输精管结扎术后抗精子抗体在非射出精子上的存在

４８例正常育龄男子和２４例要求作输精管吻合术者，用免疫珠结合试验（ＩＢＴ）和精子－宫颈粘液接触试验（ＳＣＭＣ）分别检测其近睾端输精管液中的精子（非射出的精子）表面抗精子抗体的情况。结果显示：正常育龄男子非射出的精子上没有抗体存在；而输精管结扎术后非射出的精子上有抗精子抗体存在（ＩｇＧ：７９．４％，ＩｇＡ：３８．２％，ＳＣＭＣ：３５．５％）。这些抗体在输精管结扎术后不到１年就可发现，并保持到结扎术后３年以上。吻合术后１～３个月内，其精液中射出的精子上仍发现抗体存在。结果还提示：输精管结扎术后，不能以非射出的精子上抗体的水平去预测吻合术后射出的精子上抗体的水平；吻合术前血清抗体水平与非射出的精子上抗体水平并不一致。     

Vasectomy and vasovasostomy in rhesus monkeys: the effect of circulating antisperm antibodies on fertility.

Rhesus monkeys develop agglutinating and complement-dependent antisperm antibodies after vasectomy. In order to study whether these antibodies affect fertility after vasovasostomy, 15 animals were given vasectomies and 6 months later vasovasostomies. Subsequently, each was mated with females of proven fertility. Five controls were given sham operations and similarly treated. During this period, each aimal was bled for serum to monitor the humoral immune response, ejaculated for semen analyses, and palpated for granuloma or fistula development. All control animals had a transient decrease in sperm density after sham vasectomy and vasovasostomy operations. The surgical procedures of vasectomy and subsequent vasovasostomy resulted in more animals having sperm of poor motility and quality. All of vasovasostomies were surgically successful in that sperm were again present in the ejaculate of each animal. The amount of sperm in the ejaculate could not be correlated with the ease of surgical procedure, presence or absence of macrophages in the ejaculum, motility, or forward progression. Only animals that had been vasectomized developed circulating antisperm antibodies. Sustained, elevated levels of antisperm antibodies most commonly occurred in monkeys that had high initial total sperm counts. Six of the experimental animals retained high levels of sperm-immobilizing antibodies after vasovasostomy. Of these, two were found to be infertile and two were classed as subfertile. Of the nine experimental animals without sustained antisperm antibody production, only one was classed as subfertile. This suggests that antisperm antibodies may in some cases impair the restoration of fertility after vasovasostomy.     

Low fertility rate in vasovasostomized males and its possible immunologic mechanism.

Contradictory views have been expressed about the role of the various antisperm antibodies which develop after vasoligation. The present study was conducted in 50 normal fertile males, 50 vasectomized subjects and 25 subjects after recanalization of their vas deferens in order to investigate the development of various anti-sperm antibodies after vasectomy, along with their incidence, their persistence after successful relief of vaso-obstruction by vasovasostomy and their role in the causation of infertility in vasoanatomized normospermia males. Sperm agglutinating, immobilizing and haemagglutinating antibodies showed rises in titres with increase during the post-vasectomy period, indicating continuous antigenic stimulus. Age, post-operative complications and blood group did not seem to alter the results. 86% of subjects developed antisperm agglutinins, mostly tail-to-tail type (54.5%), 1-12 years after vasoligation, while only 2% of fertile men had circulating spermagglutinins. A lower incidence of positive sperm in the immobilization test than in the agglutination test suggests either that different antibodies are detected by these two tests or these tests have differing sensitivities. Of the 25 vasovasostomized subject, 13 (52%) cases became normospermic and 4 (16%) oligospermic while 8 (32%) remained azoospermics. Except for 3 oligospermic subjects, all had circulating spermagglutinins. Among the 13 normospermic vasovasostomized persons, a significant correlation was found between the titres of circulating antisperm agglutinins and autoagglutination of spermatozoa in their ejaculates; and also between the sperm immobilization values of their sera and the degree of their sperm motility. Three normospermic recanalized men, having low levels of sperm agglutinins and haemagglutinins with normal seminogram and no sperm immobilizing antibody, successfully impregnated their wives. Another 10 vasovasotomized infertile subjects had sperm agglutinins in significant titre; 5 showed positive sperm immobilization values, a similar number showed autoagglutination of sperm, while a decreased degree of motility of sperms was noted in 6 cases. Thus there was a significant correlation between the titres of anti-sperm antibodies and autoagglutination of spermatozoa, which might be an important cause of male infertility after successful anatomic relief of vasoobstruction. Histological studies of testicular biopsy showed normal spermatogenesis in azoospermic recanalized subjects, although they had high levels of antisperm antibodies. This suggests that these antibodies do not affect normal spermatogenesis, and sperm counts.     

