Females' isoimmunity to sperm is associated with sperm autoimmunity in their husbands

One hundred twenty-five fertile couples and 334 infertile couples were tested for the presence of cytotoxic and hemagglutinating antibodies to sperm. Elevated titers of sperm antibodies were absent in both partners of fertile couples. Of 79 infertile males with levels of sperm antibodies in the previously established negative range, 97% had wives who also had low titers of sperm antibodies. Of 255 infertile males positive for serum hemagglutinating antibodies, 56% had wives whose serum contained significant circulating hemagglutinating antibodies, while 93 of 202 (46%) males with significant cytotoxic antibody titers had wives whose serum contained elevated cytotoxic antibody titers. The females developed elevated titers of sperm antibodies in the serum and cervical mucus if their husbands had significant titers of hemagglutinating and cytotoxic sperm antibodies in the serum and seminal plasma samples. Females' isoimmunity to sperm was significantly associated with their husbands' autoimmunity to sperm and infertility.     

Sperm Immunity in Infertile Couples: Antibody Titers Are Higher Against the Husbands' Sperm Than to Sperm From Controls*

ABSTRACT: Titers of hemagglutination and cytotoxic antibodies to sperm were determined in the sera, seminal plasma, cervical mucus, and vaginal secretions from 69 infertile couples, using sperm from the husbands and from normal control subjects. Titers of hemagglutination and cytotoxic antibodies were significantly higher (p = 0.002 and p < 0.001, respectively) against autologous sperm as contrasted with sperm from control subjects in sera from 55 autoimmune males. Hemagglutination and cytotoxic antibody titers were also significantly higher (p = 0.044 and p < 0.001, respectively) against their husbands' sperm as contrasted with control sperm in sera from 46 females with isoimmunity to sperm. Ninety percent of males and females with sperm immunity were positive for histocompatibility antigens HLA-B7, HLA-B8, and/or HLA-BW35. In males, the presence of increased serum cytotoxic antibody titers against autologous versus control sperm was significantly associated with the presence of HLA-B7 allele (p = 0.0017) and with HLA-B7, HLA-B8, and/or HLA-BW35 in general (p < 0.05). In the females, increased serum antibody titers to their own husbands' versus control sperm were not preferentially associated with HLA-B7, HLA-B8, or HLA-BW35 antigens.     

Study of sperm antibodies in the sera of vasectomized males

Agglutination and immobilization tests were performed to detect the presence of sperm antibodies in the sera of 100 vasectomized men. Overall, 52% of subjects had sperm antibodies. 50% showed agglutinating antibodies, while immobilizing antibodies were present in 40%. Of the 52 positive cases, 23 showed agglutinating antibody alone in their sera. 73 had both agglutinating and immobilizing antibodies, and 3.8% had sperm immobilizing activity alone. The age incidence of the presence of sperm antibodies was as follows: under 35 years, 66%; 36-40 years, 50%; 41-45 years, 50%; and 46 years and above, 33.3%. The highest percentage of presence of sperm antibodies was recorded in men who had been vasectomized less than 1 year previously (75%). Significantly high titers of sperm agglutinating activity were found in 20% of the positive cases, while high titers of sperm immobilizing activity were noted in 40% of positive cases. It has been suggested that the development of sperm antibodies after vasectomy may prevent subsequent successful restoration of fertility by reanastomosis.     

输精管结扎术后附睾精子及其吻合术后再现精子超微结构的观察

目的观察输精管结扎术后附睾精子和吻合术后再现精子超微结构的演变情况及其相关关系。方法15例要求作输精管吻合术的对象,手术时取出附睾精子（A组）并追踪其吻合术后精液再现精子（B组）,同时应用光镜和电子显微镜分别观察A组、B组的精子形态和结构的改变,并进行统计学分析。结果光镜分析显示：B组与A组比较,精子密度和尾畸比例下降有显著性;而精子活动率和头畸比例升高有显著性;正常形态的比例有增高的趋势。电镜分析显示：B组与A组比较,正常形态精子显著升高,而尾部畸形精子明显降低,两者差异均有统计学的显著性;头畸形和颈畸形的比例在两组间差异没有显著性;而两组间尾部畸形比例呈正相关。透射电镜显示内部结构在头、颈、尾均有多细胞器、多种形式的改变。结论本文用电镜观察进一步证实了输精管结扎术后附睾精子形态向输精管吻合术后精液再现精子变化的规律。不论电镜还是光镜分析,精子尾部异常都是由高比例（A组）向低比例（B组）转化,正常形态的精子均由低比例（A组）向高比例（B组）转化;精子头部的异常光镜分析是一个由低比例（A组）向高比例（B组）转化的过程,而电镜分析没有改变。本文在A组和B组未有发现与输精管结扎术有关的精子形态学上特征性的改变。     

