Conception rates in couples where autoimmunity to sperm is detected

In men with autoimmunity to sperm, a varying proportion of spermatozoa in the ejaculate are found to have surface-bound immunoglobulins. We asked whether the extent of autoimmunity, as judged by this criterion, would have predictive value in determining the chance of conception. Infertile couples where husbands were found to have antisperm antibodies were treated for other factors leading to impaired reproduction, but no specific treatment was offered for reduction of these antibodies. The chance of pregnancy for those couples where autoimmunity to sperm was the sole definable factor leading to infertility was 15.3% when most spermatozoa were antibody-bound. A significantly greater number of wives whose husbands had less than 50% of their sperm bound by immunoglobulins conceived (66.7%; P less than 0.005).     

Circulating antisperm antibodies in recurrently aborting women

One hundred seventy-three women with a history of three or more recurrent consecutive abortions were analyzed for circulating antisperm antibodies with a radiolabeled antiglobulin assay (RAA), a modified enzyme-linked immunosorbent assay (ELISA), a tray agglutination test (TAT), and a sperm immobilization test (SIT). No pregnancies were subsequently gestated to term in women who were antisperm antibody-positive unless they were inoculated with their husband's leukocytes as treatment for an immune basis (not related to antisperm antibodies) for their recurrent abortions. In women with an immune basis for their recurrent abortions, immunization with leukocytes from their male partners increased the ability of these women previously aborting their fetuses to carry their fetuses to term, even if they had positive results in the ELISA, TAT, and SIT; women with positive results in the RAA continued to abort subsequent pregnancies, despite leukocyte immunization. Immunization of antisperm antibody-positive women with their partner's leukocytes did not incite or increase the antisperm antibody titer, with any of the assay techniques.     

Seminal immune response in infertile men with leukocytospermia: effect on antioxidant activity.

OBJECTIVE: To investigate the incidence of leukocytospermia and relation to T helper cytokines, tumor necrosis factor alpha (TNFalpha) and interleukin-4 (IL-4), antisperm antibodies and antioxidant activity. Design: Semen samples from 176 infertile men and 24 fertile controls were investigated. METHODOLOGY: The protocol included tubal patency test, hysterosalpingography and laparoscopy and dye test and ovulation through mid-luteal phase progesterone for the wives. The husbands had semen analysis, cytomorphology evaluation and semen culture. Seminal TNFalpha and IL-4, antisperm antibodies, total antioxidant activity, superoxide dismutase and zinc were determined. RESULTS: Leukocytospermia occurred in 44.3% of the infertile men compared to 12.5% of the fertile men (P<0.01). Thirty-six (20.5%) men had pathogenic bacterial organisms which constituted 46.2% of those with leukocytospermia. Sperm parameters were worse with leukocytospermia in terms of sperm count (P<0.01), total motility progressive motility (P<0.01), morphology, asthenozoospermia, sperm membrane integrity and antisperm antibodies. TNFalpha and IL-4 had an inverse relationship; the expression of TNFalpha was higher with leukocytospermia and bacteriospermia (P<0.001), while IL-4 was higher in fertile controls (P<0.005). Incidence of antisperm antibodies was higher with leukocytospermia. Total antioxidant activity, superoxide dismutase and zinc were lower with leukocytospermia. CONCLUSION: Leukocytospermia impairs sperm function through reduced antioxidant activity and enhanced T helper 1 modulation.     

ELISA for antisperm antibodies in unexplained infertile women

It is commonly believed that autoimmune or isoimmune responses to human sperm antigens are associated with human infertility. We examined an enzyme linked immunosorbent assay (ELISA) kit used in the detection of antisperm antibodies in the sera of women with unexplained infertility. 1) For the ELISA assay, an absorbance at 405nm greater than 0.300 was considered positive. Of the 83 sera of infertile women studied, 20(24.1%) were positive. This value was significantly higher than that of the control group: 10% of pregnant women, 11.1% of unmarried women and 10% of healthy men (p less than 0.05). 2) The results obtained with ELISA were compared with those of the sperm immobilization test. Comparison of the results of ELISA with those of the sperm immobilization test indicated that these methods detect a different, though often overlapping, spectrum of antibody activity. 3) The results obtained with ELISA did not always correlate with those of the Huhner test. However the ELISA method may be useful in the clinical screening of antisperm antibodies because of its simplicity and rapidity.     

