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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↗
PubMed1979

Sperm-agglutinating antibodies in infertile couples.

In a selected group of 150 infertile couples, sperm-agglutinating antibodies were examined by a microagglutination technique. Sperm-agglutinating activity was observed in sera from 12 men (8%) and 16 women (11%). Antibodies in seminal plasma were demonstrated in 8 men with correspondingly high serum titers. The type of agglutination in these cases was head to tail. Head-to-head agglutination was found in only 3 men, who all had agglutinating activity in serum only. An abnormal postcoital test was found in 57% of the couples, and 13% of the men and 14% of the women in these couples had agglutinating antibodies. A negative postcoital test was related more often to high than to low titer in female sera.

Agglutination Tests↗
PubMed1975

Sperm-agglutinating antibodies and testicular morphology in fifty-nine men with azoospermia or cryptozoospermia.

The relationship between the state of the germinal epithelium and the type and titer of circulating sperm-agglutinating antibodies has been investigated in a series of 59 azoospermic or occasionally cryptozoospermic men. The patients were grouped according to the condition of the germinal epithelium as observed from testicular biopsy specimens, as well as to type and titer of circulating sperm-agglutinating antibodies investigated by a previously described microagglutination technique. Evidence is presented to suggest that the presence of mature spermatozoa in the testicular structures may be a prerequisite for the spontaneous production of circulating sperm-agglutinating antibodies, at least of the head-to-tail (H-T) agglutinating type. Furthermore, these circulating H-T sperm-agglutinating antibodies, once they are formed, do not seem to interfere adversely with the germinal epithelium of the carrier.

Agglutination↗
PubMed1983

Sterilization by vas occlusion without transection does not reduce postvasectomy sperm-agglutinating antibodies in serum. A randomized trial of vas occlusion versus vasectomy.

The influence of transection of the vasa on the increase in sperm-agglutinating activity in serum after vasectomy was investigated in a prospective randomized study. Sixty-eight males requesting vasectomy were allocated to either of two groups: vas occlusion by tantalum clips without transection of the vasa, or conventional vasectomy by transection and ligation. Sperm-agglutinating antibodies were determined by a micro-agglutination test before and 6 months after vasectomy or vas occlusion. A significant increase in sperm-agglutinating activity in serum after sterilization was found in both groups, but there was no intergroup difference.

Adult↗
PubMed1982

Sperm-agglutinating and -immobilizing antibody formation following vasectomy prevented with dexamethasone in cynomolgus monkeys.

Cynomolgus monkeys (Macaca fascicularis) were treated with 1.5 mg/kg dexamethasone (DEX) before (4 to 2 days) and after (0, 2, 4, and 7 days) vasectomy. Of the four monkeys treated with DEX, only one developed sperm antibody as measured by sperm-agglutinating and sperm-immobilizing assays. All six of the vasectomized monkeys not given DEX developed both agglutinating and immobilizing sperm antibodies. In this study, DEX given before and after vasectomy blocked sperm-agglutinating and -immobilizing antibody formation. We conclude that the major antigenic exposure to sperm responsible for sperm-agglutinating and -immobilizing antibody comes at the time of vasectomy.

Animals↗
PubMed2003

Sperm granuloma and sperm agglutination in a dog with asthenozoospermia.

A mongrel dog, aged 2 years, was found to have only a small number of sperm, immobilization of all sperm, and many sperm agglutinations in its ejaculates, and scrotal palpation revealed a small nodule in the left cauda epididymis. Addition of the dog's seminal plasma or serum to the semen of 2 normal dogs caused immobilization and agglutination of their sperm. Histological examination showed that the nodule was a sperm granuloma. Many lymphocytes were seen in the stroma around the sperm granuloma. Anti-sperm antibodies are presumed to be present in the semen and serum of the asthenozoospermic dog.

Animals↗
PubMedDate not supplied

Local hydrocortisone treatment of sperm-agglutinating antibodies in infertile women.