环孢素A转换为雷帕霉素长期作用对移植肾大鼠睾丸功能和形态学的影响

比较环孢素A(CsA)转换为雷帕霉素(Rapa)后与CsA长期作用对移植肾大鼠睾丸功能和组织形态学的影响。方法 采用标准的肾移植模型方法进行原位左肾移植,即将Fisher大鼠的供肾移植给Lewis雄性大鼠,36只大鼠分为CsA转换为Rapa组(R组,n=10)、CsA持续使用组(A组,n=10)、CsA撤离组(B组,n...     

Application of passive hemagglutination for evaluation of antisperm antibodies and a modified coombs'' test for detecting male autoimmunity to sperm antigens

A rapid, specific, and sensitive assay using passive hemagglutination and Coombs' tests has been established for detecting male autoimmunity and female isoimmunity to sperm antigens. Antisperm antibody titers evaluated by this method were similar in 156 normal females, 400 pregnant females, 25 gonadal dysgenetics, 43 users of Oracon (an oral contraceptive), and 25 normal males. In contrast, in the evaluation of 50 couples with unexplained infertility, 16 males were found to have significantly elevated Coombs' autoantibody titers (primarily IgG) and correspondingly higher antisperm antibody titers in their serum and seminal plasma; another 28 had low autoantibody titers and low serum and seminal plasma antibody titers; and 6 had intermediate titers. The wives of 63% of the males with high autoantibody titers were found to be ‘isoimmune’, as compared with 7% of those with low titers. The isoimmune females had significantly higher antisperm antibody titers than did those with normal titers. In general, comparable titer values were obtained whether sperm extract or seminal plasma was used as the erythrocyte coat for passive hemagglutination. The titers were consistently higher in the seminal plasma than in the serum of males. Very little crossreactivity was apparent between anti-Candida and antisperm antibodies, and antibodies to blood group antigens did not appear to influence the results. When the results obtained by passive hemagglutination were compared with those obtained by immunofluorescence, Franklin-Dukes, and hanging drop interface methods, only those of the passive hemagglutination and Coombs' test correlated well with the results of postcoital tests and seminal analysis.     

Effects of porcine circovirus type 2 (PCV2) maternal antibodies on experimental infection of piglets with PCV2

To determine the effects of porcine circovirus type 2 (PCV2) maternal antibodies on and response to experimental PCV2 infection, 24 piglets were divided into four groups on the basis of the enzyme-linked immunosorbent assay titers of PCV2 maternal antibodies: group A (n = 6; sample/positive [S/P] ratio, <0.2), group B (n = 5; S/P ratio, >0.2 to <0.5), and groups C (n = 8) and D (n = 5) (S/P ratio, >0.5). Piglets in groups A, B, and C were inoculated with PCV2 at day 0 and challenged with PCV2 at day 42. Group D piglets were not exposed to PCV2 at day 0 but were challenged at day 42. Before challenge, seroconversion to PCV2 antibodies occurred in five of six group A piglets, and the antibody level rose above the cutoff level in one of five group B piglets. Viremia was detected in five of six, four of five, and two of eight pigs in groups A, B, and C, respectively. After challenge, PCV2 DNA was detectable from 7 to 21 days postchallenge in the sera from six of six, four of five, three of eight, and five of five pigs in groups A, B, C, and D, respectively. The results indicated that protection against PCV2 infection conferred by maternal antibodies is titer dependent: higher titers are generally protective, but low titers are not.     

