Antisperm Antibody Titers Research Articles

EBV transformation and microfusion as the potential source of human monoclonal antisperm antibodies.

Peripheral blood lymphocytes were obtained from vasectomized men with high serum titers of antisperm antibodies. An Epstein-Barr virus (EBV) transformation was performed either with B cells or mononuclear leukocytes. The effect of feeder cells (irradiated umbilical cord blood lymphocytes), cyclosporin A, and in vitro stimulation of lymphocytes with sperm extract on EBV transformation was evaluated. Antibody-producing cells were screened for specificity against human sperm by an enzyme-linked immunosorption assay (ELISA) one to six weeks after transformation. Using B cells or leukocyte mononuclear cells, we found that the percentage of wells containing antibody reactive against human sperm was greatest two weeks after transformation (range 3% to 7.5% positive wells). To increase and maintain antibody synthesis by these transformed cells, microfusions were performed in those wells positive for antisperm antibody using the UC 729-6 lymphoblastoid cell partner. Then resultant hybridomas were expanded, subcloned, and preliminarily characterized.

Clinical significance of unilateral testicular obstruction in subfertile males.

Unilateral testicular obstruction has been defined by exploratory scrototomy in 80 spontaneously infertile males. The most common sites of obstruction were either in the tail of the epididymis after previous infection (40 cases) or in the vas deferens due to congenital absence or after previous infection or surgery (24 cases). Half of the patients had severe oligozoospermia (less than 5 million/ml) and three-quarters had developed antisperm antibodies, often in high titres. Spermatogenesis was shown by testicular biopsy to be normal (Johnsen score-count greater than 8.0) in almost all of the obstructed testes, and serum FSH levels were normal. Following surgical reconstruction, with prednisolone therapy if indicated by high antisperm antibody titres, 19 (32%) of 60 patients with adequate follow-up successfully impregnated their female partners, with the best results occurring in 30 men who started with sperm counts less than 5 million/ml, 12 of whom (40%) were successful. Removal of irreparably blocked testes in 10 men led to profound falls in high antisperm antibody titres, with production of two pregnancies. Unilateral testicular obstruction appears to be a correctable cause of infertility in some human males.

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

To better understand the immunogenic 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.

A Study on Antisperm Antibody in Homosexual Men

Using a hemagglutination assay we surveyed antibody distribution to human sperm antigen in a sexually transmitted disease (STD) clinic population and in an AIDS study group. We found no significant difference for the mean antisperm antibody titres between sexually active heterosexual men, women or homosexual men, homosexual men with or without AIDS, and no correlation between T4/T8 lymphocyte ratios and antibody titres in homosexual men. We conclude that levels of antibody to sperm antigens are not significantly greater in homosexual men and antisperm antibody levels do not correlate with the diagnosis of AIDS.

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.

Effect of cyclosporin A in male autoimmune infertility

Nine infertile men (mean duration of infertility, 8.3 years) with antisperm antibodies were treated with the immunosuppressive drug cyclosporin A for 6 months with a dose of 5 to 10 mg/kg/day. Seminal plasma and serum antisperm antibodies fell in three subjects on treatment, and sperm count and motility increased substantially in one. Three successful pregnancies occurred in the study group: one on treatment, one in the first cycle of artificial insemination with the husband's sperm after treatment (twin infants), and one 3 months after cessation of treatment (twin infants). Successful conceptions with cyclosporin were unrelated to falls in antisperm antibody titer.

Why Does Vasectomy Reversal Fail?

