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PubMed1976

[Sperm agglutination, sperm immobilization and sperm immunofluorescence test in married couples with unexplained infertility (author's transl)].

Thirty-four married couples with clinically unexplained infertility and normal spermiograms in all male partners were investigated for sperm antibodies by means of the sperm agglutination, sperm immobilization and sperm immunofluorescence test. In 50% of the couples, sperm antibodies were found in at least one of the partners and with at least one of the tests, e.g. ten times in the female partner alone, six times in the male partner alone, and once in both partners. The correspondance of the three tests was astonishingly low; it is assumed, therefore, that they indicate different types of antibodies. On the basis of the present investigations it is not possible to assess which type of antibody is responsible for the infertility of the female or male partner respectively. This seems only possible when the decrease of certain antibody types can be determined after a period of no antigen contact (avoidance of sperm contact) and when conception has taken place. The significance of autoantibodies against sperm in clinically healthy men with normal spermiograms remains open.

Antibodies↗
PubMed1979

Isolation of human spermatozoa membrane antigens binding sperm-immobilizing and sperm-agglutinating antibodies.

Peptides of human spermatozoa were dissolved with Hyamine 2389 and Triton X-100 and separated by chromatography on Biogel P4 columns into seven fractions. The antigenic activities of the separated sperm-membrane fractions were tested according to their capacity to inhibit sperm agglutination and sperm immobilization (immune inhibition test) in human sera of sterile patients. Sperm-agglutinating and sperm-immobilizing activity was tested by the microtray agglutination and microtray immobilization test. A titer reduction was achieved only in sperm-immobilizing sera. Four sperm antigenic fractions revealed in the majority of the repeatedly tested sperm-immobilizing sera an inhibition of the antigen-antibody reaction. No reaction was observed after exhaustive absorption of the tested seven antigen fractions with sperm-agglutinating sera. Therefore the conclusion can be drawn that sperm-agglutinating and sperm-immobilizing antibodies react with different sperm antigens. Normal human sera without sperm antibodies served as control. As no sperm agglutination or sperm immobilization was obtained after absorption of these control sera our antigen fractions do not produce sperm agglutination or sperm immobilization.

Antigens, Surface↗
PubMed1980

Sperm immunofluorescence in infertile men: Its relationship with sperm agglutination and gel agglutination.

The spermatozoa from 10 infertile male patients all with sperm agglutination were subjected to direct immunofluorescence using antisera raised against IgA, IgM, and IgG. Sera from these patients were subjected to the Gel Agglutination Test. Although the Gel Agglutination Test related closely with sperm agglutination, none of the spermatozoa from these patients produced a repeatable positive response to direct immunofluorescence. This study suggests that neither sperm agglutination nor the Gel Agglutination Test demonstrate the presence of sperm specific antibodies in the semen of these infertile men.

Agglutination Tests↗
PubMed1990

[Sperm agglutination antibodies studied in infertile couples at the Women's Clinic in Plzen 1980-1988].

The authors assessed sperm-agglutination antibodies in infertile couples in 1980-1988, using the Kibrick test: in sera from 699 female patients, a positive result was found in 142 (20%) women, in sera from 352 men, in 6 (1.7%) sperm-agglutination antibodies were detected. Using the microagglutination test, they examined sera of 3501 sterile women; in 273 (6.8%) they found sperm agglutination up to 1:16, above titre 1:32 in 12 (0.4%) of the patients. Using the same test, they examined sera of 2803 men and found sperm-agglutination up to 1:16 in 21 (0.7%) and above 1:32 in 6 (0.3%). The most frequent type of sperm agglutination is aggregation of the heads; aggregation by tails or mixed agglutination are less frequent. In 3252 sterile women where no sperm-agglutination antibodies were found Kremer's test was negative in 1145 patients. The authors analyzed in 196 sterile women the ovulation mucus by the microagglutination test and found a high anti-spermatozoidal activity in 24 (12%) women. Concurrently an indirect MAR test was made. Antibodies against sperm cells were found in 24 (12.2%) of specimens in sIgA, in 8 (4.1%) in IgG, in 2 (2%) in IgM and in one sample (0.5%) in IgE. The indirect MAR-test in sera of 325 sterile women revealed a predominance of anti-spermatozoidal antibodies in IgG, in men in IgA. In seminal plasma the authors detected antibodies against sperm cells in sIgA. In seminal plasma the authors detected antibodies against sperm cells in sIgA and IgE.

Antibodies↗
PubMed1995

Relationship between the direct mixed antiglobulin reaction (MAR) test and spontaneous sperm agglutination in men from infertile couples.

