Immunofluorescent studies on human spermatozoa. II. Characterization of spermatozoal antigens and their occurrence in spermatozoa from the male partners of infertile couples

The object of this study was to elucidate properties, organ specificity, and the occurrence in individual spermatozoa of the four antigens which can be identified in methanol-fixed human spermatozoa by the indirect immunofluorescence technique using human antisera, i.e. the antigen in the front part of the acrosome, in the equatorial segment, in the postnuclear region, and in the main tail piece.

With unfixed spermatozoa the reactions were weak and often uncharacteristic. After fixation with ethanol and acetone all four antigens were preserved, while after fixation with formalin the postnuclear antigen was not demonstrable. The antigens in the equatorial segment and in the postnuclear region were destroyed at 60°C and those in the front part of the acrosome and in the tail at 80°C. After the spermatozoa had been treated with trypsin the reaction with the three antigens in the head disappeared. However, with rabbit antiserum to human spermatozoa fluorescence could be induced even after heating the spermatozoa to 100°C or treating them with trypsin.

Absorption of the human antisera with seminal plasma, human milk, liver, kidney, and adrenal extract disclosed an antigenic relationship between the postnuclear antigen and seminal plasma as well as between the antigen in the front part of the acrosome and adrenal extract.

By means of known antisera the occurrence of acrosome antigens in the individual spermatozoa was investigated in ejaculates from seventy-six male partners of infertile couples. In ejaculates with a normal concentration of spermatozoa the antigens were generally demonstrable in at least 50% of the spermatozoa, whereas samples with a low concentration showed a marked variation, the staining percentages ranging from 0 to 90%.

     

Comparison of the occurrence of spermatozoal antibodies in male and female blood donors

Sera from 207 male and 190 female blood donors were studied for spermatozoal antibodies by sperm agglutination and indirect IFT on methanol-fixed spermatozoa.Sperm agglutinins occurred significantly more frequently in men (about 4%) than in women (0·5%), and IFT (with serum diluted 1:10) revealed a similar relation for antibodies against the main tail piece (about 10% in males vs about 3% in females). Antibodies against the front part of the acrosome and against the equatorial segment were, on the other hand, found with the same frequency in men and women, i.e. in 6–7% and 2–3%, respectively.Antibodies against the front part of the acrosome were predominantly of the IgM class, while staining of the equatorial segment and the main tail piece was mainly caused by IgG antibodies.The rare occurrence of sperm agglutinins in women may possibly be a consequence of rapid destruction of the corresponding sperm-specific antigens after ejaculation of the spermatozoa. The identical findings in men and women for antibodies against the acrosomal antigens lead to the suggestion that constitutional factors could be of particular importance for the ability to respond to these antigens.     

Immunofluorescence studies on the occurrence and localization of the CEA-related biliary glycoprotein I (BGP I) in normal human gastrointestinal tissues.

Biliary glycoprotein, I(BGP I) is a carcinoembryonic antigen (CEA) cross-reactive glycoprotein of normal human bile. Its occurrence and localization was studied in normal human gastrointestinal tissues by means of direct immunofluorescence using immunadsorbent purified BGP I antibodies with high selectivity for BGP I, as compared to CEA and 'non-specific cross-reacting antigen' (NCA). As controls fluorescein-labelled CEA and NCA were used. Specific BGP I fluorescence was only found in the biliary tract, i.e. in bile canaliculi, in the lumen of large bile ducts and on the surface of the gall bladder mucosa. No fluorescence was found in the hepatocytes or in the cells lining larger bile ducts or the gall bladder. Fluorescence probably due to cross-reaction with NCA was obtained in the cytoplasm of macrophages in different organs and on the surface of bowel epithelium.     

Use of aniline blue to assess chromatin condensation in morphologically normal spermatozoa in normal and infertile men.

Chromatin condensation is vital for the function of the spermatozoon as the motile carrier of the paternal genome. The degree of condensation can be shown with the aid of acidic aniline blue staining, which is able to discriminate between lysine-rich histones and arginine- and cysteine-rich protamines. Using this technique and employing the Düsseldorf classification of sperm morphology in cases of disturbance of spermatogenesis, it was demonstrated that chromatin condensation is impaired not only in malformed but also in morphologically normal spermatozoa. Among morphologically normal spermatozoa, the percentages of spermatozoa with chromatin condensation disturbances increase in patients with different patterns of sperm malformation, if compared with patients with normozoospermia.     