Factors that influence fertility after vasovasostomy in rats

OBJECTIVE: To determine if fertility after vasovasostomy of immunologically responsive Lewis rats differs from that of the less responsive Sprague-Dawley strain and to relate fertility to antisperm antibodies, fluid flow in the vas deferens, and testicular structure. DESIGN: Male rats received: (1) bilateral vasectomies; (2) vasectomies followed 3 months later by vasovasostomy; or (3) sham operations. SETTING: Research laboratory. MAIN OUTCOME MEASURES: Fertility was assessed by caging males with three females for 2 weeks and subsequently counting implantation sites. Antisperm antibodies were measured with an enzyme-linked immunosorbent assay, fluid flow through vas deferens segments was tested in vitro, and testicular structure was studied microscopically. RESULTS: Nearly all vasovasostomized Lewis rats were infertile (33 of 34), whereas 62% (18 of 29) Sprague-Dawley rats were fertile after vasovasostomy (P less than 0.001). In fertile Sprague-Dawley males, significant correlations existed between: (1) implantation sites or females impregnated; and (2) antisperm antibodies early after vasectomy, vas flow, and testicular morphology. CONCLUSIONS: Genetic differences affect fertility after vasovasostomy. Fertility after vasovasostomy is also influenced in a multifactorial manner by the immune response, mechanical elements, and structural changes in the reproductive tract.     

Human sperm protein encyclopedia and alloantigen index: mining novel allo-antigens using sera from ASA-positive infertile patients and vasectomized men

Anti-sperm antibodies (ASA) are an important cause of immunological infertility. The objective of this study was to identify immunodominant sperm antigens recognized by anti-sperm antibodies (ASA) in serum samples of infertile men, women and vasectomized men. High-resolution two-dimensional gel electrophoresis was employed to separate human sperm proteins using isoelectric focusing (IEF) or nonequilibrium pH gradient electrophoresis (NEPHGE), followed by PAGE and Western blotting. Serum samples from five infertile male and five infertile female subjects that contained ASA as assayed by the immunobead binding test (IBT), were analyzed by Western blotting using NEPHGE gels followed by enhanced chemiluminescence (ECL) to identify the basic sperm antigens reactive to the sera. Serum samples from five fertile male and five fertile female subjects that were ASA-negative by IBT were used as controls. Serum samples from six vasectomized men collected before vasectomy and at different time intervals until 6 months after vasectomy were analyzed by Western blotting using IEF gels. The ECL blots were analyzed to compare immunoreactivity between serum samples from fertile and infertile subjects and identify antigens unique to sera of the infertile subjects. Similarly, immunoreactivity between serum samples from pre- and post-vasectomy was compared to identify antigens unique to sera collected following vasectomy. Five allo-antigenic basic protein spots were recognized by sera from infertile males but not from fertile subjects. Five sperm iso-antigenic basic spots were recognized by infertile female subjects. Two among six of the vasectomized men's sera showed a difference in the Western blot profile 6 months after vasectomy, recognizing at least one new protein spot in each case when compared to pre-vasectomy sera. The acrosomal protein SP-10 was identified as an alloantigen recognized by a post-vasectomy serum. Molecular identities of the known allo- and iso-antigens identified in this study and in previous studies from this laboratory are reviewed and discussed.     

