Antisperm antibodies and in vitro fertilization

The purpose of this study was to investigate the influence of antisperm antibodies in the male, the female, or both partners on the outcome of in vitro fertilization treatment. The results in terms of ongoing pregnancies in the male and female antibody-positive group were the same as in the antibody-negative group. In the double antibody-positive group two of the three patients became pregnant. When high levels of antisperm antibodies were present on the spermatozoa, the fertilization rate was significantly reduced. In the female positive group no clear relationship between the antibody titer and the fertilization percentage could be detected. Abnormal semen quality was responsible for a much lower fertilization rate than the presence of antibodies. The conclusion of this study is that in vitro fertilization provides an equal chance of conception in couples with antisperm antibodies in comparison with couples with no antibodies if the other semen parameters are normal.     

Normal fertilization in men with high antibody sperm binding by the addition of sufficient unbound sperm in vitro

A high incidence of fertilization failure has been reported in men with over 70% of their sperm bound with isoantibodies. In three men with greater than 80% antisperm antibody binding with IgG and IgA immunolobulins, a normal rate of fertilization (29/46 oocytes; 60%) was achieved by adding a sufficient number of motile sperm to provide at least 50,000 unbound sperm per oocyte. This method appears to be simple and more effective than attempting to separate unbound sperm in vitro.     

Relation between antisperm antibodies and the rate of fertilization of human oocytes in vitro

To clarify further the role of antisperm antibodies in in vitro fertilization, the occurrence of antisperm antibodies on ejaculated sperm and in sera was determined by the immunobead binding assay in 67 couples after an unsuccessful in vitro fertilization cycle. Antisperm antibodies in maternal sera were associated with a failure of oocyte fertilization (P <0.02) or with fertilization of only 9–19% of the oocytes (P <0.01) in vitro. Antisperm antibodies were detected in sera from 13 of 24 women (54.2%) where no fertilization occurred, 9 of 14 women (64.3%) where less than 20% of the oocytes fertilized, and 3 of 19 women (15.8%) where greater than 40% of the oocytes fertilized. Antisperm antibodies in these sera were mostly IgG and directed against the sperm tail. Antibodies on the surface of ejaculated motile sperm were also associated with a low (9–19%) fertilization rate (P <0.01). Sperm-bound antibodies were detected in 2 of 24 men (8.3%) where no fertilization occurred, 5 of 14 men (35.7%) where less than 20% of the oocytes fertilized, and 0 of 19 men where fertilization was greater than 40%. Sperm-bound antibodies were mainly IgA and were tail-directed. Antisperm antibodies in sera of males were not related to the rate of fertilization. Antisperm antibodies were detected in female partners of 21 of 46 couples (45.7%) with unexplained infertility, 2 of 12 women (16.7%) with blocked tubes, 4 of 7 women (57.1%) with endometriosis, and 0 of 2 women with adrenal hyperplasia. There was no relation between the fertilization rate and the maternal age, number of oocytes harvested, or semen quality. We conclude that antisperm antibodies are present in sera from a high percentage of women with unexplained infertility and that antibodies reacting with sperm tails may directly interfere with fertilization in vitro or may be a surrogate marker for another factor that interferes with this event.     

In vitro DNA fluorescence after in vitro fertilization (IVF) failure

Purpose A pilot study was performed to test the diagnostic value of in vitro DNA fluorescence in oocytes that failed to fertilize after IVF. Ten patients with a cleavage rate less than 20% after IVF were included.Results Uncleaved oocytes were observed by fluorescence microscopy after incubation with the DNA fluorescent dye Hoechst 33342. Four main causes which may have contributed to the low cleavage rate were found: (1) sperm incapacity to penetrate the oocyte despite the absence of the usual criteria for male infertility, (2) oocyte immaturity, (3) delayed fertilization, and (4) oocyte abnormalities revealed by aberrations in the morphology of the female chromatin.Conclusions The possibility of a rapid and detailed analysis of the maturational status of unfertilized oocytes, the morphology of the female chromatin, the presence and quantity of spermatozoa tightly bound to the zona pellucida, and sperm penetration into the oocyte without subsequent pronucleus formation, using DNA fluorescence, allows us to clarify further the cause of fertilization failure and to orient infertility treatment toward the male, the female, or both partners.     