Early testicular changes after vasectomy and vasovasostomy in Lewis rats

The testes of Lewis rats were studied at intervals from 2 weeks to 3 months after bilateral vasectomy, vasectomy followed 1 month later by vasovasostomy, or sham operations. Aims were to determine the nature of early alterations after vasectomy, and to determine whether vasovasostomy after 1 month would result in reversal of vasectomy-induced changes. Approximately one-fourth of the testes in the vasectomy and vasovasostomy groups displayed histological changes, which consisted mainly of depletion of germ cells. The extent of the depletion varied greatly in different seminiferous tubules. In testes altered in this way, no abnormal infiltrations of lymphocytes, macrophages, or other cells were observed in the seminiferous epithelium or in the interstitium. The rete testis and straight tubules were normal in testes with altered seminiferous epithelium. A few testes in the vasectomy and vasovasostomy groups had necrotic centers. The results suggest that depletion of germ cells occurred as a result of shedding from the seminiferous epithelium into the lumen of the tubules. A cellular immune response, such as occurs in experimental allergic orchitis in other species, did not appear to be responsible for the observed loss of germ cells. This suggests a possible role for humoral antibody in this model, since there is an association between testicular changes and serum antisperm antibodies at longer intervals after vasectomy. Testicular alterations were not reversed by performance of a vasovasostomy 1 month after vasectomy.     

Relationship between level of serum sperm immobilizing antibody and its inhibitory effect on sperm migration through cervical mucus in immunologically infertile women

PROBLEM: Sperm immobilizing antibodies often interfere with the penetration of sperm through the cervical mucus. However, the relationship between sperm immobilizing antibody titer and the result of the post-coital test (PCT) has not yet been clarified. The aim of this study was to investigate whether the 50% sperm immobilization unit (SI50) titer, a quantitative measure of sperm immobilizing antibody, in patients' sera was correlated with the result of PCT. METHOD OF STUDY: The sperm immobilization test (SIT) was performed for 2834 infertile women in two university hospitals. The SI50 titers were evaluated by quantitative SIT. In some cases with sperm immobilizing antibody, PCT was carried out before ovulation and assessed according to the criteria of World Health Organization (1992). Infertile couples with abnormal semen characteristics were excluded. RESULTS: Seventy-four of 2834 women had sperm immobilizing antibodies in their sera, giving a positive rate of 2.6%. Twenty-four (77.4%) of 31 women with sperm immobilizing antibodies and 28 (20.4%) of 137 women without the antibody had abnormal PCT results. There was a significant difference between the two groups (P<0.0001). When patients with sperm immobilizing antibodies were divided into two groups according to the SI50 titers, the abnormal result of PCT was obtained in all 10 patients with high (>10) SI50 titers, while that was 14 (66.7%) in 21 patients with low (10) SI50 titers. There was a significant difference between the two groups (P=0.04). CONCLUSION: The SI50 titer in the serum can predict inhibitory effects on sperm migration through cervical mucus in immunologically infertile women. Evaluation of the SI50 titers in patients' sera seems to be useful for decision-making in infertile women with sperm immobilizing antibodies regarding whether they have the possibility of conceiving by timed intercourse.     

Cross-Reactivity of Sperm and T Lymphocyte Antigens*

ABSTRACT: Evidence is presented for cross-reactivity between antigens on human sperm and T lymphocytes. In 25 infertile couples in which both the males and females had significant antisperm immunity, antibody (Ab) titers to thymocytes (mean ± S.E.M. 159 ±4 and 72 ± 14, respectively, in males and females), T cell lines CCRF-CEM (69±5 and 48±8) and HSB-2 (56±15) and 41±8), suppressorenriched (T_G) cells (26±6 and 66±28) and helper-enriched (T_g—) cells (26±4 and 46±14) were significantly elevated, as compared wth Ab titers in 45 normal males and 45 normal females without antisperm immunity. Antibody titers to adult B cells, B cell line RAJI, and granulocytes were similar in the two groups. Antisperm Ab titers in sera, sperm extracts, and seminal plasma of the infertile subjects were significantly reduced after absorption with sperm, thymocytes, or T cell line CCRF-CEM but not with the B cell line RAJI. Antithymocyte Ab titers in the sera were significantly reduced (p < 0.001) after absorption with thymocytes, CCRF-CEM, or sperm, but not RAJI. Lymphocytes from the infertile patients, when stimulated with pokeweed mitogen in vitro, produced antisperm and anti-T-lymphocyte antibodies at significantly higher titers than normal controls.     