Local sperm-agglutinating antibodies (LSAA) in the cervical ovulatory mucus may be a cause of primary infertility. A group of 17 infertile women with LSAA treated without effect with artificial insemination and then condom therapy were studied. After hydrocortisone application to the ectocervix for up to four cycles, LSAA disappeared totally in 13 patients; six of them have given birth to babies. No side effects of treatment were observed. Hydrocortisone for local immunosuppression may become a new method of therapy in cervical immunological infertility.

Administration, Topical↗
PubMed1987

Antibodies to human seminal plasma inhibin cause sperm agglutination and impairment of cervical mucus penetration and sperm-egg attachment.

Inhibin isolated from human seminal plasma which has 94 amino acids has been shown to be structurally similar to a sperm coating antigen of prostatic origin. Specific antibodies generated against this peptide caused agglutination of human sperm. Using FITC-labeled antibody, antigen was localized on the post-acrosomal head region of sperm. Antiserum to inhibin could also impair the penetration of human spermatozoa into cervical mucus. After 10 and 30 minutes, the depth and density of penetration as well as the motility of the sperm were inhibited. The treatment of sperm with antiserum to inhibin caused an inhibition of sperm attachment to the egg as well as inhibition of penetration.

Cervix Mucus↗
PubMed1987

The antibody from an infertile woman that induces sperm agglutination interacts with rat testicular cells and sperm.

The serum obtained from an infertile woman induced a specific head-to-head agglutination of human and rat sperm. The immunoglobulin G (IgG) fraction of the serum was obtained and found to interact with the proteins of rat sperm in testis and epididymis. Using an indirect immunofluorescent method with rat sperm from vas deferens, we determined that the antibody recognized the protein on the convex and concave regions of the acrosome and over the entire tail. However, with testicular spermatozoa, the antibody recognized only the distal end of the tails. In paraffin sections of adult rat testis, sperm tails located at the luminal region of the seminiferous tubules stained intensely. Weak but significant staining also occurred on late spermatids. In the epididymal sections, staining was restricted to spermatozoa in the lumen. On the other hand, sections of testes from 25-day-old rats containing spermatogonia and early spermatocytes had a completely negative reaction. Testicular somatic cells, including Sertoli cells, peritubular myoid cells and interstitial cells, did not stain. To identify the testicular protein interacting with the antibody, adult rat testis proteins were prepared and analyzed by a sodium dodecyl sulfate-polyacrylamide gel electrophoretic (SDS-PAGE) immunoblot technique. The antibody interacted with a protein with an estimated molecular weight of 82,000 in the testicular homogenate and particulate fraction, whereas the reaction was considerably weaker with the testicular cytosol fraction.

Animals↗
PubMed2004

Prostasome antigens as targets for sperm agglutinating antibodies demonstrated by 1-D gel electrophoresis and immunoblottings.

Many immunoinfertile men have sperm agglutinating antibodies that are directed against prostasome-derived antigens, but these antigens have not been defined so far. We selected serum samples with high ELISA titres against prostasomes from a group of immunoinfertile patients with sperm agglutinating antibodies and used the sera for immunoblottings on 1-D SDS-PAGE of prostasomes and sperm cells. The immunoblottings with individual antiprostasome antisera on 1-D SDS-PAGE of prostasomes, revealed three to 10 bands for each serum. Eighty-five per cent of the serum samples contained bands in the 70-75 kDa region and 80% of the samples contained bands in the 50-55 kDa region. Immunoblottings of extracted sperm cells, revealed one to six bands in the molecular weight range 25-82 kDa and two of the samples recognized two bands with molecular weights (50 and 43 kDa) similar to immunoblottings of prostasomes. The prostasomal antigens recognized by the high titre-antisera of immunoinfertile men were generally different from the sperm antigens recognized by the same sera. This suggests that prostasomes offer a new set of antigens available for research on male immunoinfertility and immunocontraception.