Generation of mouse oocyte monoclonal isoantibodies: their effects and those of antisperm monoclonal antibodies on in vitro fertilization

Monoclonal isoantibodies to mouse oocyte antigens were generated by modified hybridoma techniques similar to those described for mouse sperm monoclonals. Following isoimmunization with mouse oocytes and cell fusion, hybrid cells were cultured initially in a semi-solid medium containing methylcellulose. Seven to ten days after cell fusion about 350 hybrid clones were recovered for subculture. By an indirect immunofluorescence assay using frozen or fresh mouse oocytes, twenty hybridomas were shown to produce antibodies that bind to various oocyte components including antigens of the zona pellucida. However, they did not cross-react with mouse spermatozoa or lymphocytes.A system was established to evaluate whether monoclonal antibodies to gamete-specific antigens have any inhibitory effects on the fertilization of mouse oocytes in vitro. A monoclonal antibody against zona antigen(s), ME 56, was shown to block fertilization of mouse oocytes via the inhibition of sperm binding to the zona pellucida. On the other hand, three out of four antibodies reacting with mouse sperm acrosomes were also inhibitory to mouse in vitro fertilization, perhaps mainly due to the inhibition of sperm acrosomal reactions. Using a sodium dodecylsulfate gel/protein blot radioimmunobinding method, the molecular weight of zona antigen(s) that react with ME 56 was determined to be in the range of 95,000, whereas that of the acrosomal antigen(s) reacting with the fertilization-inhibiting antibody, MS 207, was about 30,000. The results of this preliminary study suggest that monoclonal antibodies to certain gamete antigens can be a valuable tool for the analysis of sperm-egg interactions during the fertilization processes.     

Results of in vitro fertilization in women with antisperm antibodies in serum,cervical mucus,and follicular fluid

This study was undertaken to investigate the presence of antisperm antibodies (ASA) in serum, cervical mucus, and follicular fluid (FF) of women undergoing in vitro fertilization and embryo transfer (IVF-ET). IgG and IgA ASA directed mostly against sperm head were found at similar concentrations in serum, cervical mucus, and FF of 2 of 34 patients. Ninety-one percent fertilization and 100% cleavage rates, respectively, were observed in one of the two patients. No fertilization occurred in the second patient. In both women, in vitro sperm penetration tests revealed hostile mucus and repeated postcoital tests were poor. It is concluded that the sperm-cervical mucus penetration test and mucus ASA measurements are useful in establishing the diagnosis of immunological infertility.     

Failure of fertilization in in vitro fertilization: The “Occult” male factor

Failure of fertilization in patients undergoing in vitro fertilization (IVF) deserves extensive analysis for better prediction of the success or failure of this therapeutic modality. Consequently, we retrospectively studied the 52 couples in whom fertilization failed during Norfolk series 18 to 25, in an effort to establish the precise causes of failure. In the initial evaluation, pure oocyte abnormalities were identified in 19.2% of the cases; 32.6% showed sperm abnormalities, and a combination of oocyte and sperm anomalies was found in 7.7%. In 40.4% of the cases, failure of fertilization could not be explained. Re-assessment of sperm morphology by new, strict criteria increased the identification of sperm abnormalities to 61.5% and of combined sperm and oocyte anomalies to 13.4%, for a total of 74.9% of sperm factors involved, as opposed to 40.3% in the original evaluation. The incidence of unexplained failed fertilization was substantially reduced, to 11.5%. In a control group (tubal infertility) matched by age, date, and stimulation, in whom fertilization occurred, 83.3% had normal sperm parameters as judged by the new criteria for morphology evaluation. This paper emphasizes the need for a more accurate diagnosis of sperm abnormalities to establish the true incidence of this factor in failed fertilization and to obtain information of prognostic value to patients and clinicians.     

Intracytoplasmic sperm injection overcomes previous fertilization failure with conventional in vitro fertilization

Objective: To evaluate the outcome of intracytoplasmic sperm injection (ICSI) in patients with previous idiopathic fertilization failure (≤20% fertilization rate) after conventional IVF.

