M.-L. Windt

Treatment of male sperm autoimmunity by using the gamete intrafallopian transfer procedure with washed spermatozoa

Sixteen couples were diagnosed as having immunological infertility. To detect sperm-bound immunoglobulin (Ig), i.e., IgA, IgG, and IgM antibodies, the direct immunobead test (IBT) was used. In each individual patient, the direct IBT was greater than or equal to 70% positive for either IgA or IgG or both. The indirect IBT was positive for IgA and IgG antibodies in the serum of all the patients. Semen was collected in 15 mL medium (Hams F10 [Gibco, Grand Island, NY] + 10% whole blood serum) and prepared with the wash and swim-up method. Patients in the study group were treated for their immunological infertility problem by performing the gamete intrafallopian transfer (GIFT) procedure. An ongoing pregnancy was achieved in 7 of the 16 (43%) couples treated with the GIFT procedure with an ongoing pregnancy rate of 24.1% (7 of 29) per cycle. The GIFT procedure appears to be an effective and safe way of treating male immunological infertility.

EFFECT OF SPERM WASHING AND SWIM-UP ON ANTIBODIES BOUND TO SPERM MEMBRANE: USE OF IMMUNOBEAD/SPERM CERVICAL MUCUS CONTACT TESTS

The direct immunobead test (IBT) and the sperm cervical mucus contact (SCMC) test were used to evaluate the effect of sperm washes and swim-up on antibodies bound to the sperm membrane in 11 patients with autosperm antibodies (30-100% IgA and 30-100% IgG) but otherwise normal semen measurements. The tests were performed on semen samples before and after a wash/swim-up procedure in Ham F10 + 10% human blood serum. Sperm-bound antibodies in washed spermatozoa that were able to swim into a layer of medium did not differ from those in unwashed samples. The IBT and the SCMC test on unwashed and washed spermatozoa were also not significantly different. However, all other washed semen parameters were improved. Correct washing of semen samples is important when performing the IBT to prevent false negative results. Eight of the 11 partners (73%) of the patients tested became pregnant after treatment with washed sperm. The ongoing pregnancy rate was 64%.

Fertilization and pregnancy using metaphase I oocytes in an intracytoplasmic sperm injection program

AbstractPurpose: In this study we investigated whether metaphase I oocytes collected in an intracytoplasmic sperm injection program could successfully be matured and fertilized by injecting aged (>20-hr) spermatozoa. Materials and Methods: Metaphase I oocytes aspirated were preincubated for 20 hr to allow the oocytes to reach meiotic maturity. Only metaphase II oocytes were injected. The original sperm sample processed on the day of aspiration was used in the microinjection process. Results: One hundred eighty-three oocytes were collected, of which 42 (23%) were metaphase I oocytes. These were incubated for 20 hr and microinjected with the original sperm sample. Thirty-one (74%) of the metaphase I oocytes reached meiotic maturity (extruded polar body); 67.7% showed two pronuclei 18 hr after injection and 61.3% embryo development 40 hr postinjection. No difference in fertilization and embryo development rate was found in metaphase II oocytes injected 6 hr postaspiration versus 20 hr postaspiration. An ongoing pregnancy was also achieved using only embryos obtained from matured metaphase I oocytes. Conclusions: Metaphase I oocytes can be successfully maturedin vitro and injected using aged (>20-hr) sperm samples. Matured metaphase I oocytes, if successfully injected, produce embryos able to induce pregnancy.

Detection of sperm antibodies on unwashed spermatozoa with the immunobead test : a comparison of results with the routine method and seminal plasma TAT titers and SCMC test

ABSTRACT: This study investigated whether the immunobead test (IBT) could, for the purposes of simplicity and saving time, be applied directly on an unwashed semen sample instead of washed spermatozoa. These two methods were performed simultaneously on the semen samples of 15 men with a positive MAR test and 10 men with a negative MAR test. A possible association was found between the unwashed samples, showing positive IB binding (>20%) on the tail and/or head and the seminal plasma TAT titers (P < .00001, Fishers exact test). In all cases with IB binding of ≥20% on unwashed spermatozoa, positive seminal plasma TAT titers (≥32) and SCMC tests (≥50%) were found.

