Sperm Cervical Mucus Contact Test Research Articles

The casual effect of lifestyle factors on outcomes of assisted reproductive techniques: a protocol study on Iranian infertile couples

BackgroundAttention to additional factors affecting the success of assisted reproduction techniques (ARTs) is very important and with appropriate interventions in some of these factors, success rate can be as high as improve a lot. There is a lot of evidence that lifestyle factors can influence on ARTs outcomes. Current knowledge of the effect of combined effect of several lifestyle factors on the outcomes of ARTs is low and contradictory. The importance of the causality of this phenomenon is felt by the pandemic of inappropriate lifestyle as well as the dramatic increase in infertility in the world. The aim of this cohort study is to scrutinize the casual effect of a specific range of contemporary lifestyle factors on ARTs outcomes.MethodsA prospective cohort study will be conducted in Royan institute, Tehran, Iran. Each infertile couple will be monitored from the first visit to the end of treatment cycle. The data will be collected electronically and include the following: detailed interview about lifestyle and socioeconomic status, past medical history, general physical examination, assessment of menstrual and ovulatory status, assessment of tuboperitoneal, assessment of uterine, assessment of cervix, urogenital examination, semen analysis, anti-sperm antibodies, biochemical analysis, sperm-cervical mucus contact test, in vitro cervical mucus penetration test and sperm functional assays. To estimate the casual effect of lifestyle variables on clinical pregnancy and live birth, the obtained propensity score (PS) from generalized boosted models (GBM) will be matched between couple with and without live birth.DiscussionThis is the first study to prospectively obtain detailed information on causes of ARTs success. Determining the casual effect of lifestyle variables on ARTs success will be important to inform strategies most likely to increase the success rates in ARTs.

The SCMC test: a screening test for spermatozoal antibodies.

The sperm cervical mucus contact test (SCMC-test) were performed on 62 couples on account of their post coital test results. Fifteen of the 17 couples (88%) who repeatedly demonstrated unexpected poor post coital tests, revealed a positive SCMC-test. In 10 cases the SCMC test results were confirmed by the tray-agglutination and mixed antiglobulin reaction. The SCMC-test is considered to be a reliable screening test for the detection of spermantibodies amongst infertile couples.

Citation classics in Fertility and Sterility, 1975–2004

Citation classics in Fertility and Sterility, 1975–2004

Analysis of spermatozoa from the proximal vas deferens of vasectomized men.

This study assessed the condition of spermatozoa from the proximal vas deferens of men after vasectomy. The fluids of both proximal vas deferens were collected from 67 vasectomized men by cannulating the vas deferens at the time of vasectomy reversal. Selected sperm parameters were analysed after incubation of the spermatozoa for 30 min at 37 degrees C. Sperm concentration in the proximal vas from vasectomized men (16,312 +/- 21,496 million per ml, geometric mean: 7948 +/- 398 million per ml) was significantly higher than that of fertile men and was maintained at a constant level independent of the duration of vas obstruction. The means of sperm motility (36.2 +/- 26.2%), spermatozoa with normal morphology (50.7 +/- 21.7%), sperm viability (53.0 +/- 25.3%) and hypo-osmotic swelling test (HOS-test, 53.9 +/- 21.7%) were statistically lower than the respective values for normal fertile men. There was no significant correlation between the duration of vas obstruction and the above semen parameters. In 46.4% of vas fluids all spermatozoa were immotile and this condition was more common after 3 years of vasectomy. Immotile spermatozoa in the proximal vas fluids at the time of vasectomy reversal may be an important factor for predicting semen quality and fertilizing ability after vasovasostomy. There were no significant differences in the results of sperm-cervical mucus penetration test (CMPT) between spermatozoa from vasectomized and fertile men. Antisperm antibodies on the surface of spermatozoa from the vas of vasectomized men were determined by the immunobead test (IBT; 78.6% for IgG, 32.1% for IgA) and sperm cervical mucus contact test (SCMC, 36.4%).(ABSTRACT TRUNCATED AT 250 WORDS)

Analysis of spermatozoa from the proximal vas deferens of fertile men.