Antigens↗
PubMed2006

Contraceptive effect of sperm-agglutinating factor isolated from Staphylococcus aureus in mouse.

OBJECTIVE: To investigate the contraceptive effect of sperm-agglutinating factor (SAF) isolated from Staphylococcus aureus in mouse. DESIGN: Experimental study performed with LACA mice obtained from the Central Animal House, Panjab University, Chandigarh, India. SETTING: In vivo studies conducted in the Department of Microbiology, Panjab University, Chandigarh, India. POPULATION: Sixty female and 18 male mice were used for the studies. METHODS: Mice sperm-S. aureus agglutination, scanning electron microscopy (SEM), in vivo studies in mice. MAIN OUTCOME MEASURE: Contraceptive efficacy of SAF. RESULTS: S. aureus readily adhered to and agglutinated mouse spermatozoa. By SEM, S. aureus adherence was observed on sperm head as well as sperm tail. SAF was found to be causing 100% fertility inhibition in mouse with no effect on vaginal epithelium. CONCLUSIONS: Sperm-agglutinating factor has been found to have a potential as a contraceptive agent.

Agglutinins↗
PubMed1981

Postcoital testing in relation to circulating sperm-agglutinating antibodies in women.

Repeated postcoital tests (PCTs) were evaluated in couples with unexplained infertility, in couples with circulating sperm-agglutinating antibodies of the head-to-head (H-H) or tail-to-tail (T-T) type in serum, and in couples without any antibodies. The results were compared with those of PCTs in the conception cycle from women undergoing gonadotropin treatment for anovulation. Only limited differences in the PCT results were noted between couples with sperm-agglutinating antibodies, couples without such antibodies, and fertile couples.

Anovulation↗
PubMed1982

Hypothalamo--pituitary--gonadal axis and sperm-agglutinating antibodies in infertile men treated for cryptorchidism.

Semen samples, serum levels for luteinizing hormone (LH), follicle stimulating hormone (FSH), prolactin and testosterone, sperm-agglutinating antibodies, and karyotypes were measured in 23 men who had been surgically treated for cryptorchidism and they came to an infertility clinic. Two of these men had normal sperm counts, 2 were azoospermic, and 14 had severe oligozoospermia. Thirteen patients had high FSH levels and two had high LH concentrations. The serum prolactin level was clearly elevated in one man. One man had caryotype 47 XXY. Sperm-agglutinating antibodies were found in three patients.

Adult↗
PubMed2000

Cloning of a glucose phosphate isomerase/neuroleukin-like sperm antigen involved in sperm agglutination.

The mouse monoclonal antibody (mAb) A36 produced by us and shown to induce extensive, "tangled" sperm agglutination was used to isolate cDNAs encoding its cognate antigen. Three overlapping cDNA clones specifically recognized by the mAb were isolated from a human testis cDNA expression library in lambdagt11. Sequencing of these cDNAs yielded the complete nucleotide sequence of a 3-kilobase cDNA that encodes the mAb-related polypeptide, designated sperm antigen-36 (SA-36), composed of 558 deduced amino acids. SA-36 cDNA contained a 5' untranslated region of 234 nucleotides (nt), an open reading frame of 1674 nt, and a 3' untranslated region of 1138 nt. SA-36 cDNA displayed > 99% homology to glucose phosphate isomerase (GPI)/neuroleukin (NLK) mRNA. This surprising homology was confirmed in Western blots demonstrating that mAb A36 reacted specifically with GPI obtained from rabbit muscle and from baker's yeast. Moreover, polyclonal, monospecific antibodies produced against beta-galactosidase/SA-36-3 fusion protein stained human spermatozoa and caused intensive agglutination of these cells in a manner similar to that with the mAb. Taken together, the data presented here demonstrated that mAb A36 cognate sperm surface antigen, encoded by SA-36 cDNA, is a GPI/NLK-like protein involved in sperm agglutination.

Amino Acid Sequence↗