An Enzyme-Linked Immunosorbent Assay for Measuring Antisperm Autoantibodies Following Vasectomy in Lewis Rats

ABSTRACT: An indirect enzyme-linked immunosorbent assay (ELISA) was devised to measure antisperm autoantibodies in the Lewis rat following vasectomy. The assay system was validated by employing prevasectomy sera and postvasectomy antisera, previously demonstrated to contain antisperm antibodies by indirect immunofluorescence. A standardized ELISA protocol was developed employing 10⁵ sperm per microtiter plate well and sucrose-polyvinylpyrrolidone as a postcoat stabilizer solution. The ELISA was shown to yield significant detectable antibody at dilutions of 1/512 or greater in the most reactive sera. A standard for scoring positive titers was adopted: 1.96 standard deviations above the mean of the preimmune value. Using the criterion, 88% of 7-week postvasectomy samples could be discriminated from preimmune samples at a 1:16 dilution, which was adopted for subsequent assays. The ELISA demonstrated 73% and 91% reproducibility for an intraassay analysis of single prevasectomy and postvasectomy serum samples (7 weeks postvasectomy) tested in 160 determinations on a standard sperm pool. When this single antigen pool was employed in 35 determinations at 0, 1, and 4 weeks in an interassay study, 56% and 70% reproducibility was found for pre- and postvasectomy sera respectively. A correlation (r = 0.75) was made between a single absorbance value and the end-point titer of the same sera, which indicated that single absorbance values could be used to predict serum titer and single dilutions could be used for general screening of a large number of samples. The ELISA described provides a rapid, sensitive, and reliable method that discriminated between samples taken before and after vasectomy.     

Autoimmunity to sperm antigens in vasectomized men.

Sera from vasectomized men were tested for the presence of antibodies directed against sperm antigens. A high percentage (about 55%) of the vasectomized men developed agglutinating antibodies. A lower percentage (22%) also developed low titres of antibodies to human protamine, as detected in the indirect IFT on swollen sperm heads and 22% developed cytotoxic antibodies. A correlation was found between the presence of anti-protamine antibodies and the presence of agglutinating and of cytotoxic antibodies. This correlation, and also the fact that they developed after vasectomy, indicates that the formation of antibodies against human protamine is a result of an autoimmune response to spermatozoa. The indirect IFT was also performed on 'normal' unswollen spermatozoa. All the sera were positive at least on one of the sperm antigens located in the acrosome, equator, or post-nuclear region, but no increase nor decrease in titre was found after vasectomy.     

柯萨奇病毒B组3型亚单位疫苗与基因靶向疫苗免疫小鼠的效果比较

目的:观察柯萨奇病毒VP1基因靶向核酸疫苗pcDNA3/C3d3-sVP1、重组腺病毒rAd/C3d3-sVP1和亚单位疫苗VP1蛋白的免疫效果。方法:雄性BALB/c小鼠随机分为4组,每组18只,分别肌肉注射PBS、pcDNA3/C3d3-sVP1、rAd/C3d3-sVP1和VP1蛋白,除重组腺病毒rAd/C3d3-sVP1组免疫2次,间隔2周外,其余3组均免疫3次,间隔3周。质粒每次每只接种100μg/100μl,腺病毒每次每只接种1.2×107pfu/100μl,蛋白每次每只接种50μg。分别用ELISA法和微量中和试验法检测血清CVB3 VP1特异性IgG抗体和中和抗体;CCK-8法检测脾淋巴细胞特异性CTL杀伤活性;以致死量的CVB3病毒液感染已免疫小鼠,检测小鼠血中病毒滴度,观察存活情况以评价各种疫苗的免疫保护作用。结果:各实验组小鼠的特异性IgG抗体和中和抗体滴度随免疫次数增加而提高,末次免疫后,VP1蛋白组的特异性IgG抗体和中和抗体滴度均明显高于pcDNA3/C3d3-sVP1组和rAd/C3d3-sVP1组(P<0.05),但rAd/C3d3-sVP1组CTL杀伤活性高于pcDNA3/C3d3-sVP1组和VP1蛋白组;经致死量CVB3感染后,VP1蛋白组血中病毒滴度低于其他实验组,而生存率明显高于其他各组。结论:VP1蛋白疫苗诱导小鼠产生较强的特异性免疫应答,提高了小鼠的生存率,免疫效果优于靶向基因疫苗pcDNA3/C3d3-sVP1和rAd/C3d3-sVP1。     