Fluids from the left and right proximal vas deferens were collected from 105 normal fertile men by cannulating the vas deferens during vasectomy, and sperm parameters analysed. Sperm motility (73.1 +/- 13.3%), normal sperm morphology (75.2 +/- 11.1%), sperm viability (72.7 +/- 18.8%) and the hypo-osmotic swelling test (73.3 +/- 19.2%) were in the normal range, compared with that of ejaculated spermatozoa. However, sperm concentration in the proximal vas deferens (6274.6 +/- 5103.8 x 10(6) ml-1 was higher than that in semen. Sperm concentration in the right vas deferens was significantly higher (P < 0.05) than that in the left and the percentage of spermatozoa showing abnormal cervical mucus penetration was significantly higher (47%) for the left than for the right (18%). There were no anti-sperm antibodies on the surface of spermatozoa from the vas deferens as determined by the sperm cervical mucus contact test and immunobead test. These parameters of spermatozoa from the proximal vas may reflect those of spermatozoa from the human cauda epididymis.

The significance of antisperm antibodies for sperm-cervical mucus interaction.

An overview is presented of the effects of antisperm antibodies on the sperm-cervical mucus interaction. Antisperm IgA on spermatozoa or in cervical mucus can severely inhibit sperm penetration of cervical mucus and migration through it. Disturbance of the sperm-cervical mucus interaction is the only firmly established effect of these antisperm antibodies and leads to reduced fertility, as shown by a poor or negative result of the post-coital test. The presence of antisperm IgA in the male or female partner can be investigated more specifically with the sperm-cervical mucus contact test and can be confirmed by the sperm agglutination test on bromelinliquefied cervical mucus, by the mixed antiglobulin reaction test or by the immunobead test for IgA.

Chlamydial infection—a female and/or male infertility factor?

After screening a large series (n = 491) of asymptomatic males of infertile partnerships for chlamydial immunoglobulin (Ig) G antibodies (Chlam AB), no significant influence of past chlamydial infection was found with regard to semen analysis, postcoital testing, in vitro sperm-cervical mucus penetration tests with hormonally standardized cervical mucus, circulating antisperm antibodies (detected with three different methods), local IgG and IgA antibodies (detected by means of the mixed antiglobulin reaction test) on the sperm surface, the sperm-cervical mucus contact test, and a microbial screening of semen samples for mycoplasmas and other potentially pathogenic micro-organisms. However, when the findings were correlated with infertility factors of patients' female partners and the subsequent pregnancy rate in a prospective study, a significant positive correlation of male Chlam AB with a tubal factor in their wives as cause of the couple's infertility was found. The results suggest that the main influence of Chlamydia trachomatis on male fertility is based on sexual transmission and negative influence on tubal function of female partners, but not on reduced sperm functional capacity.

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%.

Sperm cervical mucus contact test and immunobead test for sperm antibodies.

Twenty-five patients were studied for the presence of sperm antibodies in their cervical mucus and seminal plasma. The sperm cervical mucus contact test (SCMC) and the immunobead test (IBT) were used to evaluate sperm antibody activity. A close correlation was found between the results of the SCMC and IBT tests, and it was concluded that both techniques can be regarded as good indicators of sperm antibodies.

The Significance of the Fc Part of Antispermatozoal Antibodies for the Shaking Phenomenon in the Sperm-Cervical Mucus Contact Test

Donor spermatozoa with good motility were pretreated with four sera containing high titers of sperm-agglutinating IgG, one serum without sperm-agglutinating activity, the IgG fractions from these five sera, F(ab)2 and Fragment antigen binding (Fab) fragments from these sera and from the IgG fractions, and one seminal plasma sample with a high titer of sperm-agglutinating IgA. With mixed antiglobulin reaction tests the percentage of motile pretreated spermatozoa sensitized with IgG Fab or IgG Fragment crystalline (Fc) parts was determined. Spermatozoa sensitized with intact antispermatozoal IgG showed a strong reduction in their capacity to penetrate cervical mucus. The reduction of the penetration capacity was determined by estimation of the percentage of motile spermatozoa rapidly shaking (S%) in the sperm-cervical mucus contact (SCMC) test. Removal of the Fc parts resulted in a decreased S%. Treatment of spermatozoa, on which Fab fragments were present, with intact antibodies to IgG Fab fragments, resulted in a recurrence of a high S%. A decrease of the S% was also found if Fab fragments from antibodies to IgG Fc fragments were added to spermatozoa sensitized with intact antispermatozoal IgG. Similarly, it was found that a decrease of the S% occurred when IgA sensitized spermatozoa were treated with Fab fragments from antibodies to human IgA. In the sperm penetration meter test the IgA sensitized spermatozoa treated with Fab fragments from anti-human IgA antibodies showed a better penetration than untreated IgA-sensitized spermatozoa.

