Tage Hjort

Antisperm antibodies and fertility after vasovasostomy: a follow-up study of 216 men.

A group of 216 vasovasostomized men were tested with the mixed antiglobulin reaction for immunoglobulin (Ig)G, IgA, and secretory IgA antisperm antibodies bound to the sperm membrane. Free antisperm antibodies in serum and seminal plasma were detected with the gelatin agglutination test and the tray agglutination test. The results were related to the conception rates. In a subgroup with a pure IgG response, the conception rate reached 85.7%, whereas only 42.9% of the men who also had IgA on the sperm induced pregnancy. When 100% of the spermatozoa were covered with IgA, the conception rate was reduced to 21.7%. The combination of IgA on all sperm and a strong immune response (titer in serum greater than or equal to 256) was associated with a conception rate of zero.

ASSOCIATION BETWEEN FAILURE TO IMPREGNATE AFTER VASOVASOSTOMY AND SPERM AGGLUTININS IN SEMEN

Sperm agglutinins emerging from the reopened part of the vas deferens were found in the seminal fluid of 10 out of 29 men after vasovasostomy. There was a strong association between the presence of these antibodies and the failure to impregnate over a period of 14 to 33 months. Of 20 couples, 11 pregnancies resulted among the 13 women whose men had no sperm agglutinins in the seminal fluid, but only 1 among the 7 whose men had such antibodies, 2p = 0.0044. The presence of sperm agglutinins in the seminal fluid was always associated with their presence in the serum, usually in a higher titre. Preoperative serum levels thus are highly predictive of ability to impregnate after reversal of vasectomy.

Anti-sperm antibodies, detected by agglutination, immobilization, microcytotoxicity and immunobead-binding assays

To determine the reliability of tests currently utilized in the detection of sperm-reactive antibodies, sera were provided as unknowns and studied without knowledge of the clinical histories. Four laboratories performed tray agglutination tests (TAT), three complement-dependent immobilization (SIT), and single laboratories sperm cytotoxicity (SCT), passive haemagglutination (PHA) and immunobead binding (IBB). Most investigators demonstrated an excellent correlation between duplicate sample results. Nearly all of the female sera were free of anti-sperm antibodies and positive results did not appear in greater frequency in women with unexplained infertility as compared with other categories. For the male sera, the highest incidence of anti-sperm antibodies in the infertile group (21% positive for sperm-reactive IgGs) was obtained by immunobead binding. The GAT and TAT results gave 7 and 12% positives, except for lower results in one laboratory. Sperm-reactive antibodies were detected most commonly in vasectomized men, with all assays except SCT and PHA. Of the newer techniques studied, IBB results correlated well with TAT, GAT and SIT, while SCT and PHA did not, suggesting that a different group of antibodies, perhaps directed against other sperm-associated antigens, was being detected by the latter procedures. In this light, emphasis was placed on the need to validate whether results of particular methodologies correlated with impaired sperm function and to develop methods that provided evidence for this premise, either on the basis of clinical criteria or altered gamete interaction in vitro.

Antibodies to spermatozoa and seminal plasma antigens detected by various enzyme-linked immunosorbent (ELISA) assays

Various ELISA methods have been applied by different research centers to test the efficiency of this approach for the diagnosis of sperm-immune infertility cases. The antigens used were either whole spermatozoa or solubilized spermatozoal membrane preparations and were immobilized on microtiter plates, except in one case where plastic beads were employed. Polyvalent second antibodies or protein-A labelled with enzymes served as tracers. A high frequency of positive sera was found in all groups including fertile controls with tests using whole spermatozoa as antigen. The methods using solubilized antigen preparations showed fewer positives on the whole and correlated better with the various clinical categories of the WHO sera. Whilst there was some agreement in the results between the various laboratories on a few sera, most of the positive sera found by one laboratory were reported as negative by others. More investigative work is needed to improve reproducibility between different laboratories and to reduce non-specific reactions with normal controls. A more precise definition of the proper cut-off levels for positives and negatives is also needed. Despite these short-comings, the development of an ELISA for the diagnosis of sperm-immune infertility cases seems to be justified in the long term.