Design: Retrospective analysis.

Setting: IVF program at a university medical center.

Patient(s): Twenty-five patients who underwent 38 ICSI cycles after experiencing unexplained fertilization failure with conventional IVF (group A) and 87 patients who underwent 118 ICSI cycles for male factor indications during the same period (group B).

Intervention(s): Intracytoplasmic sperm injection was performed in a subsequent cycle after fertilization failure with conventional IVF.

Main Outcome Measure(s): Outcomes of IVF were compared between groups A and B.

Result(s): Fertilization was achieved with ICSI in all patients with previous fertilization failure. The mean (±SD) fertilization rate (68% ± 21% vs. 64% ± 22%), implantation rate per embryo (22.6% vs. 20%), and delivery rate per cycle (47.3% vs. 49.1%) did not differ significantly between groups A and B. Overall, 72% of patients with previous unexplained fertilization failure had a successful pregnancy after ICSI.

Conclusion(s): Intracytoplasmic sperm injection can overcome unexplained fertilization failure caused by a potentially occult gamete abnormality, with the same fertilization, implantation, and pregnancy rates as are seen in patients with abnormal sperm parameters.     

Antiphospholipid antibodies and pregnancy rates and outcome in in vitro fertilization patients

Objective: To determine the relationship between antiphospholipid antibodies and pregnancy rates (PRs) and outcome among IVF patients.

Design: Prospective collection of all serum samples with assays for immunoglobulin G (IgG), IgA, and IgM antibodies for anticardiolipin, antiphosphatidyl serine, antiphosphatidyl ethanolamine, antiphosphatidyl choline, antiphosphatidyl inositol, antiphosphatidyl glycerol, and anti-phosphatidic acid being done following completion of all treatment cycles.

Setting: A tertiary care teaching hospital.

Patient(s): Seven hundred ninety-three patients attempting to conceive through IVF.

Main Outcome Measure(s): Pregnancy rates (PRs) and pregnancy loss rates relative to each of the various antiphospholipid antibodies that were measured.

Result(s): There were 528 pregnancies for an overall PR of 66%. Pregnancy rates were equal among patients with positive and negative antiphospholipid antibodies for each of the 21 measured antibodies. Use of receiver operator characteristic curves and logistic regression further confirmed that there was no relationship between PRs or outcome based on antiphospholipid antibodies for any definable threshold value.

Conclusion(s): Elevated antiphospholipid antibody levels are not associated with any change in PRs or pregnancy loss rates in patients attempting to conceive through IVF.     

Serum unconjugated bisphenol A concentrations in women may adversely influence oocyte quality during in vitro fertilization

Bisphenol A (BPA) is an endocrine disruptor with estrogenic properties that can adversely affect meiotic spindle assemblies. Our data indicate that BPA exposure in female patients may interfere with oocyte quality during IVF, as suggested by the inverse association between serum unconjugated BPA concentration and normal fertilization.     

Ultrasonic detection of a cumulus oophorus in patients undergoing in vitro fertilization

Ultrasonic studies for the detection of a cumulus oophorus were carried out in 57 women taking part in an in vitro fertilization (IVF) program. When intrafollicular echoes were dissociated, clearly prominent from the follicular wall, they were considered to be a sure cumulus mass, and when they were only slightly prominent, they were suspected to be a nondissociated cumulus. All patients had at least one ultrasonically visible cumulus. A cumulus was seen in 50% of the follicles and 70% of them were dissociated. Cumuluses were also seen in follicles <18 mm in diameter but a significantly higher number of them were not clearly dissociated. The number of observed dissociated cumuluses correlated significantly with the number of recovered mature oocytes. However, in 18 patients there were more mature oocytes retrieved than cumuluses identified by ultrasound. When a cumulus mass is seen, it can be taken as evidence of a sign of maturity of that particular follicle and oocyte. However, mature oocytes are also found where no cumulus was seen by ultrasound. Lack of visible cumulus has little significance in predicting the maturity of the oocyte.     