输精管吻合家兔不育原因的研究——Ⅰ.血清抗精子抗体分析

本实验证明,输精管吻合术后不育组(VIG)血清抗精子抗体阳性检出率和效价与输精管吻合育组(VFG)比较无差异(p>0.05);VIG的精子密度显著地低于VFG(p<0.01)和假手术组(SOG)p<0.001;精子膜ADA活力,VIG明显地低于SOG(p<0.01),VFG与SOG比较无差异(p>0,05);精子膜ADA活力与精子密度呈显著的负相关(r=-0.60,p<0.01);睾丸ACE活力与精子密度呈正相关(r=0.59,p<0.01),而与ADA活力呈有意义的负相关(r=-0.52,p<0.02)。因此,吻合术后不育与血清抗精子抗体效价无直接关系,而与精子密度降低、功能障碍有关。     

Vasectomy: consequences of autoimmunity to sperm antigens.

Data from studies examining the effects of vasectomy in a large number of nonhuman primates vasectomized for periods ranging up to 14 years are summarized, and these findings and speculations are used as a framework with which to review the subject of autoimmunity and vasectomy. Attention is directed to autoimmunity to sperm antigens following vasectomy (factors affecting antisperm antibody levels, characteristics of circulating antisperm antibodies, antisperm antibodies in seminal plasma, and cellular immunity following vasectomy), and immunopathology of antisperm autoimmunity (local effects on the male reproductive tract and systemic effects on the male reproductive tract). The 6 hypotheses that have been advanced to explain individual variations in dynamics and types of antisperm antibodies produced following vasectomy are reviewed. 3 tests are commonly used to detect free antisperm antibodies after vasectomy: 1) the spermagglutination test; 2) the sperm immobilization test; and 3) the immunofluorescence test. Spermagglutinating (SA) antibodies, the most common type of antibody produced after vasectomy, occur in approximately 2/3 of vasectomized men and in a majority of vasectomized rhesus monkeys. Sperm-immobilizing (SI) antibodies are also produced in a large percentage (40%) of vasectomized men and rhesus monkeys. About 30% of vasectomized men also have antiprotamine antibodies.     

Human sperm function in the ejaculate following vasectomy

Semen specimens were obtained for analysis from nine men immediately before vasectomy and 3 days following the vasectomy procedure. After vasectomy there was a significant decrease in seminal sperm concentration, percentage of sperm motility, and mean swimming speed. In seven of nine cases the spermatozoa did not survive overnight incubation in capacitation media, and in no case was penetration of zonafree hamster oocytes observed 3 days after vasectomy.     

Open-ended vasectomy: an assessment

Leaving open the testicular end of vas at vasectomy could reduce symptoms of epididymal congestion and improve the success rate of vasovasostomy but might have the disadvantage of increasing the incidence of painful sperm granulomas and spontaneous recanalization. In 4330 open-ended vasectomies the rate of epididymal congestion was significantly less than in 3867 standard vasectomies. The rate of painful sperm granulomas was not increased: it was significantly reduced. Spontaneous recanalization was rare in both groups. Whether or not open-ended vasectomy improves the success rate of vasovasostomy, it represents an improvement in technique because it reduces the rate of complications after vasectomy. Closure of the sheath over the prostatic end of vas is essential if recanalization is to be prevented.     

