The immunobead test as a sperm antibody detector

The occurrence of sperm antibodies on the surface of sperm can be demonstrated by several methods. A study to evaluate the immunobead (IB) test as a laboratory procedure during the detection of sperm antibodies among infertile males showed an overall prevalence of 6.8% and 5.8% for IgG and IgA respectively--convincing evidence that the IB test can be employed as a sperm antibody detector.     

Antisperm antibodies detection with the immunobead test as viewed by TEM

The immunobead test (IBT) was used for the detection of antisperm antibodies. Scanning electron microscopy (SEM) was used to study ultrastructural aspects of this technique. This brief communication aims to present some ultrastructural evidence using transmission electron microscopy (TEM).     

The detection of antisperm antibodies in serum: a comparison of the tray agglutination test, indirect immunobead test and indirect Spermcheck assay

Testing for antisperm antibodies (ASAs) is an important part of the work-up of the sub-fertile couple, yet there is little consensus regarding the most appropriate methods. The Spermcheck assay (GSC; Bio-Rad Laboratories Inc., Diagnostics Division, Hercules, CA, U.S.A.) is supplied with wash buffer, controls and bead reagent which detects all three major classes of ASAs (IgA, IgG and IgM) in a single test. This study compared results on a bank of samples using the tray agglutination test (TAT), immunobead test (IBT), GSC and a modified Spermcheck assay to detect a single isotype in each test (SISC). The IBT and SISC showed excellent correlation, with 127/141 (90.1%) tests agreeing. There was an apparent lack of sensitivity to IgM with GSC as 8/15 (53.3%) samples testing positive with IBT and 7/15 (46.7%) testing positive with SISC were negative with GSC. Of the 24 IBT-negatives, seven (29.2%) were positive for TAT, indicating a high incidence of non-immunological agglutination, though this decreased as the TAT titre increased. The proportion of samples testing positive for IBT increased with TAT titre: 3/20 (15.0%) for TAT-negative samples, 6/10 (60.0%) for low titres and 21/24 (87.5%) for high titres. This was also observed when comparing the GSC with TAT. The TAT therefore appears useful as a first-line screen, whilst the inability of the GSC to adequately detect IgM limits its use as an indirect test. Both the IBT and SISC can be used to further investigate the type and class of ASA present.     

Testicular alterations are linked to the presence of elevated antisperm antibodies in Sprague-Dawley rats after vasectomy and vasovasostomy

The relationship between alterations in testicular histology and antisperm antibodies was studied after vasectomy and vasovasostomy in Sprague-Dawley rats, which are immunologically relatively non-responsive to vasectomy. Testes were prepared for histologic study at intervals up to seven months after vasectomy, vasectomy followed three months later by vasovasostomy, or sham operations. Antisperm antibodies were assessed with an ELISA. Testicular alterations, which were observed in a minority of animals after vasovasostomy, consisted mainly of depletion of germ cells. Mean serum antisperm antibody levels were greater for animals with altered testes than for rats with normal testicular histology. In addition, the proportion of rats that showed a positive antisperm antibody response was greater among animals with testicular changes than among those with unaltered testes. When the present results on Sprague-Dawley rats were compared with previous findings on the highly responsive Lewis strain, it was evident that the incidence of testicular changes and the proportion of positive antibody responders were greater in the Lewis strain. However, elevated antisperm antibodies and testicular alterations appeared to be more tightly linked in the less responsive Sprague-Dawley rats.     

Immunological assessment of renal cancer patients as evaluated by the lymphocyte adherence inhibition test

The lymphocyte adherence inhibition test was used to evaluate tumor immunity toward 2 types of soluble renal cancer antigens extracted from 3 different renal cancer specimens. These extractions were accomplished with either 3 molar potassium chloride or 2.5 per cent butanol, and were tested in 23 patients with renal cancer, 8 with benign renal disease and 9 with bladder cancer. In 62 tests of lymphocyte adherence inhibition reactivities toward 3 molar potassium chloride in renal cancer patients the mean value of lymphocyte adherence inhibition reactivity (32.7 +/- 15.4 per cent) was significantly greater compared to that observed in 20 examinations in patients with benign renal disease (17.6 +/- 9.4 per cent, p less than 0.001) and 24 tests in bladder cancer patients (15.4 +/- 4.4 per cent, p less than 0.001). With a lymphocyte adherence inhibition index of greater than 20 per cent as a positive response, there were 50 true positive findings (81 per cent) in the renal cancer group, while the combined benign renal disease and bladder tumor groups had 34 true negative responses (77 per cent). With the butanol extract the mean lymphocyte adherence inhibition reactivity of the 62 tests done on renal cancer patients (38.9 +/- 10.8 per cent) was significantly greater than either control group (7.1 +/- 6.2 per cent, p less than 0.001 for benign renal disease and 5.9 +/- 2.6 per cent, p less than 0.001 for bladder tumor). Moreover, of 62 tests done on renal cancer patients 59 (95 per cent) had true positive responses, while 43 of the 44 tests (98 per cent) in the combined control group showed true negative findings. These results suggest that immune responses in renal cancer can be assessed with the lymphocyte adherence inhibition test, and they may be useful in the diagnosis and management of renal cancer patients.     

Immunological state of patients with carcinoma of the bronchus before and after radiotherapy.

