Chlamydial serology in infertile women by immunofluorescence.

The chlamydial serum immunoglobulin G antibodies of 128 infertile women were determined by a single-antigen (L2) immunofluorescence test. The antibody titers were compared with those of female contacts of men with nonspecific urethritis and with those of "normal" pregnant women. Among infertile women, 43% lacked chlamydial antibodies (titer less than or equal to 8). The corresponding proportion for contacts of men with nonspecific urethritis was 14% and that for pregnant women 71%. A very high titer (greater than or equal to 256) was recorded in 25 infertile women and in 7% of pregnant women. Among women with positive chlamydial isolation, 46% had titers greater than or equal to 256. Bilateral tubal obstruction was demonstrated in 23 infertile women (18%). The geometric mean titer of these patients was higher than that of patients with normal hysterosalpingograms. In nine cases with bilateral tubal obstruction and very high chlamydial antibody titers, there was no history of pelvis infections. The operative findings were typical of previous salpingo-oophoritis.     

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.     

Clinical study on circulating antisperm antibodies in women with recurrent abortion

Couples with a history of recurrent spontaneous abortions (RSA) were analyzed for circulating antisperm antibodies (Ab) with an enzyme-linked immunosorbent assay (ELISA), a Franklin-Dukes (F-D) and a sperm immobilization test(SIT). The three tests were positive in 31.5%, 25.9% and 18.5% for wives and 24.1%, 11.1% and 7.4% for husbands in RSA couples. There was a statistical significance (P less than 0.01) as compared with the control group. The ELISA method was found to be most sensitive, and the positive rate of antisperm antibodies in wives was higher than that in husbands. The Ab titer of RSA group varied within 1:8 approximately 1:512, with 60% above 1:32. The pregnant weeks (P greater than 0.05) did not seem to affect the antisperm Ab titer, nor did the aborting frequency. Our study suggests that antisperm Ab is one of the important causes of RSA in women.     

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.     

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.     

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.     

The significance of serum specific IgA antibody to Chlamydia trachomatis in the diagnosis and treatment of chlamydial infection in female genital tract

Chlamydia trachomatis IgA and IgG antibody titers were determined by indirect immunoperoxidase assay in sera of 88 female patients with C. trachomatis-positive cervicitis and 55 C. trachomatis negative healthy controls. Serial serum samples obtained until 66 weeks after treatment were also studied for IgA and IgG antibodies in 28 of the 88 patients with chlamydial cervicitis. In the 88 patients, the presence (94.3%) of antibody (titer greater than or equal to 16) was comparable to that (100%) of IgG antibody (titer greater than or equal to 32), but in healthy controls the presence (5.4%) of IgA antibody was lower than that (30.6%) of IgG antibody. Serum IgA antibody decreased gradually and was undetectable (titer less than or equal to 8) at 20 weeks after treatment in 78.6% of the 28 patients. Serum IgG antibody persisted at stable levels within the same period. These data suggest that the presence of serum IgA antibody to C. trachomatis is correlated with active C. trachomatis infection of the female genital tract.     

Occurrence of serum antisperm antibodies in patients with cystic fibrosis

OBJECTIVE: To determine if acquired obstruction of the vas deferens in men with cystic fibrosis (CF) induced the development of antisperm antibodies with genital tract obstruction similar to other men. DESIGN: Serum antisperm antibodies were assayed by an indirect immunobead test and an indirect immunofluorescence assay. Both homologous (human sperm/human zona) and heterologous (human sperm/zona-free hamster ova) sperm/egg interactions were evaluated in the presence of serum antisperm antibodies from patients with CF. SETTING: Cystic Fibrosis Clinic at the University of Oklahoma Health Sciences Center, a tertiary care referral center. PATIENTS: Fifteen CF patients (10 male and 5 female), 3 non-CF antisperm antibody-positive infertile patients (2 male and 1 female), 20 fertile controls (7 males and 13 females), and 9 fertile sperm donors were used. INTERVENTIONS: None. MAIN OUTCOME MEASURES: Serum antisperm antibody levels in patients with CF. In those patients with antisperm antibodies, determine effect of these sperm antibodies on sperm/egg interactions and complement-mediated events. RESULTS: Sera from 3 (30%) of 10 men with CF demonstrated immunoglobulin (Ig)G, IgA, and/or IgM antisperm antibodies, whereas sera from all 5 CF women and the 20 control sera were negative for antisperm antibodies. The maximal titers for IgG, IgA, and IgM antisperm antibody were 1:8, 192, 1:256, and 1:64, respectively. The immunobead binding, which was restricted to the sperm head and tail-tip or the midpiece and tail-tip, correlated with the indirect immunofluorescence pattern. Antisperm antibody-positive sera from men with CF impaired both the binding and penetration of human zonae and the penetration of hamster ova by human sperm. CONCLUSIONS: Similar to other men with congenital or acquired obstruction of their genital tract, antisperm antibodies may occur in some men with CF. Antisperm antibodies may contribute to immune sperm dysfunction in some men with CF by activated complement-mediated events and interfering with sperm/egg interactions.     