To determine the relationship between the direct MAR test and spontaneous sperm agglutination in men from infertile couples, semen samples from 160 men were studied. Spontaneous sperm agglutination was present in 80 samples, and absent in the others. Routine semen analysis and the direct MAR test were performed on all samples. The incidence of positive MAR tests in samples with and without spontaneous sperm agglutination were 15 and 0 per cent respectively (p < 0.05). There were no significant differences in mean sperm concentration, motility, and morphology between the samples with and without spontaneous sperm agglutination, and between the positive and negative MAR test groups. It can be concluded that spontaneous sperm agglutination relates significantly to antisperm antibodies detected by the direct MAR test, and this factor could be used as indication for antisperm antibody testing of infertile men.

Coombs Test↗
PubMed1975

Difficulty of obtaining reproducibility in the Franklin and Dukes test for the detection of sperm-agglutinating antibodies in human sera.

The Franklin and Dukes (FD) test is used as a screening test for the determination of sperm-agglutinating antibodies and as a proof of therapeutic effects in cases of infertility, but various parameters of the FD test remain unclear and the application of different techniques has revealed highly different results. To test the statistical evidence for the FD technique, conditions regarded as optimal prerequisites were applied: a sperm concentration of 20 times 10(6)/ml was produced with Baker's buffer and serum dilutions were made with Baker's buffer, starting at 1:4. Only the standard deviation of parallel tests with one serum sample and one semen sample on the same day was found to be within an acceptable range. Using ejaculates of the same donor and the same serum sample on different days gave results that were not reproducible. The FD test should not, therefore, be used for quantification of sperm-agglutination antibodies except for a comparison in one test with one semen sample on the same day. Although the FD test only allowed a qualitative evaluation of sperm-agglutinating antibodies the % sperm agglutination was more informative than agglutination titres. For quantification of sperm-agglutinating antibodies, the FD test should be replaced by other techniques.

Agglutination Tests↗
PubMedDate not supplied

Monoclonal antibody inducing human sperm agglutination.

A mouse fused splenic cell line producing antibody that induces agglutination of human sperm was cultivated. The antibody was localized to the midpiece and tail of human spermatozoa by an indirect immunofluorescence method. The interacting antigen was identified in a sperm membrane protein preparation by SDS gel/protein blot immunofluorescence method. The molecular weight of the antigen was in the vicinity of 84 Kdaltons.

Animals↗
PubMedDate not supplied

Characterization of sperm agglutinating monoclonal antibody and purification of the human sperm antigen.

A monoclonal antibody capable of inducing tail-to-tail agglutination of human sperm was purified by ammonium sulfate fractionation and chromatography on DEAE-cellulose. A 75- to 100-fold purification was achieved. The antibody was of the IgG class and composed of gamma 1 heavy chain and lambda light chains. The purified preparation was effective in inducing sperm agglutination at a minimum concentration of 0.55 microgram/mL and was active after storage at 4 degrees C for over 3 months in the presence of sodium azide. The sperm antigen interacting with this YWK-1 antibody was purified by immunoaffinity chromatography on Sepharose 4B conjugated with YWK-1 antibodies. Two interacting proteins were isolated, with estimated Mr of 84 and 20 kD. The 84 kD protein had an isoelectric point in the range of pH 5.7 to 6.1, and probably lacks carbohydrate moieties. The 20 and 84 kD components may share common antigenic determinants.

Antibodies, Monoclonal↗
PubMed1980

The significance of serum sperm-agglutinating antibodies in men with infertile marriages.

Three hundred consecutive male partners of infertile marriages had serum samples tested for sperm agglutinating and sperm immobilising antibodies, using a micro-technique: 30.3% were found to be positive. Out of a control group of 100 fertile men 10 had antibodies detected (10%) and these at low titre; this difference was significant. The results of these serum tests were found to correlate with the results of seminal plasma tests, the mixed antiglobulin reaction for IgG and sperm cervical mucus contact testing, although all of these tests were less discriminatory with regard to fertility. The serum micro-immobilisation test showed a statistical association with fertility when analysed in those couples where both husband's and wife's tests showed no other bar to fertility.

Antibodies↗
PubMed1981

Postcoital tests and sperm-agglutinating antibodies in men.

Postcoital tests (PCTs) were examined from couples where the husbands had circulating head-to-head (H-H) or tail-to-tail (T-T) sperm-agglutinating antibodies. The results were compared with findings in couples without antibodies. The presence of H-H sperm-agglutinating antibodies did not interfere with the outcome of the PCT. High serum titers of T-T sperm-agglutinating antibodies, the presence of such antibodies in the seminal fluid, and/or a strong or complete autoagglutination in the husband's ejaculate markedly reduced the number of invading spermatozoa as well as their survival in the PCT. A moderate titer of T-T sperm-agglutinating antibodies was also reflected in the PCT but low titers did not have any influence on the results.