Clinical experience of epididymovasostomy and vasovasostomy for obstructive azoospermia]

14 patients with obstructive azoospermia were treated with microsurgical vasovasostomy or epididymovasostomy. A total of 11 patients underwent 2-layer vasovasostomy, 6 cases following bilateral vasectomy and 5 cases suffering from bilateral vasal disruption by inguinal herniorrhaphy. 3 patients underwent end-to-end epididymovasostomy with Silber's specific tubule technique following bilateral epididymitis. After the operation, fertility was restored in 6 of 14 cases. The recovered sperm density was greater than 20 x 10 6/ml and sperm motility exceeded 50%. Pregnancy was achieved in 1 case following inguinal herniorrhaphy. Preoperative testicular biopsies carried out in 11 cases disclosed slight deterioration of spermatogenesis. Johnsen's mean score was 8.20 +or- 0.38 (mean +or- S.D.). The mean scores of the biopsies obtained from the cases obstructed for more than 20 years were significantly lower than those with less than a 20-year time period. After inguinal herniorrhaphy, reanastomosis of the vas deferens was difficult in order to restore fertility. This was due to the presence of longterm obstruction, defects in the vas deferens, or the absence of sperm granuloma at the vasectomy site. Epididymovasostomy restored fertility in 67% of the cases. The specific tubule technique was the most effective in the management of such pathological conditions. (author's modified)     

Characterization of humoral antibodies reactive with spermatozoa, N-acetyl galactosamine, and a putative blood group antigen in seminal plasma

Antisperm antibodies in sera of infertile women may react differently with spermatozoa of different men. We studied the reactivity of these antibodies with spermatozoa from men of varying blood group status. Increased immunoglobulin binding to sperm of group A or AB men was noted when compared with group O men. A diminution in binding of immunoglobulins to spermatozoa after absorption of these sera with human group A or AB red blood cells was noted as well as after coincubation of sera and sperm with N-acetyl galactosamine, the terminal sugar of blood group antigen A. These observations suggest that antibodies directed against blood group antigens adsorbed to sperm of secretor males may account in part for variations in immunobead binding levels between sperm of different men.     

Lack of association of the development of anti-sperm antibodies and other autoantibodies as a consequence of vasectomy.

The possibility of autoantibodies--other than sperm antibodies--developing as a consequence of vasectomy has been investigated in 255 volunteers. During the first year after vasectomy no obvious increase was observed in the occurrence of any of these antibodies (rheumatoid factor, antinuclear antibodies and antibodies against smooth muscle, mitochondria, gastric parietal cell, thyroid microsomes and thyroglobulin). Ninety-nine of the patients were also examined for agglutinating and immunofluorescent antibodies to sperm to see if there was any relationship between the occurrence of anti-sperm antibodies and other autoantibodies. However, the prevalence of non-sperm autoantibodies did not differ in two nearly equal groups of patients with and without indications of autoimmune reactions to spermatozoa, respectively. Consequently the present results lend no support to the hypothesis that vasectomy could induce autoimmunity to other autoantigens than sperm-specific antigens.     

Sperm antibody production in female sterility

A review of the immunological implications in reproductive physiology is presented. Although attempts have been made to ascribe the antigenicity of semen to individual components, it has not been possible to isolate the human semen antigen responsible for infertility. In monkeys total ejaculates and seminal plasma have shown higher antigenicity than washed spermatozoa. In bulls some evidence of such antigens have been found in the seminal plasma. They are iron-binding proteins resembling lactoferrin. Most investigators have found no evidence for any participation of the ABO blood group antigens in cases of sterility. On the surface of human spermatozoa histo-incompatibility antigens have been detected. Transplantation antigens may be related to sterility. However, an immulogic tolerance of the maternal organism exists against the genetically foreign fetal tissue. Autoimmune spermagglutinating antibodies have been detected in the sera and in the seminal plasma of males with sterility. An obstruction of the seminal pathways may facilitate the production of such antibodies against retained sperm. Isoimmunity in females against seminal components has been shown in cases of sterility; however, fertile women have also been shown to have such conditions. In a group of infertile women spermagglutination activity was detected in 7.5% of cases. In another series of 46 cases with primary unexplained infertility agglutinating antibodies were found in 17.4%. Other investigators have also reported higher rates than the authors. The sperm immobilization test seems to be more sensitive than the agglutination test. No sera were found positive with both tests. With immunofluorescent techniques humoral sperm antibodies have been found to be the IgM and IgG fractions. Each acts on a different part of the spermatozoa. The only promising therapy against humoral sperm antibodies is avoidance of sperm contact over a long period of time. Reported results have been conflicting. Cortisone did not improve the result. Desensitization or immunosuppresive efforts are of uncertain value.     