用ELISA法检测不孕夫妇抗精子抗体——附137对观察结果

本文应用ELISA检测不明原因不孕夫妇血清抗精子抗体,结果抗体阳性率为35%,其中不孕女方阳性21.9%,不育男方阳性13.1%。而FD法和SIT法抗精子抗体阳性率各为28.5%与21.9%,三法总符合率为90.2%。不孕组抗精子抗体阳性与对照组比较,有显著统计学意义(P<0.01)。显示不孕夫妇抗精子抗体的发生明显高于生育组。本文就抗精子抗体的发生与不孕症的关系、诊断及其临床意义进行了讨论。     

Detection and titration of class-specific antisperm antibodies in serum using an enzyme-linked immunosorbent assay

An antisperm antibody enzyme-linked immunosorbent assay (ELISA) that uses whole unfixed sperm and detects immunoglobulin G (IgG) and IgA antibodies in serum was developed. Donor sperm were washed and plated on poly-L-lysine-treated microtiter plates. The patient's sera were diluted to concentrations of 1:4 to 1:256 and incubated with sperm. Positive and negative sera had been previously tested for IgG antisperm antibody activity with a radiolabeled antiglobulin assay. Samples were considered positive when the mean absorbance of triplicate wells was greater than 2 SD above the pooled negative mean. Intra-assay variation was 7.9 and 9.6% for pooled negative and positive controls, respectively. Identical titers of control positive serum were consistently detected. A correlation of 0.83 was observed between ELISA IgG serum titers and radiolabeled antiglobulin results (N = 12). All negative samples tested negative in both assays (N = 21). Some serum samples showed IgA antisperm antibodies. Determination and titration of class-specific antibodies in serum should facilitate initial screening and follow-up of patients at risk for antisperm antibodies.     

Antispermatozoal antibodies in men with urethritis

Antispermatozoal antibodies have been measured by the tray agglutination test (TAT) in three groups of patients with urethritis attending a clinic for sexually transmitted diseases and in one control group of men without urethritis. In 3 of 17 (17.6%) patients with acute nongonococcal urethritis (NGU) the serum TAT was positive at titers of 1:16 or more; and in 1 of these patients in whom Chlamydia was grown, the TAT titer rose from 1:4 before treatment, to 1:8 at 2 weeks, and 1:16 at 4 weeks, indicating probable immunization against sperm antigens at the time of infection. A rise in titer from 0 to 1:8 occurred in a second patient with NGU. Six (15.6%) of 39 patients with recurrent NGU and 2 (16.6%) of 12 patients with gonorrhea followed by postgonococcal urethritis also had positive antisperm antibody titers of 1:16 or more. None of 27 control subjects had positive antisperm antibody titers. These observations indicate that development of antisperm antibodies can be stimulated in some individuals by NGU.     

Sperm antibodies and human in vitro fertilization

In order to directly evaluate the effects of sperm antibodies in human in vitro fertilization (IVF), the authors preincubated donor sperm in female sera containing sperm antibodies and then inseminated supernumerary human oocytes from a gamete intrafallopian transfer (GIFT) program. The sperm were incubated for 30 minutes in medium containing 20% serum with antisperm activity (Test); or no antisperm activity (Control) as assessed by the immunobead test (IBT). Each oocyte was inseminated with 1 to 2 X 10(5)/ml of the preincubated motile sperm with Control or Test treatments allocated on a random basis. Six positive sera were tested in 17 experiments, resulting in a fertilization rate of 41% (25/61) versus 84% (36/43) for controls (P less than 0.001). When considered individually, three of six positive sera caused significant inhibition. The only serum that gave complete inhibition had the highest titer for IgG (10,000) and lower IgA (100). Absorption with protein A reduced the IgG titer to less than 10 and removed the fertilization inhibitory activity. These results confirm that sperm antibodies from female sera can inhibit human IVF.     

