Search PubMed⌕ Search

Biomedical subjects

A C Menge

Publications and source records attributed to A C Menge.

At least 55 records · Page 3Linked to original sources
PubMed1984

Isoantigenicity of rabbit sperm, testis, and their extracts as demonstrated by Western blot enzyme immunobinding procedure.

The isoantigenicity of rabbit germ cells and their extracts was studied by using a Western blot enzyme immunobinding procedure after systemic and local intrauterine immunization. In the deoxycholate (DOC) sperm extract, the antisperm isoantiserum (ASA) recognized proteins of 100, 96, 92, 82, 53, 47, 43, and 29 kD, and the antitestis isoantiserum (ATA) recognized proteins of 100, 96, 92, 82, 43, and 29 kD. In the lithium diiodosalicylate (LIS) sperm extract, ASA reacted with four antigen bands of 92, 30, 14, and 6 kD, and ATA reacted with proteins of 94, 92, 91, 23, 17, 7, and 6 kD. In the reduced sodium dodecyl sulfate (SDS) sperm extract, ASA recognized proteins of 95, 76, 51, 41, 33, 30, 28, 26, and 22 kD; ATA showed a reaction only with proteins of 100, 98, and 95 kD, and anti-LIS/sperm isoantiserum (ALA) recognized three proteins of 98, 81, and 65 kD. The IgA of the immune uterine fluid (IUF) from the LIS/sperm-immunized rabbits recognized a 14-kD antigen in the DOC conceptus extract, a 51-kD antigen in the DOC testis extract, and 53-, 49-, 31-, and 28-kD proteins in the reduced SDS-sperm pellet extract. The IgA of the IUF from the NP-40/pellet-immunized rabbits showed two proteins of about 100 kD, plus ones of 56, 36, 31, 29, 19, 17, and 14 kD in the DOC conceptus extract; proteins of 92, 70, 51, 29, and 14 kD in the DOC testis extract; and of 61, 57, 53, 49, 35, 31, and 28 kD in the reduced SDS-sperm pellet extract.

Animals↗
PubMed1984

Monoclonal antibodies to rabbit sperm autoantigens.

Rabbits were immunized with homologous spermatozoa to investigate their autoimmunogenic nature. Major sperm autoantigens that elicit antisperm antibodies were analyzed molecularly by the sodium dodecylsulfate (SDS) gel/protein blot radioimmunobinding method. IgG fractions of the autoimmune sera were purified by a protein A-Sepharose column, immobilized on Sepharose as affinity ligands, and utilized to purify major sperm autoantigens from rabbit testes. The autoimmunogenicity of the purified autoantigens was verified by reimmunizations in rabbits. BALB/c mice were immunized with the affinity-purified autoantigens to raise monoclonal antibodies by modified hybridoma techniques. Following fusions and clonings, we have established more than 100 hybrid cell lines that were shown to secrete antibodies to purified autoantigens and to rabbit sperm. A variety of techniques was employed to characterize these monoclonal antibodies. By the SDS gel/protein blot radioimmunobinding method, some were found to react with unique proteins of rabbit spermatozoa. By indirect immunofluorescent assay, about one third were shown to bind different cytologic regions of spermatozoa from rabbit, man, and/or mouse. Six were found to inhibit rabbit sperm binding to rat ova in vitro. In addition, agglutinating and immobilizing activities of these antibodies on live sperm were observed.

Animals↗
PubMed1984

Effect of antisperm antibodies in serum and cervical mucus on the capacity of human sperm to penetrate zona-free hamster ova.

Human sera with elevated titers of antisperm antibodies reduced the capacity of human sperm to penetrate zona-free hamster ova. Of 111 sera examined, 33 (30%) reduced the mean percentage of ova penetrated to less than 52%, which was significantly different from the overall control mean (75%). Three sera reduced the penetration of ova to 7% or less. The inhibitory effect of these three sera was still present in their respective Fab preparations. An analysis of antibody effects revealed that the presence of sperm-immobilizing antibodies had a significantly greater influence on sperm penetration capacity than agglutinating antibodies. Sperm motility per se appeared to be unaffected by the different sera. Sperm treated with cervical mucus (CM) samples generally exhibited a lesser degree of ova penetration than did sperm treated with serum or medium alone. Among CM groups, however, samples containing detectable sperm-immobilizing antibody activity or samples from women with circulating antisperm antibody caused significant reductions in sperm penetration capacity.

Animals↗
PubMed1983

Inhibition of fertility in rabbits by monoclonal antibodies against sperm.