Epididymal extravasation following vasectomy as a cause for failure of vasectomy reversal.

Twenty-eight men undergoing vasectomy reversal who were found to have no sperm in the proximal vas fluid on one or both sides underwent microscopic epididymal exploration. In 33 of 39 cases so explored, normal sperm were found in the epididymal fluid of the corpus, despite absence of sperm in the vas fluid. Epididymal histology distal to this site revealed extensive interstitial sperm granulomas resulting from rupture of the epididymal duct. Testicular biopsy revealed normal spermatogenesis. Secondary epididymal obstructions were noted when there was copious fluid in the vas deferens proximal to the vasectomy site as well as when there was scanty fluid. It is concluded that persistent azoospermia after an accurate microscopic vasovasostomy results from the secondary epididymal obstruction induced by rupture of the epididymal duct related to the pressure increase after vasectomy.     

Induction of infertility in adult male bonnet monkeys by immunization with phage-expressed peptides of the extracellular domain of FSH receptor

Active immunization of proven fertile adult male bonnet monkeys (Macaca radiata) with phage-expressed follicle-stimulating hormone receptor (FSHR)-specific peptides from the extracellular domain resulted in a progressive drop in sperm count with all animals becoming azoospermic by day 100. However, serum testosterone concentrations were unaltered during the entire course of study and animals exhibited normal mating behaviour. Breeding studies with proven fertile female monkeys revealed that all the immunized males were infertile. Following interruption of immunization on day 225, sperm counts returned to normal with restoration of fertility. These results indicate that infertility can be induced in adult male monkeys by interfering with the action of FSH using specific peptides of the extracellular domain of FSHR as antigens, without the risk of producing cross-reacting antibodies to the other glycoprotein hormones.     

Effect of vasovasostomy on contralateral testicular damage associated with unilateral vasectomy in mature and immature Lewis rats

OBJECTIVE: We sought to determine if laser-assisted vasovasostomy could reverse the contralateral histologic testicular changes associated with unilateral vasectomy. DESIGN: A prospective, randomized, blinded, controlled study. SETTING: Animal microsurgical laboratory, St. John's Mercy Medical Center, St. Louis, Missouri. PATIENT(S): Twenty mature and 20 immature male Lewis rats. INTERVENTION(S): Ten mature and 10 immature male Lewis rats underwent unilateral vasectomy. At 5 months, testicular biopsy and laser-assisted vasovasostomies were performed followed 2 months later by evaluation of vas patency and repeat testicular biopsy. Control animals consisted of 10 rats in each group, 5 that underwent sham operations and 5 that had halothane anesthesia alone. RESULT(S): In the immature and mature groups unilateral vasectomy resulted in marked contralateral testicular damage in 30% (3 of 10) and 50% (5 of 10), respectively. Vas patency determined 2 months after vasovasostomy was 80% (8 of 10) in the mature group and 89% (8 of 9) in the immature group. No animal that had contralateral testicular changes after vasectomy and a patent vas after vasovasostomy showed improvement in testicular histology. CONCLUSION(S): It appears that contralateral testicular damage associated with unilateral vasectomy is not improved 2 months after successful vasovasostomy in mature or immature Lewis rats.     

Some gross observations of the epididymides following vasectomy: a clinical study.