Qualitative differences in sperm antibody responses in mice of different inbred strains and sexes

To better understand the immunogenetic basis and potential pathological consequences of anti-sperm humoral immunity, age-matched female mice of 9 different inbred strains were immunized with syngeneic sperm and were tested for qualitative (specificity) and quantitative (titer) antibody differences by radioimmunoassay, immunofluorescence and immunoblot techniques. All mice developed antisperm antibodies, although titers varied considerably between inbred strains. Antisperm antibodies produced in this study did not cross-react with membrane antigens on thymocytes, brain or immature testicular germ cells. Immunoblot tests identified 17 major sperm antigen bands; this approach also revealed considerable inter- and intra-strain variation in antisperm antibody specificities among female mice. In a parallel study C57BL/6 male mice demonstrated significantly lower antisperm antibody titers and an absence of response to certain sperm antigens in immunoblot tests when compared to age-matched females of the same inbred strain. These findings provide evidence that genetic factors (including sex) interact with environmental (nongenetic) factors in the control of immune responses to sperm antigens.     

Post partum rubella vaccination and anti-D prevention]

The effectiveness of a routinely performed puerperal rubella vaccination was tested. Additional a possible adverse influence of simultaneously administered anti-D immunoglobulin on the effectiveness of the rubella vaccination was examined. Rubella antibody titers (HHT) in pregnant women were determined; after delivery puerperal women with titers of less or equal 1:16 were selected for rubella vaccination. 2 1/2 to 3 months later rubella antibody titers were done again. 15% of 130 vaccinated women did not show a conversion of the former negative titer or a low titer of 1:8 remained. Also reductions of the antibody titers were seen. When simultaneously rubella vaccination and anti-D immunoglobulin was administered only in 1 case out of 27 patients a negative titer remained after vaccination. The used anti-D immunoglobulin contained rubella antibodies of a titer 1:256 to 1:512; according to experimental studies, this concentration should not have any influence to the immunologic response of rubella vaccination. Our practic results could not confirm the reservations concerning simultaneous rubella vaccination and anti-D prophylaxis.     

Antisperm antibody binding to human acrosin: a study of patients with unexplained infertility

OBJECTIVE: Antisperm antibody binding to acrosin was investigated by Western Blotting. The clinical significance of this binding specificity was assessed in a 2-year clinical follow-up. DESIGN: Consecutive serum samples positive for antisperm antibodies by both enzyme-linked immunosorbent assay and immunobead testing were evaluated for acrosin-binding specificity. SETTING: The patients were followed in an outpatient setting by private infertility specialists. PATIENTS: Sixty-five consecutive infertile referral patients with positive antisperm antibody were evaluated. Clinical follow-up was obtained on 8 of 9 females with evidence of antibody binding to acrosin and 19 of 26 females with no specific binding to acrosin. INTERVENTIONS: Prednisone therapy was given during six courses of intrauterine insemination with husband's sperm. All treatment decisions were made by private physicians independent of the acrosin-binding result. MAIN OUTCOME MEASURES: Pregnancy status was obtained as part of a 2-year follow-up. RESULTS: Acrosin-binding specificity was demonstrated in 10 (15%) of the 65 patients. Two of the 8 women (25%) with antibody binding to acrosin and 6 of the 19 women (32%) with antisperm antibodies but no specific binding to acrosin delivered normal children. CONCLUSIONS: Although antibody-binding specificity to acrosin could be demonstrated, a 2-year clinical follow-up showed no difference in pregnancy rates when compared with women with antisperm antibodies showing no binding specificity to acrosin.     

Comparison of the tray agglutination technique with the gelatin agglutination technique for the detection of spermagglutinating activity in human sera.