Cytotoxic sperm antibodies and in vitro fertilization of mature oocytes: A preliminary report

The fertilization rates of mature oocytes during in vitro fertilization and embryo transfer (IVF-ET) using fetal cord serum-supplemented insemination media were 57% for five infertile couples without sperm antibodies (group 1). But they were 50% for four of nine infertile couples (group 2) with cytotoxic sperm antibodies in both partners (n=6) or the husband alone (n=3). Two women in group 1 were successful in achieving normal, full-term pregnancies with the delivery of normal infants (²=4.2, P < 0.05, by chi-square analysis). One of them consistently tested negative for sperm antibodies, while her husband was previously treated with antibiotics for infection and transient sperm antibodies in the seminal plasma. Subsequently, antibody liters in the husband were in the normal range when the successful IVF-ET was performed. One woman in group 2, with antibodies to her autoimmune husband's sperm but not control sperm and with a long-standing poor postcoital test sperm motility, conceived through artificial insemination with donor sperm (AID) after failing to conceive with her husband through IVF-ET. These data suggest that the presence of cytotoxic sperm antibodies in the serum and/ or secretions of both partners reduces the rates of fertilization of mature oocytes in spite of using fetal cord serum in the IVF media. Pregnancy achievement is impaired in this group.     

Intracytoplasmic sperm injection as a treatment for unexplained total fertilization failure or low fertilization after conventional in vitro fertilization

OBJECTIVE: To determine whether IVF or intracytoplasmic sperm injection (ICSI) should be the choice of treatment in case of a previous IVF attempt with unexplained total fertilization failure or low fertilization (<25%). DESIGN: Prospective study. SETTING: Leiden University Medical Center. PATIENT(S): Thirty-eight couples undergoing IVF and ICSI on sibling oocytes after a first IVF attempt with total fertilization failure or with low fertilization (<25%). INTERVENTION(S): Performing IVF and ICSI on sibling oocytes. MAIN OUTCOME MEASURE(S): Fertilization and (ongoing) pregnancy rate. RESULT(S): A total of 271 oocytes were collected in 24 oocyte retrievals in the total fertilization failure group. Hundred nine oocytes were randomly allocated to IVF and 12 were fertilized (11%); 162 sibling oocytes were allocated to ICSI and 78 were fertilized (48%). In 8 of the 24 patients fertilization occurred after IVF. The pregnancy rate after transfer of 1 IVF and 1 ICSI embryo (n = 3) was 67% and after the transfer of 2 ICSI embryos (n = 21) this was 52%. In the low fertilization group 169 oocytes were collected in 14 oocyte retrievals. Seventy-two oocytes were randomly allocated to IVF and 16 were fertilized (22%). Ninety-seven sibling oocytes were allocated to ICSI and 58 were fertilized (60%). In 7 of 14 patients fertilization occurred after IVF. The pregnancy rate after the transfer of 1 IVF and 1 ICSI embryo (n = 5) was 80% and after the transfer of 2 ICSI embryos (n = 9) this was 33%. CONCLUSION(S): Performing ICSI on some oocytes of a cohort may avoid total fertilization failures both in patients with a history of total fertilization failure and in patients with a history of low fertilization, as the percentage of fertilization is higher after ICSI compared to IVF and the recurrence of total fertilization failure and low fertilization is high after IVF treatment.     

Artificial oocyte activation improves cycles with prospects of ICSI fertilization failure: a sibling oocyte control study

Research questionDoes artificial oocyte activation improve clinical outcomes for patients at risk of intracytoplasmic sperm injection (ICSI) fertilization failure?DesignIn this study, sibling oocytes from patients with previous ICSI failure or severe teratozoospermia were divided equally into two groups, half for artificial oocyte activation (AOA) with ionomycin after conventional ICSI and the other half for conventional ICSI only (non-AOA). The fertilization rates, cleavage rates, transferable embryo rates and blastulation rates of the two groups were compared first; the clinical pregnancy and live birth rates were also compared to assess the efficiency and safety of AOA.ResultThe outcomes of the AOA group were significantly better than those of the conventional ICSI group in terms of the fertilization (50.38% versus 33.86%, respectively, P < 0.001), cleavage (59.16% versus 39.04%, respectively, P < 0.001) and transferable embryo rates (43.51% versus 26.69%, respectively, P < 0.001). The blastulation (43.53% versus 36.11%, respectively), implantation (26.83% versus 15.79%, respectively), clinical pregnancy (38.46% versus 25%, respectively) and live birth rates (38.46% versus 16.67%, respectively) were not significantly different.ConclusionThis study showed that AOA improved some aspects of cycles at risk of ICSI failure by increasing the fertilization and transferable embryo rates. But blastulation, pregnancy and implantation rates were not improved. The study is limited by its small size and absence of data on cumulative outcomes.     