Effect of rapid dilution of semen on sperm-bound autoantibodies

The effect of rapid dilution of autoantibody-positive semen on sperm-bound antibodies was studied in 12 male patients (0-100% IgA and 10-100% IgG). The direct immunobead test (IBT) and the sperm cervical mucus contact (SCMC) test were used to detect sperm-bound antibodies on spermatozoa before and after rapid dilution and swim up in HAM F-10 + 10% human blood serum. All patients tested had normal semen parameters. Sperm-bound antibodies detected after swim-up of semen samples ejaculated into 15 ml HAM F-10 + 10% serum (diluted) did not differ significantly from undiluted samples. Most values were lower (IBT and SCMC) but were not statistically significant. Sperm motility, forward progression, and morphology were statistically improved. Pregnancies resulting from washed semen (diluted) in combination with artificial insemination (AIH) or gamete intrafallopian tube transfer (GIFT) took place in spite of antibodies still present on the sperm membrane and can probably be attributed to improved semen quality, minimizing of ovum-sperm distance, and cervical mucus elimination.

Evaluation of immunobead test (IBT), tray-agglutination test (TAT), and sperm immobilization test (SIT).

The present investigation relates the results obtained by the indirect immunobead test (IBT) to those found during the tray-agglutination test (TAT) and sperm immobilization test (SIT) performed on 16 positive blood sera samples. An IB binding of greater than 50% can be regarded as significantly positive (p less than 0.05), since 77% of the positive IB samples with an IB binding of greater than 50% also revealed a TAT titer of greater than 1:8. The IBT is an excellent test for the detection of sperm antibodies during routine screening procedures.

Antisperm Antibody Tests: Traditional Methods Compared to Elisa

Enzyme-linked immunosorbent assays (ELISAs) to detect sperm antibodies can be a useful addition to other tests. ELISAs allow more quantitative and detailed information than some other tests, but results should be compared to those of other tests. This study was conducted to correlate results of an ELISA that used a sperm membrane extract as antigen with those of other tests. Semen, seminal plasma, and serum of 34 sperm-antibody-positive and 36 sperm-antibody-negative men were tested. There was poor correlation between the ELISA results and those of the other tests. Only the results of a direct immunobead test done on semen correlated fairly well with results from ELISAs done on serum. Further studies are needed on fertility-specific antigens that can be identified and isolated and then studied by means of ELISAs.

Use of specific monoclonal antibodies to secretory IgA for the detection of spermatozoal antibodies in serum and seminal plasma by enzyme-linked immunosorbent assay

ABSTRACT: The role of anti‐sperm secretory IgA has recently received attention since some workers feel it plays an important role in the prognosis of the immunologically infertile couple. Current methods used in our laboratory cannot separately detect anti‐sperm secretory IgA. An enzyme‐linked immunosorbent assay (ELISA) utilizing specific monoclonal antibodies to secretory IgA was used to detect anti‐sperm secretory IgA as well as anti‐sperm monomeric IgA and IgG in serum and seminal plasma of a sperm‐antibody‐positive (ASA+) and sperm‐antibody‐negative (ASA‐) group of men. Results showed significantly raised serum levels in the ASA+ group when compared to the ASA‐group for anti‐sperm secretory IgA (P < .001), anti‐sperm monomeric IgA (P < .001), and anti‐sperm IgG (P < .01). Seminal plasma levels were also raised in the ASA + group, but only significantly so for monomeric IgA (P < .02). The performed ELISA has definite potential in research, especially with the use of monoclonal antibodies for the detection of anti‐sperm secretory IgA, but cannot as yet be used as a prognostic predictor of fertility in the individual antibody‐positive patient. Infertility specific antigens will have to be identified and isolated and subsequently used in the ELISA.