Detection of autoimmunity to sperm: mixed antiglobulin reaction (MAR) test or sperm agglutination? A study on 537 men from infertile couples *

Two different ways of testing for antisperm antibodies were compared: the mixed antiglobulin reaction (MAR) test for demonstration of antibodies of the IgG and IgA classes bound in vivo to the sperm membrane antigens and the gelatin agglutination test for detection of nonbound antisperm antibodies in serum and seminal plasma. Samples from 537 men from infertile couples were investigated. Antibodies bound to the sperm membrane were detected in 49 men (9.1%), IgG in 44 (8.2%), and IgA in 38 cases (7.1%). Sperm agglutinins were recorded in seminal plasma from 30 men (5.6%) and in serum (titer greater than or equal to 16) from 43 men (8.0%). The investigation revealed a very close correlation between the results of MAR testing and the occurrence of sperm agglutinins in serum and seminal plasma. However, if one focuses on antisperm antibodies of the IgA class, which seem to play the major role in male immune infertility, the MAR test offered the advantage that a minor group of patients with pure IgG responses could be distinguished, and rare cases with mainly or exclusively locally produced IgA antibodies could be detected.

Spermagglutinating Antibodies and β-Spermagglutinins in Sera from Infertile and Fertile Women *

Sera from five groups of women were investigated by the tray agglutination technique for the presence of spermagglutinins: (1) 326 women from infertile couples, (2) 51 women in early pregnancy, (3) 65 women in advanced pregnancy, (4) 41 oral contraceptive users, and (5) 109 presumably fertile, nonpregnant women. By absorbing spermagglutinating sera with a cell-free eluate from spermatozoa, agglutination disappeared when due to a high-molecular weight compound with beta-mobility in preparative zone electrophoresis (denoted beta-spermagglutinin), whereas activity due to sperm antibodies was unchanged. Spermagglutinating sera occurred with comparable frequency among women from infertile couples, women in advanced pregnancy, and oral contraceptive users. The absorption study revealed agglutinating antibodies in a significantly higher frequency and in higher titers among women of infertile couples than among fertile women, in whom beta-spermagglutinins were found almost exclusively. These results demonstrate the importance of discriminating between sperm antibodies and beta-spermagglutinins in studies of sperm antibodies as a cause of infertility in women.

Autoantibodies against spermatozoal antigens detected by immunoblotting and agglutination. A longitudinal study of vasectomized males

Sera taken pre- and post-operatively at regular intervals within a year from 16 men undergoing vasectomy were analysed for autoantibodies against spermatozoal proteins by immunoblotting. The reaction patterns were compared with the results of sperm agglutination tests. Immunoblotting revealed the presence of autoantibodies against various spermatozoal polypeptides in all sera taken pre-operatively and post-operatively. On average, seven polypeptides showed reaction. During the post-operative period two patients developed spermatozoal agglutinins in moderate titers (greater than 16) but in immunoblotting no change in band reactivity was observed for these two patients. However, scanning of the immunoblotting results revealed that one of the patients, although without sperm agglutinins, during the post-operative period showed an increasing band colouring of a polypeptide of Mr 31,500, reflecting an increased level of the corresponding antibodies.

Antisperm antibodies on epididymal spermatozoa

ABSTRACT: Still under debate is the location in the genital tract where antisperm antibodies transuded from serum and locally produced antibodies, respectively, become attached to the surface of the spermatozoa. The mixed antiglobulin reaction (MAR) for immunoglobulin (Ig) G, IgA, and locally produced secretory IgA (sIgA) could be carried out on motile spermatozoa from epididymis from 13 of 34 men undergoing vasovasostomy. Four months and one year after the operation the MAR was repeated on a fresh semen sample from all patients. It was found that transuded as well as locally produced antisperm antibodies were bound to spermatozoa at the epididymal level of the genital tract. The follow‐up results on semen were almost identical to the results obtained with epididymal spermatozoa.

Evaluation of immunofluorescence on pig zona pellucida for detection of anti-zona antibodies in human sera

Using immunofluorescence tests, antibodies reacting with pig zone pellucida could be detected in 31% of 103 sera from patients in different clinically defined categories, mainly fertile males and females and patients from couples with unexplained infertility. The antibodies could in all cases be absorbed by means of pig erythrocytes - in most sera with all erythrocyte suspensions tested, but in some cases only with erythrocytes from certain animals - indicating that antibodies both to generally occurring pig antigens and alloantigens may be present in human sera. Staining of pig zonae was recorded equally frequently with unabsorbed sera from infertile and fertile males and females, respectively, but titres of 1:16 or more occurred more frequently among infertile than among fertile female (P = 0.053).

Do Autoantibodies to Sperm Reduce Fecundity? A Mini‐Review in Historical Perspective

PROBLEM: The paradox that early studies of antisperm antibodies in men showed a strong correlation between titers of circulating antibodies (essentially immunoglobulin [Ig] G) and reduction in conception rates, whereas more recent studies have indicated that the antifertility effect is mainly (or exclusively?) associated with IgA antibodies, impairing sperm migration through cervical mucus, was studied.