Changes in antibody titers to hepatitis C virus following interferon therapy for chronic infection

The use of quantitative assays for hepatitis C virus specific antibodies (anti-HCV) as a prognostic marker was evaluated in 31 patients with chronic hepatitis C treated with interferon (IFN). Changes in titers of serum HCV-RNA and anti-HCV antibodies; anti-C11 (anti-core), anti-C100 (anti-NS3), and anti-C7 (anti-NS3) were investigated. Recombinant IFN-α 2a was administered and the patients were followed for more than 1 year. The patients were classified into three groups according to their responses to IFN: 11 sustained responders with continuous normalizations of serum alanine aminotransferase (ALT) levels; 14 transient responders with transient decreases in ALT; and six nonresponders who had no changes in ALT levels. Ten of 11 sustained responders had a continuous decrease in anti-C11 titers after completion of treatment, decreasing to less than half of pretreatment titers. No patients in the other two groups had a continuous decrease in anti-C11 titers. Although sustained responders had decreases in anti-C100 and anti-C7 titers after IFN therapy, these titers also decreased in some patients in the other two groups. HCV-RNA was not detected in the sera of 10 of 11 sustained responders following IFN therapy. In contrast, while 9 of 10 transient and nonresponders had a decrease or disappearance of HCV-RNA at the completion of therapy, they had increased levels thereafter. These results indicate that anti-HCV-core (anti-C11) titers most closely reflect the status of HCV replication. A quantitative assay for anti-HCV-core antibody can be used as a predictive marker of remission in IFN-treated patients with chronic hepatitis C. © 1994 Wiley-Liss, Inc.     

输精管结扎家兔自身免疫反应的实验研究

33只成年雄兔行双侧输精管结扎手术后,进行连续12个月的免疫学观察。7只同龄雄兔作为空白对照。结果表明,间接血凝试验69.7%结扎组家兔检测出抗精子抗体,滴度范围为1:5～1:1280;间接免疫荧光测定有90.9%的实验组家兔测出抗精子抗体。白细胞粘附抑制实验呈阳性反应者占实验组家兔的43.5%,与对照组比较差异显著。应用PEG光密度和抗补体法测定CIC均里阴性。输精管结扎后第3个月附睾肿胀者占45%,第5—7个月达70%,之后逐渐消退。而与此同时抗精子抗体阳性检出率明显增高,故可推断精子抗原主要经附睾入血,作用于免疫系统导致体液与细胞免疫反应。结扎组家兔胸腺、脾、淋巴结呈明显的增生现象支持这一结论。     

Effects of active and passive immunization in the Mycoplasma arthritidis infection of rats

Viable, heat-inactivated, formalin-inactivated and sonicated Mycoplasma (M.) arthritidis antigens as well as immune sera against M. arthritidis and spleen cells from Lewis rats recovered from M. arthritidis infection were injected into naive Lewis rats prior to a challenge infection with 10(7) cfu of M. arthritidis and tested for their protective effects. Viable mycoplasmas induced arthritis combined with the production of high titers of antibodies against M. arthritidis and resistance to a second infection. The application of the inactivated M. arthritidis-antigens in emulsion of incomplete Freund's Adjuvant (ICFA) to naive rats, which also induced a strong antibody production, as well as the inoculation of reconvalescent serum from rats infected with M. arthritidis and hyperimmune serum against M. arthritidis from rabbits and mice protected rats also from an outbreak of arthritis after challenge. The injection of sonicated M. arthritidis antigen without ICFA which failed to induce the production of antibodies and the transfer of spleen cells from rats recovered from M. arthritidis infection had no protective function. The investigations showed, that antibodies play an important role in the prevention of M. arthritidis infections in Lewis rats.     