溶脲脲原体感染与血清中抗精子抗体相关性研究

本文对７１例不育男性进行了精液ＵＵ培养及血清中抗精子抗体（ＡＳＡｂ）的检测（ＥＬＩＳＡ法）。结果可见，精液培养ＵＵ阳性者３８人，其中血清ＡｓＡｂ亦阳性者１０人，占２６．３２％；精液培养ＵＵ阴性者３３人，其中血清抗精子抗体阳性者２人，占６．０６％；两者相比，差别显著（Ｐ＜０．０５）。对不孕女性４５例也进行了同样试验，阴道分泌物ＵＵ培养阳性者２２人，其中血清ＡｓＡｂ阳性者９人，占４０．９１％；阴道分泌物ＵＵ培养阴性者２３人，其中血清中ＡｓＡｂ阳性者２人，占８．７０％，差别显著（Ｐ＜０．０５）。实验结果表明男女不育者ＵＵ感染与血清中ＡｓＡｂ的增高有一定的关系。     

Effects on fertility of immunizing mice with anti-idiotypic antibodies to porcine zona pellucida antigen

OBJECTIVE: To evaluate the effects on fertility by immunization with anti-idiotypic antibodies to porcine zona pellucida (PZP) antigen. METHOD: Anti-idiotypic antibodies (Ab(2)) were produced in New Zealand rabbits immunized with 17D3 monoclonal antibodies (mAbs) (IgG, Ab(1)) to PZP antigen. The antisera were first passed through immuno-affinity chromatography column linked to normal mouse IgG so as to remove the antibody bound to normal mouse IgG The passing elute was then purified by immuno-affinity chromatography using 17D3 mAbs to get the Ab(2). Female BALB/c mice, 5-week-old, were grouped and immunized with the Ab(2), PZP antigen, target antigen of the Ab(1) and normal rabbit IgG, respectively. The treated female mice were mated with male BALB/c mice and sacrificed at gestation day 10. Analyses included ELISA measurement of anti-ZP antibody titer, fetal number determination and evaluation of ovarian histomorphology. RESULTS: The Ab(2) appeared as a single protein band by SDS-PAGE. Shown by a competitive inhibition ELISA, the Ab(2) specifically bound to the variable region of the 17D3 Ab(1). Compared with controls, the female mice immunized with Ab(2) showed a decreased pregnancy rate and a statistically significant reduction in fetal numbers. Histological examination of ovaries demonstrated that Ab(2) exposure interfered less with follicular development than did exposure to PZP. CONCLUSION: Immunization of female mice with Ab(2) to PZP mAbs suppresses fertility and fetal numbers with minimal ovarian pathology.     

无精子症配偶宫颈炎状况及其对宫颈黏液影响的临床研究

目的:探讨无精子症配偶宫颈炎状况及宫颈炎对宫颈黏液的影响。方法:对在我院助孕的无精子症配偶500例,取阴道分泌物及宫颈管黏液行病原体检查、白细胞检测;同时抽静脉血行ELISA法抗精子抗体(AsAb)、单纯疱疹病毒(HSV)、巨细胞病毒(CMV)检测。围排卵期进行宫颈黏液评分,血AsAb升高者进一步行围排卵期宫颈黏液毛细管测试及精子-宫颈黏液接触试验。结果:①按照诊断标准,500例妇女分为宫颈炎组92例(18.4%),对照组408例(81.6%)。②宫颈炎组宫颈黏液CT、UU、BV、HPV感染率较对照组明显增高,差异均有高度统计学意义(P<0.01)。宫颈炎组血HSV、CMV感染率较对照组明显增高,差异均有统计学意义(P<0.05)。③宫颈炎组围排卵期宫颈黏液评分较对照组(9.32±1.91分对12.67±2.14分)增加,差异有统计学意义(P<0.05)。两组宫颈黏液的量、成丝性、羊齿化结晶程度比较差异无统计学意义(P>0.05);而宫颈炎组较对照组宫颈黏液黏稠、黏液细胞数明显增多,差异均有高度统计学意义(P<0.01)。④宫颈炎组血AsAb阳性检出率33.7%(31/92),明显高于对照组9.8%(40/4...     

Effect of sperm antibodies on pregnancy outcome in a subfertile population

The relationship between sperm antibodies, conception, and miscarriage was examined in 109 infertile couples. Antibodies present on the surface of husbands' ejaculated sperm and antibodies in husbands' or wives' sera that reacted with a purified population of the husbands' motile spermatozoa were detected by an enzyme-linked immunosorbent assay. During an 18-month period, conception occurred in 33 (30.3%) of the couples; 16 (14.7%) women subsequently suffered a spontaneous miscarriage during the first trimester, whereas 17 (15.6%) women maintained their pregnancies past this time period. Antisperm antibodies were present in sera from only two of 17 (11.8%) women with successful pregnancies, whereas seven of 16 (43.8%) women who miscarried and 29 of 76 (38.2%) who did not conceive had these antibodies in their sera. IgG (22.4%) and IgM (21.1%) antisperm antibodies predominated in sera of women who did not conceive, whereas IgA (37.5%) and IgG (37.5%) antibodies were most prevalent in sera of women with miscarriages. In men, the presence of antisperm antibodies in sera was unrelated to fertility. However, there was a correlation between sperm surface antibodies and an inability to conceive. IgG was identified on ejaculated spermatozoa from eight of 76 (10.5%) men whose wives failed to conceive and in none of 33 men whose wives conceived. Similarly, IgA was present on spermatozoa from 16 (21.1%) infertile and two (6.1%) fertile men. Thus antisperm antibodies in female sera and on ejaculated spermatozoa were associated with a failure to conceive and first-trimester miscarriage.     