The use of enzyme-linked immunosorbent assay for detection of Mycoplasma hominis antibodies in infertile women serum samples

BACKGROUND: Besides Chlamydiae trachomatis and Mycoplasma genitalium, Mycoplasma hominis may also cause infertility due to damage of the Fallopian tubes. Therefore serum samples from infertile women were analyzed for antibodies to M. hominis. METHODS: Sera from 304 infertile women were investigated for seropositivity to M. hominis by immunoblotting and a developed ELISA. Women were classified into groups based on the type of infertility: infertile due to lack of passage in Fallopian tubes (TFI, tubal factor infertility), an infertile male partner (MFI, male factor infertility) and unexplained infertility (UFI, unexplained factor infertility). Three M. hominis isolates were used in the immunoblotting analysis and clear differences in patient immunoprofiles were observed between two isolates. For the ELISA we used a mixture of Triton X-114 extracted membrane proteins from those two M. hominis isolates as antigen. RESULTS: Ninety-seven sera (32%) were seropositive to M. hominis when tested by the ELISA. There was a significant correlation between TFI and seropositivity to M. hominis (P = 0.0015, OR = 2.21, CI = 1.35-3.61). We compared the seropositivity of 304 patients to M. hominis with the presence of antibodies against two other bacteria Chlamydiae trachomatis and Mycoplasma genitalium and there was no statistical correlation between those bacteria and M. hominis. CONCLUSION: Our results indicate that M. hominis may be an independent predictor of TFI.     

Chlamydial and gonococcal antibodies in sera of infertile women with tubal obstruction.

Sera from 48 infertile women with tubal pathology and from 77 infertile women with normal fallopian tubes were tested by enzyme linked immunosorbent assay (ELISA) using Chlamydia trachomatis and Neisseria gonorrhoeae antigens. Control sera were obtained from women undergoing abortion, sterilisation, and from women practising barrier contraception. The results of ELISA for antibodies to chlamydiae were in close agreement with results published previously of an immunofluorescence test on these sera. Antibodies to C trachomatis were found in 73% of the infertile women with tubal pathology, significantly more than in any of the control groups. Only a very low prevalence (2-5%) of antibodies to gonococcal pili was found in all groups, except women undergoing abortion (16%).     

Localization of the human SH-protease inhibitor in the epidermis. Immunofluorescent studies.

Human epidermis contains a low molecular weight SH-protease inhibitor (Human Epidermal Inhibitor = HEI), whose epidermal localization was performed with the indirect immunofluorescence method. The fluorescence was most intensive in the cytoplasms of epidermal cells, often occurring perinuclearly. The fluorescent material in the frozen sections was often finely granular and occasionally extended outside the cytoplasm, while the fluorescence in fixed sections was more uniform, but weaker. Stratum basale generally stained poorly or not at all, as did also stratum lucidum. Stratum corneum stained fairly intensively throughout. In addition to fixation, the outcome of staining was also affected by the thickness of the epidermis, particularly stratum corneum. The significance of this inhibitor for the differentiation of epidermal cells and the keratinization of epidermis has therefore been discussed, and the authors assume it to be of considerable significance in these processes.     

女性不孕与支原体、衣原体感染和抗精子抗体的相关性

目的研究支原体(ureaplasma urealyticum UU)、衣原体(chlamydia trachomatic CT)感染与抗精子抗体(antisperm antibodies ASAb)对女性不孕的影响.方法聚合酶链反应(PCR)法和ELISA法分别对128例原发性和195例继发性不孕患者的宫颈分泌物进行UU、CT和ASAb检测,同时选择健康已孕妇女96人作为对照组.结果不孕组UU、CT感染率、ASAb阳性率与对照组比均有显著性差异(P＜0.05);原发不孕组与继发不孕组UU、CT感染率、ASAb阳性率相比无显著性差异(P＞0.05);不孕组中UU、CT感染阳性患者ASAb阳性率明显高于UU、CT阴性患者(P＜0.01).结论女性生殖道UU、CT感染和ASAb与不孕有着密切的相关性,特别是继发性不孕患者尤为显著,且ASAb的产生与CT,UU感染有关.     