Autoantibodies↗
PubMed1986

A comparison of sperm agglutination and immobilization assays with a quantitative ELISA for anti-sperm antibody in serum.

An enzyme-linked immunosorbent assay (ELISA) that quantitates antisperm antibody in serum was compared with standard sperm agglutination and immobilization assays with the use of sera from 40 normal and 292 subfertile individuals. Quantitation of the assay was accomplished by standardizing assay parameters, including the incorporation of a standard reference curve, the number of whole target sperm, the optimal dilution of serum, the selection of microtiter plate, and the time and temperatures involved in the adsorption and incubation phases. With this method, the level of antisperm antibody binding to target sperm in 40 normal fertile individuals was found to be 2.3 (+/- 1.1 standard deviation [SD]) fg immunoglobulin (Ig)/sperm. An increased mean level of 7.4 +/- 3.7 fg Ig/sperm was determined in 84 infertile patients with positive agglutination and/or immobilization tests. In 208 individuals with negative agglutination and immobilization tests the mean concentration of antisperm antibody was 2.5 +/- 1.3 fg Ig/sperm. Postvasectomy patients assayed by this method had a mean Ig binding value of 7.1 +/- 2.4 fg Ig/sperm. The infertile group with positive agglutination and/or immobilization tests had a significantly higher mean antisperm antibody level than the normal fertile group, according to the Student's t-test for independent samples (P less than 0.001). This indirect serum-based assay reproducibly quantitates antisperm antibody binding to whole target sperm, suggests the normal and abnormal levels of antisperm antibody, and correlates with standard functional assays.

Antibodies↗
PubMed1987

A fertile man with a high sperm agglutination titer in the seminal plasma: a case report.

A fertile man had sperm-agglutinating activity in his serum (titers 1:16-1:128) and in his seminal plasma (titers 1:128-1:2048). The antibodies in the seminal plasma could be absorbed with anti-IgA antiserum but not with anti-IgG antiserum. A fresh ejaculate showed strong auto-agglutination of the spermatozoa. With mixed antiglobulin reaction tests (MART) and/or immunobead tests (IBT), IgA and IgG were detected on almost all motile spermatozoa; the erythrocytes, in the MART, and the latex spheres, in the IBT, adhered mainly to the tip of the tail. After mixing the fresh semen with cervical mucus, only 40% of the spermatozoa were locally shaking. The spermatozoa showed excellent penetration of cervical mucus in vitro. This case shows that IgA coating of the tails of the spermatozoa does not necessarily lead to adherence of these spermatozoa to the micelles of the cervical mucus and that the sperm cervical mucus contact test has a better predictive value than the sperm agglutination titer in the seminal plasma.

Adult↗
PubMed1984

Association between infertility following reversal of vasectomy and the presence of sperm agglutinating activity in semen.

A statistically significant association between infertility, following reversal of vasectomy, and the presence of sperm agglutinating activity in post-operative semen samples has been demonstrated in a retrospective study. Sperm agglutinating activity was measured using a microtitration assay, and the number of positive samples rose from 3/78 (4%) pre-operatively, to 34/134 (25%) post-operatively. High titres (greater than or equal to 1:32) of agglutinating activity were observed only when viable sperm were present in the ejaculate. Positive semen titres were obtained only when parallel serum samples were positive at titres of greater than or equal to 1:256. Conception only occurred in the partner of one man whose seminal plasma contained agglutinating activity, but 22/34 (65%) of the men whose semen lacked such activity were fertile.

Female↗
PubMed1978

Immune sperm agglutination: are only motile spermatozoa involved?

Human sperm-agglutinating sera was examined using the tray agglutination technique (TAT) with whole diluted semen, and also with a suspension of only progressively motile spermatozoa-obtained from the same semen sample by the penetration of motile spermatozoa in an overlying medium layer. The titres of tail-to-tail (T) agglutinating sera were considerably lower with the latter sperm suspension, but head-to-head (H) agglutinating sera gave similar titres with the two types of sperm suspensions. Attempts to increase titres of T-agglutinating sera with only progressively motile spermatozoa were made by reducing the progression rate of the spermatozoa, and by increasing the concentration of the sperm cells. However, significant titre differences remained. Moreover, the agglutinates obtained were very small, and difficult to observe with the routinely employed magnification of x40. The lower titres still remained where seminal plasma was used as the penetration medium, indicating that a loss of coating antigen(s) was not the cause of lower titres. It seems likely that spermatozoa are attached to each other only weakly in T agglutination, much more weakly than in H agglutination. It is considered that immotile sperm cells that are present in whole diluted semen reinforce the T agglutination process, producing the higher titres.

Antibodies↗