The immunological state of 30 patients with carcinoma of the bronchus was assessed before and after radiotherapy by lymphocyte response to PHA and E and EAC rosette formation. The results were compared with those from age-matched patients with benign chest disease and a group of healthy control subjects. Differences were found between the three groups and decreased immunological responses were found to correlate with shorter survival times for patients with cancer of the bronchus. These differences were not associated with the extent of the disease, or with the smoking habits of the patients. Significant differences in percentage EAC cell rosetting were demonstrated between lymphocytes from patients with malignant disease (31.3 +/- 2.0) and those for control groups (21.5 +/- 1.9 and 24.0 +/- 2.2). Cancer patients and benign chest disease patients both had significantly decreased mean E rosetting values (59.3% and 55.6%) compared with healthy control subjects (69.7%). The group of cancer patients with a normal percentage of T lymphocytes and total number of lymphocytes after radiotherapy, or those with low percentage EAC cell rosettes, had a greater than 80% survival after seven months compared with less than 50% for the rest of the patients with carcinoma of the bronchus.     

男女性不育症8331例血清抗精子抗体检测结果分析

目的 观察男性和女性血清抗精子抗体的阳性率、滴度及其差异性.方法 样本获自男科和妇科就诊的不育患者,血清分离后用精子浅盘凝集试验(TAT)和精子制动试验(SIT)检测,并用已知阳性和阴性血清作质控对照.结果 男女性不育症8331例中,抗精子抗体阳性率26.8%,其中男性25.2%,女性29.9%,其差异有统计学意义(x2=20.13,P<0.005).抗精子抗体阳性的2232例中,单一TAT阳性90.4%,TAT和SIT均阳性9.6%,无单独SIT阳性检出.TAT阳性率,男性不育症为25.2%,女性不育症为29.9%,其差异有统计学意义(x2=26.29,P<0.001).SIT阳性率,男性不育症为3.3%,女性不育症为1.3%,其差异有统计学意义(P<0.001).结论 免疫性不育在不育症诊疗中占有非常重要的地位.     

Immunological status in three patients, thirty years after living related renal transplantation: antibody production in long-term survivors

There have been few studies of the immune status of long-term follow-up patients after living-donor kidney transplantation. We investigated the immune status from the immunologic and pathologic standpoint of three long surviving recipients who had received renal grafts more than 30 years earlier. Anti-HLA antibodies that had not been present before transplantation were detected in one recipient with three HLA mismatches. One recipient with identical HLA showed positive for the crossmatch test, but not for the panel reactive antibody test (PRA), thus showing that this patient had HLA antibodies against HLA minor histocompatibility antigens, etc. Only one patient with established microchimerism was stable without any antibody production. Pathologically, chronic allograft nephropathy with C4d staining suggestive of antibody-mediated rejection was observed in both patients with HLA antibodies. Physicians should clinically manage patients by always bearing in mind the presence of anti-donor antibodies during long-term regular follow-up of transplanted kidneys.     

γ-氨基丁酸对抗精子抗体阳性患者精子顶体反应的影响及机制探讨

目的 研究γ-氨基丁酸(gamma-aminobutyric acid,GABA)对正常人类精子及抗精子抗体(AsAb)阳性精子顶体反应的影响及其机制。方法 两组各18例采用三色染色法检测精子顶体反应率。结果 GABA增加正常及AsAb阳性患者精子顶体反应率,与对照组比较,差异显著(P<0.01);GABA可使精子的Na~ —K~ —ATPase,Ca~(2 )—ATPase的活性增加(P<0.01;P<0.05);并且显著减少精子中的MDA含量(P<0.01),抑制氧自由基的产生。结论 GABA可明显提高正常人及AsAb阳性患者精子顶体反应。     

抗精子抗体对妇女不育的影响

对１９８６年１２月～１９９２年１２月期间门诊的３９１对不育夫妇检测ＡｓＡｂ的结果进行分析，发现不育≤２年的夫妇以男方ＡｓＡｂ阳性率（４３％）最高。经过治疗后不育年限越短妊娠率越高（不育≤２年的夫妇妊娠率为３６．６％）。在１９８对ＡｓＡｂ阳性的夫妇中，治疗后转阴率为３４．８％，妊娠率为２４．２％，与双方ＡｓＡｂ阴性的不育夫妇妊娠率（２３．２％）相近。在女方不育的原因中，以生殖器官炎症者ＡｓＡｂ阳性率最高（７１．２％），其次为习惯性流产（６０％）。治疗以中药抑抗汤为主，具有温阳、养气补血、活血化瘀，以及时对免疫功能有双向调节作用。     

Dynamics of the lysosomal-cation test in patients with appendicitis before and after surgery

The diagnostic informative value of the lysosomal-cation test (LCT) in patients with appendicitis was studied. The formula of lysosomal-cation index has been suggested, peculiar features of cytomorphology in given pathology described. The LCT was established to be an informative index of form and severity of the vermiform process inflammation.     

输精管结扎术后痛性结节和附睾郁积症与抗精子抗体的关系

本文采用Kibrick、Franklin-Dukes和Isojima三种试验法,对输精管结扎术后无并发症者253例,有痛性结节和附睾郁积症者20例,以及未作输精管结扎的正常对照者319例的血清进行了检测。其结果显示输精管结扎术后这三种方法的阳性率和滴度都有非常显著的提高,但在输精管结扎术后有和无痛性结节及附睾郁积症两组间,这三种方法的阳性率无显著差异。在有这两种并发症组中,Franklin-Dukes试验的滴度高而kibrick和Isojima试验的滴度却低。因此,测定血清中的抗精子抗体对诊断痛性结节和附睾郁积症以及估价其严重程度是无助的。     

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)