The scrotum was explored in 32 patients who had vasectomy reversal which failed to restore their fertility. 23 patients were azoospermic and 9 had severe or persistent oligozoospermia. Among the 23 azoospermic cases the vasa were found to be blocked in 12; following repeat vaso-vasostomy normal sperm counts were produced in 10 patients; 8 of these patients followed up for 6 months or more successfully impregnated their wives. In 4 cases there was a secondary epididymal block; following epididymo-vasostomy 3 of 4 patients otained normal sperm counts and 2 wives became pregnat. In 7 patients there was a very high antisperm antibody titstet (<1024 by tray agglutination test) and despite reparative surgery and prednisolone therapy no pregnancies were obtained. The overall pregnancy rate for this group was 7 of 19 (37%). Anong 9 oligozoospermic cases unilateral obstruction was found in 8. The block was found in the vas in 5 and in the epididymis in 2 and in 2 there were very high antisperm antibody titers. After reconstructive surgery 4 patients had normal sperm counts and 1 pregnancy has been obtained. (authors)

Cyclic prednisolone therapy for male infertility associated with autoantibodies to spermatozoa

Seventy-six subfertile men with significant titers of antisperm antibodies were treated with a new corticosteroid regimen, consisting of prednisolone, 40 mg daily, rising to 80 mg daily if antibody titers did not fall, given from days 1 to 10 of the partner's menstrual cycle, for up to nine cycles. Twenty-five (33%) of the partners became pregnant during a treatment cycle, more than twice the expected incidence without treatment. No serious complications occurred, although one half of the patients had transient minor side effects. This regimen appears to be encouraging and suitable for further assessment in a prospective controlled trial.

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.

Mechanisms of injury in unilateral testis torsion.

We investigated the mechanisms of injury involved in unilateral testis torsion as well as the interventional effects of orchiectomy at 48 h and immunosuppression with corticosteroids in rats. Torsion was associated with abnormal contralateral testis histology and raised antilymphocytes and antisperm antibody titers. Both orchiectomy and steroid administration lessened these findings, suggesting that an immunological process underlies the abnormality seen in this experiment model.

Why does vasectomy reversal fail?

The scrotum was explored in 32 patients who had vasectomy reversal which failed to restore their fertility. Twenty-three patients were azoospermic and nine had severe or persistent oligozoospermia. Amongst the 23 azoospermic cases, the vasa were found to be blocked in 12; following repeat vaso-vasostomy, normal sperm counts were produced in 10 patients; 8 of these patients followed up for 6 months or more successfully impregnated their wives. In four cases there was a secondary epididymal block: following epididymo-vasostomy, three of four patients obtained normal sperm counts and two wives became pregnant. In seven patients there was a very high antisperm antibody titre (greater than 1024 by tray agglutination test) and despite reparative surgery and prednisolone therapy, no pregnancies were obtained. The overall pregnancy rate for this group was 7 of 19 (37%). Amongst nine oligozoospermic cases, unilateral obstruction was found in eight. The block was found in the vas in five and in the epididymis in two, and in two there were very high antisperm antibody titres. After reconstructive surgery, four patients had normal sperm counts and one pregnancy has been obtained.

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.

Long-term effect of vasectomy on coronary heart disease

We investigated the association between coronary heart disease (CHD) and vasectomy in a population of 10,632 men who were under surveillance for multiple CHD risk factors during participation in a university-based exercise testing program. We conducted a mail survey with telephone follow-up to determine the vasectomy status of individuals in the population. Responses were obtained from 6,159 individuals. The 4.944 males on whom information was complete enough to be included in the multivariate analysis comprised the study population. Among the 1,383 (28 per cent) vasectomized males in the study populations, the interval from vasectomy to the time of the survey ranged from less than one year to 37 years with a mean duration of 15 years. Although increased relative risks for CHD were found to be associated with family history of CHD, high blood pressure and smoking in this population, the relative risk of CHD associated with vasectomy was not increased in general, nor was it increased when the vasectomized males were classified by time since vasectomy. Likewise, serum antisperm-antibody titers were not predictive of CHD among vasectomized men. These studies support the findings from previous investigations of populations with shorter average post-vasectomy experience in which vasectomy has been shown to be unassociated with altered risk of CHD in humans.

Long-term effect of vasectomy on coronary heart disease.