Induction of the shaking phenomenon by pretreatment of spermatozoa with sera containing antispermatozoal antibodies

Sera containing sperm-agglutinating and/or complement-dependent sperm-immobilizing activity were tested for their ability to induce a shaking phenomenon in the sperm-cervical mucus contact (SCMC) test. Donor spermatozoa were treated with the sera in a one-step incubation and washing procedure. The percentage of motile pretreated spermatozoa showing the shaking phenomenon (S%) was determined in the SCMC test. In addition, the immunoglobulin class of antibodies present on the pretreated spermatozoa was determined in mixed antiglobulin reaction tests for IgG, IgM, and IgA. An S% of at least 80 was always observed with spermatozoa pretreated in sera with a sperm agglutination titer of at least 32, provided that IgG was detected on more than 90% of the motile spermatozoa. The high S% was also obtained with spermatozoa pretreated in purified IgG from sera containing antispermatozoal antibody activity. It was concluded that the reduced cervical mucus penetration capacity of spermatozoa pretreated with sera containing antispermatozoal IgG could be ascribed, at least partially, to the occurrence of a shaking phenomenon.

5 - Immunology of Infertility

Recent research on immunological infertility in men and women is reviewed and the possibilities for therapeutic success in this area are assessed. Surface antigens of the acrosome and main tail piece appear to provoke antibodies of special relevance to male and female infertility and are recognized by circulating sperm-immobilizing antibodies in women and by immobilizing and agglutinizing antibodies in men. Assessment methods have focused on the development of tests of local immunity to sperm. Antisperm antibodies have been tested via sperm microagglutination, the gelatin agglutination test, the sperm immobilization test, and immunofluorescence techniques. In addition, measurement has focused on antibodies in cervical mucus, antibodies in seminal plasma, and cell-mediated immunity. Methods involving both partners include postcoital test, the sperm-cervical mucus penetration test, and the sperm-cervical mucus contact test. There remains a need for the development of specific radioimmunoassys for the precise detection and quantitation of antibodies to sperm antigens, especially those of cell membrane origin. In males, autoimmunity to sperm antigens can be related to infertility by 2 main pathogenic mechanisms: 1) the adverse effects of antibodies directly on spermatozoa, and 2) the association with disordered spermatogenesis resulting in oligospermia and azoospermia. In women, the effector pathways of local immunization mediate both systemic and cell-mediated immune responses. Local antibodies can interfere with the reproductive process by arming macrophages and enhancing phagocytic clearance of spermatozoa from the genital tract, mediating cytotoxic effects on sperm, preventing sperm from adequately penetrating cervical mucus, intefering with sperm capacitation, and influencing sperm selection within the female genital tract. Between 5-10% of infertile men and women show evidence of anitbodies to sperm. Treatment has included occlusion therapy, intrauterine insemination, immunosuppression, and spermatogenesis suppression. There is a need for prospective controlled assessment of the therapeuatic approaches currently in use, as well as for research on the role of cell-mediated immunity.

Immunoglobulin class of antispermatozoal antibodies from infertile men and inhibition of in vitro sperm penetration into cervical mucus.

The presence of IgG and IgA on motile spermatozoa from normal semen donors and men from infertile couples was studied with mixed antiglobulin reaction (MAR) tests. The percentage of motile spermatozoa with IgG (IgG MAR %) was found to be related to the circulating antispermatozoal IgG. No direct relation could be detected between the IgG MAR % and the sperm agglutinating activity in seminal plasma (SP) or the percentage of motile spermatozoa showing the shaking phenomenon (S%) in the sperm cervical mucus contact (SCMC) test. The percentage of motile spermatozoa with IgA (IgA MAR %) showed no direct relation to the sperm agglutinating activity in serum and SP, but was roughly proportional with the S%. It was discerned that the shaking phenomenon in the SCMC test was probably due to presence of IgA on the motile spermatozoa. Previously it had been demonstrated that also the sperm agglutinating activity in SP is caused by IgA that is probably locally produced in the male genital tract. In conclusion it was thought that the reduced ability of penetration into cervical mucus by spermatozoa from infertile men is caused by locally produced IgA.