Relationship between circulating antisperm antibodies in women and autoantibodies on the ejaculated sperm of their partners

The relationship between antibodies on the surface of ejaculated sperm and circulating antibodies in female partners was evaluated. Of 616 couples examined by the immunobead binding test, there was a 12.4% incidence of sperm-surface antibodies in men whose wives had antisperm antibodies in their sera, but only a 6.5% incidence in partners of women who lacked these antibodies (p less than 0.025). Sperm-bound immunoglobulin G and immunoglobulin A both occurred at a significantly higher frequency (p less than 0.05) in partners of women with serum antisperm antibodies. Increased incidence of both immunoglobulin G (p less than 0.01) and immunoglobulin M (p less than 0.005) circulating antisperm antibodies in females were observed when the male partners had antibody-bound sperm. Antibody-coated sperm may activate lymphocytes in the female partners after coitus, thus leading to the production of antisperm antibodies. This may be an additional mechanism that leads to female isoimmunity to sperm and infertility.     

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.     

Antisperm antibodies and in vitro fertilization failure

A new enzyme-linked immunosorbent assay (ELISA)-based test (Zer, Jerusalem) has allowed us to show the presence of antisperm antibodies (1:32 to 1:64) in the blood of 14 (33%) of 32 patients undergoing in vitro fertilization and embryo transfer (IVF-ET) under gonadotropin stimulation. Observation of the morphology of fertilization in eight patients with and seven patients without antisperm antibodies has shown a significant association (P 0.025. Fisher exact probability test) among the cumulus/corona coagulation, the absence of fertilization, and the presence of these antibodies. Cumulusloocyte complex washing and/or enzymatic cumulus removal are considered as elective interventions in the case of antisperm immunity. Each patient entering an IVF-ET program should have the antisperm antibody assay performed as a preliminary screening.     

Sperm antibodies and human leukocyte antigens in couples with early spontaneous abortions

Twenty-six couples with two or more early spontaneous abortions and 53 couples with recently proven fertility (control group) were typed for human leukocyte antigens (HLA) of A and B loci and screened for the presence of cytotoxic and hemagglutinating antibodies to sperm. Eighteen other couples with two or more abortions were tested for sperm antibodies alone. Both partners in 24 of 44 (54%) aborting couples were positive (titers of greater than or equal to 32), whereas the control group had sperm antibodies in the negative range (titers of 0 to 16; P less than .0001). Thirty-one husbands (70%) and 24 wives (54%) were positive for sperm antibodies in the study group. Sharing of HLA-A and/or -B antigens between partners was similar in the control and study groups. Fourteen of 26 (54%) couples with two or more abortions did not share HLA-A and/or -B antigens, in contrast to 14 of 53 (26%) in the control fertile group (P less than .02). Antigenic frequencies of HLA-B7 singly and in combination with HLA-B35 were increased in females in the aborting but not in the fertile control group (P less than .001). The combined incidence of HLA-B7, -B8, and -B35 in both partners was significantly higher in the aborting couples as contrasted with the control group. It is suggested that the presence of sperm antibodies is associated with early pregnancy wastage. Histocompatibility antigens B7 and B35 may play a role through their association with sperm antibodies and early spontaneous abortions.     