We investigated the association between coronary heart disease (CHD) and vasectomy in a population of 10,632 men who were under surveillance for multiple CHD risk factors during participation in a university-based exercise testing program. We conducted a mail survey with telephone follow-up to determine the vasectomy status of individuals in the population. Responses were obtained from 6,159 individuals. The 4.944 males on whom information was complete enough to be included in the multivariate analysis comprised the study population. Among the 1,383 (28 per cent) vasectomized males in the study populations, the interval from vasectomy to the time of the survey ranged from less than one year to 37 years with a mean duration of 15 years. Although increased relative risks for CHD were found to be associated with family history of CHD, high blood pressure and smoking in this population, the relative risk of CHD associated with vasectomy was not increased in general, nor was it increased when the vasectomized males were classified by time since vasectomy. Likewise, serum antisperm-antibody titers were not predictive of CHD among vasectomized men. These studies support the findings from previous investigations of populations with shorter average post-vasectomy experience in which vasectomy has been shown to be unassociated with altered risk of CHD in humans.

Anti-sperm antibodies and fertility of turkey hens

Anti-sperm antibody titers increase with time in serum of turkey hens following a standard production schedule of artificial insemination (AI). In hens receiving intravenous (IV) or intraperitoneal (IP) additional AI, serum anti-sperm antibody levels increase more rapidly after a lag phase. A single injury to the oviduct also resulted in increased anti-sperm antibodies similar to IV and IP groups. This is a new observation that a single injury increased antibody titers to spermatozoa equal in IV and/or IP injections. A negative correlation between serum anti-sperm antibody titers for IV, IP and injury to oviduct and fertility of these groups was observed. Hens of IV and injury to oviduct groups with high levels of anti-sperm antibodies in the last 2 weeks of production had significantly lower fertility than hens with low levels of antibodies and control hens.

Immunity to Tumor-Associated Antigens in Vasectomized Men

Leukocytes from 49 vasectomized and 57 age-matched nonvasectomized men were tested in the leukocyte adherence inhibition (LAI) assay for reactivity to sperm and various 3-M KCl human tumor extracts. Forty-four percent of the vasectomized men and 15% of the control group were reactive to the sperm antigen preparation (P less than or equal to 0.03). Similarly, a significantly higher percentage of vasectomized men responded to 3 of 5 tumor extracts tested: melanoma I (34.7 vs. 15.8%, P less than or equal to 0.04), squamous cell carcinoma (48.8 vs. 26.0%, P less than or equal to 0.04), and breast carcinoma (19.5 vs. 4%, P less than or equal to 0.04). Thirty percent of vasectomized versus 4% of the control group responded to more than two tumor antigens (P less than or equal to 0.03). The degree of LAI reactivity to each tumor extract was highly correlated with degree of antisperm LAI reactivity, and the degree of LAI responsiveness to one of the melanoma extracts was significantly correlated with antisperm antibody titer as measured by the sperm-immobilization assay. Furthermore, nonresponsive leukocytes from the control population converted to tumor antigen-responsive when incubated with sera from vasectomized LaI-positive men. Data from this study indicated that a large percentage of vasectomized men with sperm immunity were responsive to tumor-associated antigens in the LAI test and that antisperm antibodies or other serum factors played a role in this response. These results and the evidence that most tumor antigen-responsive cancer patients also have serum antibodies that reacted with sperm suggested that vasectomized men and cancer patients frequently responded to immunologically cross-reactive antigenic determinants present on both sperm and tumor cells.

The incidence and influence of antisperm antibodies in infertile human couples on sperm-cervical mucus interactions and subsequent fertility

Analysis of serum samples from 698 infertile couples revealed antisperm antibodies present in 16.5% of the men and 21.6% of the women. Overall, 31.1% of the couples possessed at least one individual with positive results. Sperm-immobilized activity was detected in 29.6% of the cervical mucus (CM) samples from 459 women. Reduced sperm penetration of CM was significantly associated with serum titers of antisperm antibodies in both sexes and also with immobilizing activity in CM of women. The incidence of subsequent pregnancy in 376 infertile couples was reduced significantly if one or both partners had antisperm antibodies in serum or in genital tract secretions. The latter was reflected by evaluation of the immobilization, penetration, and shaking phenomenon of sperm in CM.