Extensive comparisons were made between the tray agglutination test (TAT) and the gelatin agglutination test (GAT) in order to evaluate the validity of the TAT for demonstrating spermagglutinating activity in human sera. The incidence of spermagglutination in both tests was about the same. In screening 118 samples from infertile men and women, the TAT seemed to be more sensitive, whereas the reverse was true when 143 samples earlier found positive with the GAT were tested. These differences were mainly encountered with low titers. Two T-agglutinating sera (or 2% of the sera) with a GAT titer of 16 or more (both had a titer of 64) were missed with the TAT. On the other hand, two sera (or 2% of the screened samples) having small H-agglutinates up to a titer of 32 to 64 were positive only with the TAT.     

Chlamydia trachomatis infection in patients with laparoscopically verified acute salpingitis: Results of isolation and antibody determinations

Culture and serology studies have shown Chlamydia trachomatis (CT) to be one of the causes of acute salpingitis (AS). In the present investigation, results of cervical cultures were correlated with serum antibody titers to CT in patients with laparoscopically verified AS. Serum samples from 206 patients, including paired sera from 80, were assayed. Of 206 patients, 118 had chlamydial lgG antibody titers of 1:64 or more. Patients with negative cultures for CT and an lgG titer of 1:64 or more had a significantly higher geometric mean titer than corresponding patients with positive cultures. In paired sera, a seroconversion or a fourfold or greater rise in lgG titer to CT was demonstrated in 35%, while a further 11% had detectable lgM antibody in a titer of 1:8 or more. The overall isolation frequency of CT was 33%, compared with 19% for Neisseria gonorrhoeae.     

Application of staphylococcal protein A for immunodiagnosis of early toxoplasma infection]

RH strain Toxoplasma gondii was used to inoculate mice and rats. Sera collected at intervals were assayed by the indirect fluorescent antibody test (IFAT) and the latex agglutination test (LAT) to monitor the time-course change in titers of anti-toxoplasma IgM and IgG antibodies. In addition, the above sera and some anti-toxoplasma seropositive human sera (with IFAT antibody titers greater than or equal to 1:16) were adsorbed with staphylococcal protein A (SpA) and assayed for changes in IgM and IgG antibody titers so as to evaluate the usefulness of SpA adsorption in detecting anti-toxoplasma IgM which shows up early in toxoplasma infections. Samples assayed included 262 sera from mice, 65 sera from 5 rats and 85 human sera. The results revealed that parasite specific IgM-IFAT antibodies were detectable in 40% of the sera from mice 3 days after infection. After SpA adsorption, however, the IgM-IFAT antibody could be detected in a few specimens as early as 2 days post infection. IgG-IFAT and LAT antibodies first appeared in the sera on the 5th day of infection. In mice inoculated with freeze-killed tachyzoites (immunized mice), IgG-IFAT and LAT antibodies remained high from 16-35 days after inoculation, whereas IgM-IFAT antibodies were undetectable. Even after SpA treatment, only about half of the 16-day samples showed IgM antibody titers and the other specimens still remained negative. In general, IgM antibody titers increased 2-6 fold after SpA adsorption while IgG antibodies were almost completely removed after the treatment with a residual IgG-IFAT titer of less than or equal to 1:4.(ABSTRACT TRUNCATED AT 250 WORDS)     

Immune abnormalities in endometriosis compromising fertility in IVF-ET patients.

OBJECTIVE: To evaluate the incidence of immune abnormalities in patients with endometriosis and primary or secondary infertility. STUDY DESIGN: This study analyzed the incidence of alloantibodies and autoantibodies in 100 women with endometriosis and 62 patients with unexplained infertility without endometriosis who enrolled in an assisted reproduction program at the Colombian Fertility and Sterility Center from January 1, 1996, to May 30, 1997. The alloimmune status of the women was determined by testing for the presence or absence of antileukocyte antibodies. The autoimmune studies included antinuclear antibodies, antiphospholipid antibodies and lupus anticoagulant antibody. RESULTS: Negative titers of IgG antipaternal antibodies were identified in 34% of patients with primary infertility and in 34% of women with secondary infertility and a history of pregnancy losses. Positive titers of antinuclear antibodies were found in 27% (27/100) of the group of patients with endometriosis; of them, 30% (15/50) had primary infertility and 24% (12/50), secondary infertility. The average titer was 1/80. Forty-eight percent of the infertile patients (48/100) showed titers of antiphospholipid antibodies for IgG and IgM; 46% of these patients had primary infertility (23/50) and 50% (25/50), secondary infertility. This was significantly higher than in controls (P < .05). Two patients were positive for lupus anticoagulant antibody. In the group of patients with unexplained infertility without endometriosis, the incidence of antinuclear antibodies was 17.7% and of antiphospholipid antibodies, 30.6%. CONCLUSION: For women with endometriosis, alloimmune and autoimmune evaluation is recommended prior to their undergoing assisted reproduction in order to provide appropriate therapy for each case.     

