Y染色体的AZF基因微缺失分析

目的探索男性不育患者Y染色体AZF基因微缺失的发生情况，为男性不育的临床诊断和治疗提供科学依据。方法应用PCR方法对36例无精或严重少精患者AZF基因的SY84、SY127和SY254进行检测分析。结果在36例患者中发现16例发生AZF基因的微缺失，均为三个基因区段不同组合的缺失。其中涉及AZFa或AZFb缺失的各8例，涉及AZFc缺失的有14例。结论AZF基因的微缺失与某些男性不育密切相关。AZFc缺失可能是男性不育中无精子、严重少精子的主要病因之一。     

男性不育的遗传学研究及在ICSI中的意义

目的在细胞遗传学基础上,建立和应用一套检测Y染色体微缺失和雄激素受体基因外显子A突变的分子诊断方法,以便研究男性不育发病机制,为临床辅助生殖技术提供遗传咨询。方法对139例原发不育患者采用外周血染色体G显带、C显带技术,并选择其中40例应用多重PCR和PCR-SSCP银染进行DAZ基因及雄激素受体基因外显子A突变检测。结果139例原发不育患者G、C显带发现54例染色体核型异常,3例DAZ基因缺失,5例雄激素受体基因外显子A发生点突变。结论通过细胞遗传学检查和DAZ基因及雄激素受体基因外显子A突变检测,从遗传学角度探讨了男性不育的病因,对卵浆单精子显微注射技术提供了理论依据。     

男性不育症患者促黄体生成素、抗精子抗体指标水平及其与精子前向运动的相关性分析

目的分析不育男性精浆促黄体生成素(LH)、抗精子抗体(AsAb)指标水平,并分析其与精子前向运动的相关性。方法选择2015年10月至2017年12月我院生殖医学科诊治的102例不育男性患者为观察组,选择同时间的42例健康育龄男性为健康对照组。观察两组对象生殖激素LH、免疫抗体AsAb指标和前向运动精子指标的差异,分析生殖激素LH、免疫抗体AsAb水平与精子前向运动的相关性。结果观察组LH水平、AsAb阳性率均高于健康对照组(P <0.05),而精子密度和前向运动精子率均低于对照组(P<0.05);不育男性患者精浆LH水平与精子密度及前向运动精子个数呈负相关(r=-0.406、-0.475;P=0.003、0.004);AsAb阳性率与精子密度及前向运动精子个数呈负相关(r=-0.518、-0.584;P=0.025、0.030)。结论不育男性患者的LH水平及AsAb阳性率较高,且与精子的前向运动密切相关,这对男性不育类型的鉴别诊断和选择治疗方案具有重要意义,应针对这种情况进行监测与干预。     

河南地区311例男性不育症患者的细胞遗传学研究

目的探讨男性不育症与细胞遗传学异常的关系。方法选取311例男性不育症患者进行外周血淋巴细胞染色体G显带核型分析。结果检出异常核型74例,占全部被检者23.79%。其中性染色体异常57例,占全部被检者18.33%,占异常核型77.03%;染色体结构异常17例,占全部被检者5.47%,占异常核型22.97%。结论性染色体数目异常是男性不育症的主要原因之一,对男性不育症患者进行染色体检查能对不育夫妇的诊断和治疗提供指导性依据。     

152例男性不育患者细胞遗传学研究

目的研究男性不育患者的遗传缺陷与精子生成障碍的关系。方法采用G显带分析152例患者外周血染色体核型,并采用计算机辅助精液分析,瑞吉染色检测精子凋亡情况,并采用聚合酶链式反应对其中染色体核型正常的患者进行Y染色体微缺失的检测。结果在152例不育患者中,检出异常核型29例,异常核型发生率为19.08%;其中60例染色体核型正常的不育患者精子凋亡率为（18.26±9.34）%,正常组为（3.52±2.11）%,两组比较有显著性差异;53例染色体核型正常的不育患者有Y染色体微缺失6例,缺失率11.3%,正常对照组未检出Y染色体微缺失。结论染色体核型异常、Y染色体微缺失以及精子凋亡是引起男性不育的重要原因。同时采用这三种遗传学检测方法可以更全面的评价男性不育患者的遗传缺陷情况,更好的为患者提供病因诊断、遗传咨询和治疗方案。     

男性不明原因不育症患者血浆中AcrAb及Sp17Ab水平变化的研究

目的探讨抗精子蛋白17抗体(Sp17Ab)、抗顶体蛋白酶抗体(AcrAb)在男性不育症发病中的作用。方法采用酶联免疫吸附(ELISA)对12例不明原因不育患者,20例其它原因引起不育的患者及25名正常的生育者的血浆中AcrAb、Sp17Ab进行检测并分析不明原因不育患者血浆中AcrAb和Sp17Ab的水平及其相关性。结果(1)血浆AcrAb水平:不明原因不育患者血浆中顶体蛋白酶抗体(4.15±1.18)μg/L,明显高于对照组的(1.49±0.51)μg/L及其它原因引起的不育(1.56±0.40)μg/L,差异有统计学意义(P0.01);对照组和其它原因引起的不育两者比较,差异无统计学意义(P0.05);(2)血浆Sp17Ab水平:不明原因不育患者血浆中精子蛋白17抗体(8.34±1.88)μg/L,明显高于对照组(4.17±1.07)μg/L及其他原因引起的不育(4.10±0.95)μg/L,差异具有统计学意义(P0.01);对照组和其它原因引起的不育两者比较,差异无统计学意义(P0.05);(3)相关性分析:血浆中顶体蛋白酶抗体与精子蛋白17抗体水平无相关性(r=0.125,P0.05)。结论血浆中顶体蛋白酶抗体、精子蛋白17抗体水平变化与不明原因不育患者发病有关。