Blocking of human fertilization in vitro by sera with sperm-immobilizing antibodies

Serum samples with sperm-immobilizing antibody activity from six women were examined for ability to block sperm-egg interaction by a zona penetration test where human follicular ova matured in vitro were used. Exposure of spermatozoa from a fertile healthy donor to the sera impaired binding to and penetration through the zona pellucida of the spermatozoa completely in five cases and incompletely in one case. Successful fertilization in vitro was achieved by using fetal cord serum instead of autoserum of the patient included in the in vitro fertilization and embryo transfer program. These results suggest that interference with sperm-egg interaction may be an additional mechanism of infertility that is caused by antisperm antibodies.     

An ultrastructural analysis of an oocyte from an in vitro fertilization patient with repeated polyspermic fertilization

Objective: Our goal was to determine any ultrastructural anomalies in an oocyte from a patient with a history of polyspermy. Results: Ultrastructural observations of the cortical ooplasm of several oocytes from each of three control patients showed a large population of intact cortical granules. Conversely, one oocyte from a patient with repeated polyspermic fertilization contained a relative paucity of granules in the cortex. Quantitative analysis of the cortices of control oocytes indicated that there were 17.02±0.52 cortical granules present per measured field of view, compared with 4.40±2.92 granules per field in the other oocyte. Conclusions: The presence of sufficient cortical granules is necessary for normal (monospermic) fertilization to occur. When contrasted to the cortical granule population of oocytes from several control patients, the cortex of one oocyte from the other patient showed few of these organelles. Therefore, the absence of a sufficient number of granules may have precluded normal fertilization from occurring in the eggs of this patient.     

The association between chlamydial-specific IgG and IgA antibodies and pregnancy outcome in an in vitro fertilization program

Chammydial-specfic IgG and IgA antibodies were determined by a single serovar (L2) immunoperoxidase assay (IPA) in the serum of all patients that have conceived in an in vitro fertilization and embryo transfer (IVF & ET) progrom (n=106) and in a group of patients that went through the program at the same period of time and did not conceive (n=94). The prevalence rate of elevated IPA IgG (titers1128) and IPA IgA (titers116) specific to chlamydiae was significantly higher (P<0.001) in the IVF&ET pregnancy loss and nonconception groups (failures) versus the IVF&ET term pregnancy group (successes) (74 vs 47%, odds ratio=4.1, and 34 vs 14%, odds ratio=4.3, respectively). Stepwise discriminant analysis revealed that elevated specific chlamydial IgG had the greatest effect on the variance between successes and failures in this study group. Our study indicates the possible role of past or chronic active chlamydiae infection on the take-home baby rate in an IVF&ET program.     

Effect of Chlamydial antibodies on the outcome of in vitro fertilization (IVF) treatment

Objective Our objective was to study the relationship between the presence of Chlamydia trachomatisantibodies and the success of IVF treatment.Design We evaluated 118 in vitro fertilization and embryo transfer (IVF-ET) treatment cycles from 51 couples with a history of infertility lasting for at least 2 years. All women starting a treatment cycle had their serum chlamydial antibody titers measured by indirect immunofluorescent technique. All patients received similar ovarian stimulation regimens and the oocytes collected were inseminated with similar concentrations of motile sperm. Clinical data from couples where the female partner had C. trachomatisab titers 40 have been compared with the equivalent data from couples where the female partner had C. trachomatisab titers <40.Results There was no statistically significant difference between the two groups concerning age, infertility period, oocytes collected, oocytes fertilized, and fertilization rate, and the pregnancy rates were comparable.Conclusion Previous exposure to C. trachomatisdid not alter the success rate of IVF-ET.