Infertility: Expression of heat shock protein 70 kDa in human endometrium of normal and infertile women

The expression of heat shock protein (hsp) 70 was investigatedin endometrial samples from patients with unexplained infertilityassociated (n = 5)or not associated (n = 10) with endometriosis,and compared with a control group consisting of fertile women(n= 27) with reported menstrual disturbance. The expression ofhsp, and in particular hsp 70, is up-regulated in response tomany physico-biochemical insults as well as infection and possiblyoncogenic transformation and is a good indicator of a biologicalsystem under stress. A significant over-expression of hsp 70was found in the infertile groups (P < 0.001), suggestingthat a stress response may be involved in the aetiology of unexplainedinfertility irrespective of the presence of endometriosis.     

The clinical significance of antisperm antibodies in infertile couples.

Sera from 150 women and 162 men with unexplained infertility were examined using a commercial enzyme-linked immunosorbent assay (ELISA) kit for antisperm antibodies. The results were compared to those of the Friberg agglutination test, the post-coital test, the sperm-cervical mucus contact (SCMC) test and the pregnancy rate. We also tested follicular fluids obtained from 38 women who underwent in-vitro fertilization (IVF). These data were compared with those obtained in serum, post-coital test data and with the later development of the oocyte in IVF. Antibodies in follicular fluid were found only in women with antibodies circulating in serum. The correlation coefficient between these was 0.88 (P < 0.001). There was no correlation between antisperm antibodies in serum found with the ELISA test, and with the agglutination test, the post-coital test or the SCMC test. Neither was there any correlation between antibodies in follicular fluid and the post-coital test, the pregnancy rate or successful IVF.     

两种冷冻保护剂对形态正常精子百分率影响

目的比较甘油和甘油-卵黄-柠檬酸钠(GYC)两种冷冻保护剂对精子形态影响.方法应用甘油和GYC两种冷冻保护剂(CPM)对精液标本进行速冻和缓慢冷冻保存,应用计算机精液分析仪进行精子形态分析.结果冷冻复温后形态正常精子百分率与冷冻前比较的形态正常精子百分率比较明显下降(P<0.001);速冻与缓慢冷冻法中两种保护剂间形态正常精子百分率比较均无显著性差异(P>0.05). 结论冷冻保存易造成形态正常精子百分率下降,两种保护剂对精子形态没有影响.     

The occurrence of human wart-virus antibodies in dogs,pigs and cattle

Summary Using the immunodiffusion method, antibodies against human wart-virus were detected in dog, pig and cattle sera but not in horse and reindeer sera. Antibodies were found in 25 per cent (28/114) of the dog sera, the prevalence of antibodies being fairly similar to that in children of the same age. By electron microscopy the antibodies in dog sera were shown to attach to and precipitate the human wart-virus particles, and in immunodiffusion the precipitation lines of human and dog sera were shown to be identical. Of the cattle sera studied, 10 per cent (10/97) and of the pig sera 13 per cent (5/40) gave a faint precipitation line, having identity with that given by the human sera, against human wart-virus. Conversely 18 per cent (9/50) of adult human sera contained precipitating antibodies against a bovine papilloma antigen, probably bovine papilloma virus.With 5 Figures     

不孕不育患者循环抗精子抗体阳性因素分析及治疗

作者分析了不孕不育患者中循环抗精子抗体阳性者 2 76例 ,其中男性 130例 ,与正常生育组相比 ,精液体积、密度、Mh差异无显著性 ,而精子活率、活力、白细胞阳性率、SPIM、UU差异有显著性 (P <0 .0 1)。女性患者 146例 ,与正常生育组相比 ,在黄体不健、子宫肌瘤方面差异无显著性 ,而在阴道炎症、宫颈糜烂、内生殖器炎症方面有显著性差异 (P <0 .0 1)。男女循环抗精子抗体阳性的治疗 ,单用西药组、单用中药组、中西药结合组、中西药结合合并抗感染治疗组的 3个及大于 3个月的累计治愈率分别为 9.7%、5 1.2 %、70 .6 %、89.6 %、和 19.4%、5 8.1%、82 .4%、94.8% ,各组治愈率差异均显著