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Leukocytes from 49 vasectomized and 57 age-matched nonvasectomized men were tested in the leukocyte adherence inhibition (LAI) assay for reactivity to sperm and various 3-M KCl human tumor extracts. Forty-four percent of the vasectomized men and 15% of the control group were reactive to the sperm antigen preparation (P less than or equal to 0.03). Similarly, a significantly higher percentage of vasectomized men responded to 3 of 5 tumor extracts tested: melanoma I (34.7 vs. 15.8%, P less than or equal to 0.04), squamous cell carcinoma (48.8 vs. 26.0%, P less than or equal to 0.04), and breast carcinoma (19.5 vs. 4%, P less than or equal to 0.04). Thirty percent of vasectomized versus 4% of the control group responded to more than two tumor antigens (P less than or equal to 0.03). The degree of LAI reactivity to each tumor extract was highly correlated with degree of antisperm LAI reactivity, and the degree of LAI responsiveness to one of the melanoma extracts was significantly correlated with antisperm antibody titer as measured by the sperm-immobilization assay. Furthermore, nonresponsive leukocytes from the control population converted to tumor antigen-responsive when incubated with sera from vasectomized LaI-positive men. Data from this study indicated that a large percentage of vasectomized men with sperm immunity were responsive to tumor-associated antigens in the LAI test and that antisperm antibodies or other serum factors played a role in this response. These results and the evidence that most tumor antigen-responsive cancer patients also have serum antibodies that reacted with sperm suggested that vasectomized men and cancer patients frequently responded to immunologically cross-reactive antigenic determinants present on both sperm and tumor cells.

The frequency of serological anti-zona pellucida activity in males, females and children

945 human sera from children, fertile and infertile females, pregnant women and fertile and infertile males were tested for antibodies to porcine zona pellucida by immunofluorescence and the pronase-induced zona-lysis inhibition test. Positive sera were found in each of the groups examined, with a higher percentage of anti-zona activity in infertile, pregnant and menopausal women. There was no correlation between anti-sperm antibody titer and the incidence of anti-zona activity in either female or male human sera, nor did the occurrence of anti-zona activity correlate with a particular blood group type.

Immunoglobulin E levels and antisperm antibody titers in infertile couples

Immunoglobulin E (IgE) levels and antisperm antibody titers were determined in samples of serum and seminal plasma from 25 fertile men (Group A), 18 infertile men without measurable immunity to sperm (Group B), and 42 infertile men autoimmune to sperm (Group C), and in samples of serum and cervical and vaginal secretions from 25 fertile women (Group D), 28 infertile women without measurable immunity to sperm (Group E), and 32 infertile women isoimmune to sperm (Group F). Among the men, IgE levels in the serum, measured in international units per milliliter, were elevated in Group C (230 ± 41, mean ± SEM) as compared with Groups B (94 ± 33, P < 0.05) and A (55 ± 8, P < 0.001). In contrast, IgE levels in the seminal plasma in Group C (180 ± 44) were not significantly different from those in Group B (48 ± 21), but were higher than those in Group A (8 ± 2, P < 0.05). In the women, serum IgE levels were higher (P < 0.001) in Group F, with isoimmunity to sperm (472 ± 55), than in Groups E (219 ± 32) and D (76 ± 16). Wives of autoimmune husbands had somewhat, though not significantly, higher serum IgE levels (406 ± 55), than had wives of nonautoimmune husbands (247 ± 97). These results provide evidence for an elevated IgE response in subjects with significantly elevated antisperm antibody titers.