Monoclonal antibodies were prepared against rabbit sperm antigens by fusing P3-X63-Ag8-653 mouse myeloma cells with lymphocytes from Balb/c mice immunized with Tergitol NP-40 detergent-solubilized rabbit epididymal sperm. Ascites fluid from mice injected with two of these hybridomas (8C4.1 and 8C10.5) was negative in immobilization and agglutination methods, however, acrosome positive on methanol fixed sperm and plasma membrane positive on unfixed sperm in indirect immunofluorescence. Insemination of female rabbits with the sperm treated with either of these monoclonal antibodies resulted in significant reduction in fertility as seen by the percentage of 9-day implants/corpora lutea ratio (8C4.1, 25.7%; 8C10.5, 1.9%; and control, 64.7%). Though the antibodies inhibited in vitro binding of the rabbit sperm with zonae pellucidae of rat ova, fertilization in vivo was not affected significantly. The antibodies did not demonstrate antiblastocyst activity by immunofluorescence. Both of these monoclonal antibodies appeared to recognize the same antigen by the SDS-PAGE/Protein Blot enzyme immunobinding procedure. The antigen was of testicular origin and had a molecular weight of approximately 63,000 daltons. It is concluded that these monoclonal antibodies which were organ specific, block post-fertilization fertility by inhibiting some step necessary for viable embryo formation.

Animals↗
PubMed1983

Identification of human sperm antigens to antisperm antibodies.

We have successfully applied SDS (sodium dodecyl sulfate) gel/protein blot radioimmunobinding method to identify the molecular size of sperm antigens that elicit antisperm antibodies from patients with unexplained infertility. Following the transfer of renatured proteins from SDS gel of human sperm extract onto nitrocellulose strips, the radioimmunobinding was performed by incubating the strips with patients' sera at 1:100 dilution and then with I125-labeled goat antihuman immunoglobulin G (IgG) or protein A as detecting probes. Unique sperm antigens that reacted with some patients' sera were identified following the autoradiography of the incubated paper strips. Among the fifty-nine standard serum samples from the Reference Bank of the World Health Organization, about one-fourth of them were found to react predominantly with a sperm protein band having the reference value (Rf value) of 0.2 and the approximate molecular weight of 90,000 dalton. A similar analysis was also performed with serum samples from vasectomized patients. Some of them also revealed a specific binding with the sperm antigen(s) of similar molecular weight. The results of this analysis were also compared with those of conventional tests for sperm antibodies as well as those of microplate radioimmunoassays and enzyme-linked immunoassays. This study suggests that SDS gel/protein blot radioimmunobinding method can be a useful tool for the molecular identification of unique human sperm antigen(s) that elicit naturally occurring antisperm antibodies in patients with unexplained infertility.

Antibodies↗
PubMed1982

Monoclonal antibodies to human sperm antigens.

To elucidate the molecular nature of human sperm autoantigens, attempts were made to raise monoclonal antibodies against these antigens, by hybridoma techniques. After successive immunizations with the particulate fractions of human sperm extract in BALB/c mice, the spleen cells were fused with P3-X63-Ag8 myeloma cells. Several clones and their subclones were obtained and shown by microplate radioimmunoassay to produce antibodies against human sperm antigens. When SDS gel/protein blot radioimmunobinding was used for further molecular analysis, three independently derived clones were shown to produce antibodies, all of which cross-reacted with the same two human sperm antigens with a molecular weight of about 10,000. Using an indirect immunofluorescence assay, antibodies produced by these clones were shown to react with antigens localized on the acrosomal regions of human spermatozoa. Monoclonal antibodies produced by other clones, however, showed no cross-reactivity with any of the blotted proteins from SDS gels of human spermatozoa. Some possible reasons for this are presented.

Animals↗
PubMed1982

The incidence and influence of antisperm antibodies in infertile human couples on sperm-cervical mucus interactions and subsequent fertility.

Analysis of serum samples from 698 infertile couples revealed antisperm antibodies present in 16.5% of the men and 21.6% of the women. Overall, 31.1% of the couples possessed at least one individual with positive results. Sperm-immobilized activity was detected in 29.6% of the cervical mucus (CM) samples from 459 women. Reduced sperm penetration of CM was significantly associated with serum titers of antisperm antibodies in both sexes and also with immobilizing activity in CM of women. The incidence of subsequent pregnancy in 376 infertile couples was reduced significantly if one or both partners had antisperm antibodies in serum or in genital tract secretions. The latter was reflected by evaluation of the immobilization, penetration, and shaking phenomenon of sperm in CM.

Antibodies↗
PubMed1981

Studies on the use of estrous bovine cervical mucus in the human sperm-cervical mucus penetration technique.