In 114 vasectomized men, 220 epididymides were grossly examined during vasovasostomy. Fullness, distention, turgidity, thickening, induration, and other gross changes of the epididymides, including the formation of cystic spermatic granuloma, or spermatocele, indicated inadequate removal of spermatozoa and testicular fluid from the sequestrated proximal seminal ducts and the epididymis. Further studies are necessary to determine whether the epididymal changes following vasectomy have any effect on sperm morphology and sperm physiology. The information obtained from these studies possibly may explain the poor pregnancy rates in the spouses of successfully vasanastomosed patients.     

Infertility in mice after unilateral vasectomy

The effects of unilateral vasectomy upon fertility and antisperm antibody production were studied using DBA/1J mice. Thirty-six males underwent either unilateral vasectomy, unilateral orchiectomy, or sham surgery. In vivo effects upon fertility were monitored by matings. Antisperm antibody titers were measured monthly. In vitro fertilization was performed in the presence of serum obtained 4 months postoperatively, and serum testosterone levels were also determined. After 3 months, only 1 male in the vasectomy group induced a pregnancy (1 of 12), while all but 1 of the males in the two control groups induced a pregnancy (20 of 21). The geometric mean antisperm antibody titer was 1:169 in the vasectomy group, while the orchiectomy and sham surgery groups had titers of 1:4 and 1:14, respectively (P less than 0.0001). The percentage of eggs fertilized in vitro in the presence of serum from experimental mice fell from 80% in the two control groups to 40% in the unilateral vasectomy group. Unilateral vasectomy induced infertility in DBA/1J mice and an antisperm antibody response. Sera containing these antibodies inhibited in vitro fertilization. This suggests that infertility after unilateral vasectomy may be immunologically mediated by antisperm antibodies.     

Vasectomy failure using an open-ended technique

An open-ended vasectomy technique that involved purposeful creation of a sperm granuloma on the testicular side of the vas was used in 4 patients as a possible contribution to easier reversal, since animal and human studies suggest that such a granuloma prevents pressure-induced epididymal damage and favors maintenance of normal spermatogenesis. The 4 vasectomies were performed at an outpatient clinic using local anesthesia. Through transverse high scrotal incisions, .5 cm segments of vas were removed, and the abdominal side lumina were cauterized to a depth of .5 cm. The testicular side was neither cauterized nor ligated. No complications occurred in any patient, and all wounds healed normally. Examination of semen specimens 15 ejaculations after vasectomy revealed azoospermia in 2 men and active motile sperm in 2 others. Repeat semen analyses on the 2 men up to 5 months after vasectomy revealed persistence of motile spermatozoa. Repeat vasectomies were preformed, at which time sperm granulomas .5 cm in diameter were excised, and both vas lumina were cauterized, resulting in azoospermia on later semen analysis. The failure rate of 50% with the open-ended technique is clearly unacceptable. Cauterizing or removing a longer length of vas and transposition of the open testicular end to a separate fascial plane might reduce failure rates to acceptable levels, but until this is proven in large-scale studies, vasectomy should be presented as a permanent operation.     

输精管结扎与吻合后人精浆L－肉毒碱果糖含量及α－1、4糖苷酶活力研究

本文报道了６３例接受显微外科输精管吻合术患者术前和４８例术后１～１２个月精浆α－１、４糖苷酶活力，Ｌ－肉毒碱和果糖含量的结果，并以４７例正常生育男子作对照研究。输精管结扎后男子精浆α－１、４糖苷酶活力与Ｌ－肉毒碱合量明显低于对照组。在吻合术后随访期间，约２／３男子输精管复通良好，精子密度（　），精浆α－１、４糖苷酶活力（４２．３６±３１ｍＩＵ／ｍｌ）和Ｌ－肉毒碱含量（４７１．８５±１９４．７１ｎｍｏｌ／ｍｌ）均达正常水平。１／３男子精浆α－１、４糖苷酶活力、Ｌ－肉毒碱含量无明显增高，同时表现出少精子（５．３±４．２×１０６／ｍｌ）与无精子。