ELISA法检测不育男子精浆中抗精子IgG和IgA

将精子经Tritonx－100处理，冷冻高速离心后，经抗人全血清-SephadexG－75亲和柱层析分离，提取分子量为59KD人精子膜蛋白作为抗原，经ELISA间接法对20例生育男子和50例不育男子精浆中抗精子IgG和IgA进行了测定。结果显示：生育男子精浆中抗精子IgG为阴性；抗精子IgA阳性率为5％。不育男子精浆中抗精子IgG和IgA阳性率分别为10％、30％；抗精子IgG和IgA均为阳性者3例，均为阴性者33例；抗精子IgG阳性而IgA阴性者2例；抗精子IgA阳性而IgG阴性者12例。不育组与生育组间抗精子IgG阳性率无显著性差异（P＞0．05）；而IgA阳性率间则有显著性差异（P＜0．05）。不育组抗精子IgG和IgA阳性率间有极显著性差异（P＜0．01）。本文利用ELISA法对精浆中抗精子抗体的分类及可能来源进行了讨论。     

Differences between husbands' and wives' approach to infertility affect marital communication and adjustment

Objective: To test a theoretical model of the effect on marital communication and adjustment of men’s and women’s approach to infertility.

Design: A cross-sectional research design involving interviews, questionnaires, and a marital discussion task.

Setting: Volunteers from practices of fertility specialists.

Patient(s): Forty-eight couples currently seeking infertility treatment.

Intervention(s): None.

Main Outcome Measure(s): Quality of marital communication during a marital discussion task and effect of infertility on the marriage.

Result(s): Having children was more important to wives than husbands; wives were more involved in trying to have a baby, wanted to talk with their partner more about trying to have a baby, and experienced a greater loss of self-esteem than did their husbands. To the extent that husbands saw having children as important, were involved in trying to have a baby, or wanted to talk with their wives about trying to have a baby, the quality of marital communication when discussing infertility was less negative, and in turn, wives perceived a more positive effect of infertility on their marriage.

Conclusion(s): Increases in husbands’ interest and involvement in fertility treatment may lead to positive changes in couple communication about infertility and to a more positive effect of infertility on the marriage.     

An ELISA for antisperm antibody detection in serum: comparison with TAT and SIT in serum, with MAR-test, immunobead-test and TAT in semen and with micro-SIT in cervical mucus

16 couples belonging to couples with negative or doubtful PCT were selected according to the presence of antisperm immunization. 12 patients, 5 male and 7 female, showed both localized and generalized immunization. The former was diagnosed by means of a positive IgG MAR-Test, direct IgG Immunobead-Test, direct IgG Immunobead-Test and seminal TAT in the male patients, and Micro-SIT in the cervical mucus of the female patients, while for the latter there was simultaneous positivity of both serum TAT and SIT, except for two cases, in which the SIT only was positive. The 4 remaining patients, 2 male and 2 female, did not show any signs of antisperm immunization. The evaluation of the antisperm antibodies by means of the ZER ELISA Antisperm Kit in the serum of the 16 patients examined showed that there were no significant statistical differences between the serum TAT and the SIT. The former showed agreement of the results in 93.75% of the cases, and the latter in 81.25%. A strict correlation was observed between the ELISA for serum antisperm antibodies (ELISA-AS-Abs) and the local immunitary situation, with agreement in 93.75% of the cases. The ELISA-AS-Abs seems to bring the advantage of eliminating the need for fresh semen for antibody titration and also means that there is no subjective interference with the evaluation of the results.     

Mexican women's reasons for accepting or declining HIV antibody testing in pregnancy

OBJECTIVE: To determine pregnant women's reasons for accepting or declining the HIV test in Leon, Mexico. DESIGN: A cross-sectional study using a face-to-face questionnaire. SETTING: The antenatal clinic at a tertiary-care referral hospital in Leon, Mexico. PARTICIPANTS: 1184 pregnant women. DATA COLLECTION: Reasons for accepting or declining the HIV antibodies test, socio-economic characteristics and risk factors for HIV were recorded. Blood samples were obtained from women who accepted to be tested, and positive serologies to HIV on duplicate enzyme-linked immunosorbent assay testing were confirmed by Western Blot assay. FINDINGS: 1009 (85.2%) women accepted the HIV antibodies test. The main reason for accepting it was that women felt the test could be beneficial to their babies (45.1%). The two main reasons for rejecting the HIV antibodies test were that women felt the test was unnecessary because their husbands did not have sexual intercourse with other women (32.6%), and because they did not have permission from their husbands for accepting the test (23.5%). None of the women tested positive for HIV antibodies (0 per 1009). KEY CONCLUSIONS: The reasons for accepting the HIV test were similar to those reported in developed countries. One important reason for declining the test was that women did not have their husband's permission. IMPLICATIONS FOR PRACTICE: The acceptance rate for HIV testing in pregnant women could be improved by counselling men on the value of their wives being tested in pregnancy.