Allergic infertility: laboratory techniques to detect antispermatozoal antibodies

Twenty sera from fertile and infertile women were examined for antibodies to spermatozoa or to seminal plasma, to determine which of the five techniques tested provided reliable evidence of allergic infertility. Ten sera agglutinated seminal plasma coated tanned red cells, six agglutinated spermatozoa, ten gave mixed cell antiglobulin agglutination and fourteen gave immunofluorescent reactions for IgG or igM antispermatozoal antibodies, and three were cytotoxic to sperm. The results of tests for seminal plasma agglutinins were unrelated to those of any of the other tests, and it is believed that factors other than antibody also clump these antigen-coated cells. There was good agreement in the results obtained by direct sperm agglutination, mixed cell antiglobulin agglutination and strong immunofluorescent staining of the sperm head. The antiglobulin and immunofluorescent techniques also detect nonagglutinating (‘incomplete’ or ‘univalent’) antibody. One of the three sera cytotoxic to sperm had no antibody detected by the other tests. The sperm may have been killed by an inflammatory substance or a systemically absorbed contraceptive. This case illustrates the necessity of combining tests for cytoxicity with at least one technique for antispermatozoal antibody in order to detect allergic infertility.     

The Immune Response to Vasectomy and its Relation to the HLA System

We examined 188 men who had undergone vasectomy up to 6 years previously and, for comparative purposes, 100 men who were about to undergo the operation. Blood specimens were available from a total of 283 men. Sperm antibody assays using immunofluorescence, microagglutination, and microimmobilization confirmed that the prevalance of several types of antibody is higher after vasectomy. Immobilizing and agglutinating antibodies appeared to be the most important. Trends in antibody prevalence according to the time after the operation were analyzed. Screening against lymphocytes and lymphoid lines showed that the anti-sperm activity of era was not related to anti-HLA or anti-Ia activity. Associations were examined between different types of sperm antibody, and between these antibodies and autoantibodies to other antigens. When antibody prevalence was studied in relation to HLA types, the HLA antigen A28 was found to be strongly associated with production of head agglutinating antibody (and immobilizing and immunofluorescent equatorial antibodies) after vasectomy. This is one of the first clear-cut examples of antibody production associated with the HLA system.     

Chronic Proliferative Arthritis of Mice Induced by Mycoplasma arthritidis II. Serological Responses of the Host and Effect of Vaccines

Complement-fixing antibodies were first detected in mice 7 days after intravenous injection with Mycoplasma arthritidis. Peak titers were observed at 21 days, and high levels of antibody persisted through 293 days. The metabolism-inhibiting antibody response was minimal. On fractionation of mouse sera, only 7S antibody was detected which first appeared at 12 days after injection and persisted throughout the experiment. In contrast, serum taken from rats injected with M. arthritidis contained predominantly 19S antibodies in the early stages of the disease which were gradually replaced with 7S antibodies. The intravenous injection of mice with M. arthritidis culture supernatant fluid had no effect upon their subsequent susceptibility to the arthritogenic effects of M. arthritidis, but this procedure appeared to delay the onset of abscess formation after the subcutaneous injection of M. arthritidis. Formalin-killed cells of M. arthritidis partially protected mice against the arthritis induced by M. arthritidis. Previous infections with M. arthritidis conferred partial immunity against the arthritogenic effects of the organism. Serum taken from convalescent mice at 41 days had a partial protective effect when used to immunize passively normal mice against M. arthritidis. However, rabbit anti-M. arthritidis serum which possessed higher complement-fixing and metabolism-inhibiting antibodies was without significant protective properties.     

Pathological and Immunological Effects of Surgically Induced Varicocele in Juvenile and Adult Rats

ABSTRACT: The presence of a varicocele in adult men has been correlated with infertility. This study documents the effect of an experimentally induced unilateral varicocele in 21-day-old juvenile prepubertal and 51-day-old adult rats (n=10 per group) on subsequent adult testicular function. Varicoceles were induced by partial occlusion of the spermatic vein. There were ten sham-operated and five nonoperated control rats in each age group. The rats were sacrificed 1 month after surgery. Intrascrotal temperatures were elevated in both groups with varicoceles. Histologically, the ipsilateral testes of rats in both age groups demonstrated a decrease in the numbers of functioning seminiferous tubules and germ cells, but the decrease was significantly greater in the juveniles than in the adult rats. No changes were seen in the contralateral testes. Significant titers of cytotoxic sperm antibodies were present in all animals with varicoceles, which is in contrast to controls. The juveniles had significantly lower antibody titers (mean log ± SEM; 3.2 ± 0.09 vs. 8.5 ± 1.1, P < 0.001) than the adults. The induction of a unilateral varicocele damaged spermatogenesis and testicular function to a greater extent in juveniles than in adult rats. This damage may be immune complex-mediated.