Serum ABO immune antibodies in 1944 pregnant women

The ABO+ Rh blood groups and serum anti- A and anti -B anti bodies were examined in 1944 pregnant women. The results showed that 475 cases (24.4%) were positive for serum immune antibodies resulting in ABO incompatibility, and in 68 (3.4%) newborns haemolytic disease (HDN) occurred. The incidence of ABO blood group incompatibility increased with the increase of maternal serum immune antibody titer, the difference between different titers was significant (chi 2 = 4.13, P less than 0.05). Pregnant women with a high titer of serum immune antibody greater than 1:64 should be followed up regularly and those greater than 1:128 should be treated immediately. The relationship between positive serum immune antibody and blood group was discussed. We consider it is important to do maternal serum immune antibodies examination for antenatal care.     

Effect of therapy on infertile couples with antisperm antibodies

One hundred seventy-eight couples with positive antisperm antibody titers in serum and genital secretions were offered treatment with prednisone. Of 60 couples who received prednisone only, 43% conceived. Of 25 who had no therapy, 48% conceived. Fifty-four patients treated with prednisone received additional therapy and 31% conceived. Ten of 39 patients not treated with prednisone but receiving other therapies conceived. Cytotoxic antibodies were reduced in 30% to 42% of serum samples and in 24% to 33% of genital secretion samples. In those couples with decreased cytotoxic antibodies pregnancy rates were 40% to 60% compared with 0% to 23% in those with decreased hemagglutinating antibody titers. Our data suggest that prednisone did not improve overall pregnancy rates; pregnancy rates were comparable in both groups treated with other therapies; donor insemination was the most successful of the alternative therapies; reduction of cytotoxic antibody titers after prednisone treatment was associated with increased pregnancy rates.     

不孕症患者的自身免疫抗体分析

目的：探讨不孕症与患者自身免疫抗体的相关性。方法：采用酶联免疫吸附实验（ELISA）检测516例不孕症患者的血清抗精子抗体（ASAb）、抗卵巢抗体（AOAb）、抗子宫内膜抗体（EMAb）、抗绒毛膜促性腺激素抗体（hCGAb）及抗心磷脂抗体（ACAb）。选择125例正常妊娠妇女作为对照组．比较不孕症组与对照组5项抗体的阳性率。结果：不孕症组ASAb,AOAb,EMAb,hCGAb和ACAb的阳性率分别为26．7％,24．8％,25．2％,21．7％和21．3％。明显高于正常对照组4．8％,2．4％,1．6％,3．2％和1．6％（P均〈0．001）;5种抗体联合检测阳性率为38．2％,与分别单独检测这5项抗体的阳性率相比,差异均有统计学意义（P均〈0．05）。结论：ASAb,AOAb,EMAb,hCGAb和ACAb与不孕症有密切相关性,是引起不孕的重要原因。检测这5项抗体可作为不孕症病因诊断的一项指标。     

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。     

精浆内抗精子抗体和免疫复合物的研究——对男性不育患者及输精管粘堵术者的观察

本文用ELISA法研究了174名男性不育患者及43名输精管粘堵术后精浆中的抗精子抗体及其种类。结果表明,精浆中抗精子抗体的检出率,在男性不育患者中为24.1%,输精管粘堵术后者为48.8%。在这些患者的精浆中可检出IgA、IgG和IgM型抗精子抗体,其中主要是IgA。同组人群血清和精浆中的抗精子抗体缺乏相关性。精浆中抗精子抗体的存在与精子活动率的下降有一定的关系。应用PEG法测定精浆中免疫复合物,在66例精子抗体阳性的患者中,阳性率为56.1%,表明局部抗精子的自身免疫反应,在一定条件下可导致免疫复合物的形成。