Estrous bovine cervical mucus (BCM) was evaluated as a practical substitute for midcycle human cervical mucus (HCM) in the sperm-cervical mucus penetration technique (S-CMPT). Analysis of semen samples from 299 men of infertile couples suggested that up to 95% of the S-CMPT analyses with BCM were compatible with results expected with HCM. Results from a secondary S-CMPT with BCM using donor sperm treated with seminal plasma or cervical mucus extracts from partners of infertile couples indicated 70 and 88% agreement with the clinical S-CMPT obtained with the semen and cervical mucus samples, respectively. Seminal plasma samples from men with serum antisperm antibodies decreased the ability of donor sperm to penetrate BCM in the Secondary S-CMPT. Our results indicate that estrous BCM possess those basic qualities of HCM that support human sperm motility, allow for normal sperm penetration, and interact with antibody-treated sperm to inhibit penetration and to produce the sperm shaking phenomenon.

Agglutination↗
PubMed1980

Antibody activities of serum and uterine fluid samples from rabbits isoimmunized against sperm fractions.

Sperm and uterine fluid samples were collected from rabbits that had been isoimmunized systemically and were challenged locally against a sperm extract (soluble fraction of lithium diiodosalicylate extraction) and a sperm-pellet extract (soluble fraction of Nonidet NP.40 treatment of pellet remaining after LIS extraction). Antibody activity against the sperm plasma membrane was found in serum and uterine fluid samples from the sperm-extract immune rabbits, as indicated by an inhibition of sperm attachment to ova in vitro and by indirect immunofluorescent staining on unfixed sperm. A strong acrosome staining was noted with fixed sperm. These samples did not show fluorescent staining of swollen heads of sperm or react with blastocysts. The immune samples from rabbits injected with sperm-pellet extract displayed the opposite reactions. These samples did not react with the plasma membrane of sperm but the IgA antibodies of the uterine fluids caused immunofluorescent staining of the swollen heads of sperm and the blastocyst cell membranes. Uterine fluid from this group of rabbits also inhibited blastocyst development in vitro. Immune samples from rabbits immunized against untreated epididymal sperm gave all the positive reactions.

Animals↗
PubMed1979

Effects of antisera on human sperm penetration of zona-free hamster ova.

Human sperm, after treatment with a 10% concentration of rabbit or rhesus monkey normal sera and antisera, were evaluated for fertilizing potential by incidence of zona-free hamster ova penetrated by the sperm as evidenced by the presence of swollen sperm heads and male pronuclei. Compared with the basic medium alone, treating sperm with normal sera tended to increase the percentage of ova penetrated whereas antisera against sperm, sperm extract, and testis caused significant decreases in ova penetrated. A Fab preparation of these antisera exhibited similar inhibitory effects. Antisera Fab treatment of the zona-free ova prior to exposure to sperm had no effect on penetration rate.

Animals↗
PubMed1975

Evaluation of human sera for antibodies against sperm by immunofluorescence.

Newborn cord sera and sera from pregnant women and from couples with unexplained infertility were observed by indirect immunofluorescence for reactions against human sperm. The sera from the infertile couples had been tested for macro-agglutination and complement-dependent immobilizing antibodies as well. Nonspecific fluorescence was noted only on the quatorial segment. Acrosomes and tail end-pieces were strongly immunofluorescent (positive at a serum dilution of 1:16) with 60 to 70% and 30 to 50%, respectively, of the sera from each main group, with the exception of the nonreactive cord blood. End-piece fluorescence was primarily due to the IgM globulin fraction. It is suggested that cord serum anti-sperm antibodies, directed mainly against the tail main piece, are due to IgG passively acquired from the mother. There was no apparent difference in immunofluorescence between infertile patients and control groups (except for the acrosome and end-piece, with cord sera). However, when the sera from infertile patients were subdivided into groups containing only agglutinating antibody, only immobilizing antibody, and both types of antibody, a higher percentage of the sera from the group having only agglutinating antibody reacted with all sperm areas. This reaction seemed more uniform over the sperm and included the usually negative posnuclear cap and midpiece. The other subgroups showed no obvious trends.

Acrosome↗
PubMed1975

Testis antigens of man and some other primates.

Rabbit antisera raised aginst testis preparations of human, chimpanzee, rhesus monkey, and baboon origin were used to study testis-specific antigens within and among the four primate species. Antisera were absorbed with serum, liver, kidney, and spleen preparations of the respective species against which they had been produced. Immunoelectrophoretic analysis of testis extracts, using the absorbed antisera, indicated the following minimum numbers of testis-specific antigens for each species: man, 10; chimpanzee, 8; rhesus monkey, 10; and baboon, 8. Most of the testis antigens were cross-reactive among species. The results suggest that human spermatozoa possess at least four to five specific antigens which originate in the testis. The antigen that induces sperm-immobilizing antibody cross-reacted among the four species of primates. Human and rhesus monkey sperm reacted equally in the immobilization system. Testis extracts from each primate species were capable of removing the sperm-immobilizing activity of human immune sera by absorption. Testis proteinase activity, as determined by using a gelatin membrane substrate, was inhibited by the gamma-glogulin fractions of rabbit and rhesus monkey antisera and also appeared to be cross-reactive among the species.

Animals↗