Spermagglutinating Antibodies and Sperm Penetration of Cervical Mucus from Infertile Women with Spermagglutinating Antibodies in Serum

Spermagglutinins were demonstrated by the tray agglutination technique in cervical mucus collected during presumably-ovulatory cycles in 8 women among 21 patients with spermagglutinating antibodies in serum treated for infertility. A "poor" sperm penetration test was recorded exclusively in women with spermagglutinins in cervical mucus, and the results of the sperm-cervical mucus contact test were significantly correlated to the spermagglutinin titers in cervical mucus. The incidence of spermagglutinating antibodies in cervical mucus from infertile women was estimated to be 2.2% on the basis of the results in the present study. However, an inhibiting effect on sperm penetration in cervical mucus by spermagglutinins is expected to occur in less than 1% of women from infertile couples. A decrease in spermagglutinin titers in cervical mucus observed during estrogen medication was significantly associated with improved sperm penetration in vitro. The latter results may indicate a new approach to the treatment of infertility due to the presence of spermagglutinating antibodies in cervical mucus.

Functional Role of Spermagglutinating Antibodies in Men

The presence of spermagglutinating antibodies in blood and semen has been associated with reduced fertility in men. In this study 203 men were screened for infertility. An antibody measurement was performed on the blood and semen of each patient. Each semen specimen was also examined for spontaneous agglutination. Twelve patients with a positive reaction in the blood underwent a sperm-cervical mucus contact test. We have attempted to correlate the findings in blood with the observed functional results in semen and cervical mucus. We believe that these simple tests should be included in the infertility work-up in order to determine the presence of immune infertility.

Presence of Sperm Agglutinating Antibodies in Infertile Men and Inhibition of in vitro Sperm Penetration into Cervical Mucus

The relation between presence of antispermatozoal antibodies in infertile men and the inhibition of the in vitro sperm penetration into cervical mucus (CM) was studied with the sperm cervical mucus contact (SCMC) test. The tests were performed with semen from infertile men and from semen donors. The CM used permitted good penetration of normal spermatozoa. The so called “shaking phenomenon”, the result of a specific interaction of spermatozoa and CM, was expressed in the shaking percentage (S%).The S% did not change beyond the experimental error within 30 min after mixing semen and CM. The S% was 30 at the most in 194 out of 198 SCMC tests with normal donor semen and normal pre‐ovulatory CM. Significant negative correlations (P &lt; 0.005) were found between the readings of the sperm penetration meter (SPM) test on one hand and the S%, the sperm agglutination titer in the serum and the sperm agglutination titer in the seminal plasma (SP) on the other hand. Significant positive correlations (P &lt; 0.005) were found between the S% and the sperm agglutination titer in the SP. The sperm agglutination titer in serum and in SP correlated significantly better (P &lt; 0.02) with the S% in the SCMC test than with the readings of the SPM test.It was concluded that: 1. A high S% is highly specific for the presence of antispermatozoal antibodies in infertile men, 2. The SCMC test is more suited than the SPM test for studying the effect of antispermatozoal antibodies on the penetration and migration of spermatozoa into CM,

Immunology

Immunology

The Sperm-Cervical Mucus Contact Test: A Preliminary Report

Infertility in men and women with spermagglutinins is the result of disturbed penetration and migration of spermatozoa in the cervical mucus. In ejaculates with partial spermagglutination caused by autoimmunization, the progressive propulsion of the sperm was changed into stationary, shaking movement the moment the sperm came into contact with cervical mucus. The same alteration in spermatozoal motility pattern also occurred when spermatozoa from a normal, fertile ejaculate came into contact with cervical mucus of a woman whose serum contained sperm antibodies. This shaking phenomenon was visualized in a simple test, the sperm-cervical mucus contact test. We demonstrated that sensitized spermatozoa exhibit the shaking phenomenon after contact with the glycoprotein fraction of the cervical mucus and not after contact with the aqueous fraction. Therefore, the hypothesis is introduced that the shaking phenomenon is due to an interaction between sensitized spermatozoa and the glycoprotein micelles in cervical mucus.