Sperm immobilizing and fertilization-blocking monoclonal antibody 2C6 to human seminal plasma antigen and characterization of the antigen epitope corresponding to the monoclonal antibody.

A monoclonal antibody (Mab 2C6) with strong sperm immobilizing and agglutinating activities was generated by cell fusion between spleen cells from a mouse immunized with human seminal plasma (HSP) and mouse myeloma cells. It also showed a strong inhibitory effect on human sperm-egg interaction. The corresponding antigen was present on the whole surface of ejaculated spermatozoa.

Isolation of four major glycoprotein families (ZP1, ZP2, ZP3, ZP4) of porcine zona pellucida and characterization of antisera raised to each glycoprotein family.

Porcine zona pellucida (ZP) contains antigens which cross-react with man, and anti porcine ZP antibody was found to exhibit a strong blocking effect on human sperm-ZP interaction. Previously, we found that porcine ZP was composed of four major glycoprotein families (ZP1, ZP2, ZP3, ZP4). In this study, each of the glycoprotein families was isolated by O'Farrell's two-dimensional electrophoresis and injected into mice to obtain antisera to each component.

Further characterization of the porcine zona pellucida antigen corresponding to monoclonal antibody 3A4-2G1 exclusively cross-reactive with porcine and human zonae pellucidae.

Biochemical and molecular analyses were performed for the porcine zona pellucida (ZP) antigen corresponding to the monoclonal antibody (Mab) 3A4-2G1 exclusively cross-reactive with porcine and human zonae pellucidae (ZPe). By SDS-PAGE and Western blot analysis, Mab 3A4-2G1 reacted with a 92-kDa component of solubilized porcine ZP (s-PZP) under non-reducing conditions, while reacting with a 23-kDa component under reducing conditions. Diagonal SDS-PAGE revealed that the 92-kDa component was composed of 69 kDa and 23 kDa molecules.

[Coronary revascularization evaluated by thallium-201 myocardial scintigraphy: a follow-up study].

To elucidate changes with time in T1-201 scintigraphy after coronary revascularization, T1-201 stress myocardial scintigraphy was performed at least twice during the follow-up period (from one to 12 months) in 58 patients with ischemic heart disease (12 with angina, and 46 with myocardial infarction) who had undergone PTCA or A-C bypass surgery. The perfusion defects were classified in 4 grades, and scintigraphic changes over grade 1 were judged significant. We evaluated; 1) time of scintigraphic improvement after revascularization, 2) presence of reverse redistribution, and 3) assessment of coronary restenosis.

Correlation between quantitative antibody titers of sperm immobilizing antibodies and pregnancy rates by treatments.

Immunological infertility in women who possessed sperm immobilizing (SI) antibodies made it very difficult to conceive using the usual treatments. We examined SI antibodies by the quantitative Sperm Immobilization Test and found the antibody titers (50% sperm immobilization unit: SI50 unit) associated with pregnancy rates. Patients with high SI50 titers (greater than 10 units) did not conceive by ordinary or repeated artificial inseminations with husband's semen (AIH) except when treated with in vitro fertilization (IVF) and embryo replacement.

Monoclonal antibody recognizing an apparent peptide epitope of human seminal plasma glycoprotein and exhibiting sperm immobilizing activity.

A hybridoma (3B2-F7) has been established which secretes a monoclonal antibody (Mab) directed against a peptide determinant of human seminal plasma glycoprotein (HSP-gP). The deglycosylation of HSP-gP was performed chemically with TFMS hydrolysis and enzymatically in the presence of detergent and further treated with periodic acid after fixing deglycosylated HSP on plastic wells. The Mab 3B2-F7 (IgM, kappa) exhibited sperm immobilization activity (256 units of SI50) and inhibited sperm binding to human zona pellucida.

[Reverse passive hemagglutination assay for human chorionic gonadotropin in urine using monoclonal antibody].

Aiming to find a urinary hCG immuno-assay which is specific, sensitive and easy to perform, a reverse passive hemagglutination reaction was studied by using sheep red blood cells (SRBC) coupled with monoclonal antibodies (Mab) to hCG. Three Mabs (5D4, 6E4, 2F8) with different specialty were used for the study. Mab 5D4 reacted to hCG, hCG-beta, and LH but not to hCG-alpha.

[Experimental study on functional and morphological restoration following hepatic ischemia].

Restoration of damaged hepatocytes after ischemia was studied functionally and morphometrically in a rat model with partial hepatic vascular occlusion. In 30 min occlusion group (n = 35), ATP (mols/mg of protein) dropped down to nearly zero, but returned to the pre-ischemic level within 6 hours after reperfusion. The number of mitochondria (Mt)/unit area of cytoplasm (N/beta m2) increased to 1.

[Factors influencing exercise tolerance in patients with myocardial infarction as elucidated by Tl-201 myocardial scintigraphy].

Measurements of O2 consumption during treadmill exercise tests and Tl-201 myocardial scintigraphy were performed in 17 cases of myocardial infarction to elucidate O2 consumption at an anaerobic threshold ATVO2 as an adequate index of exercise tolerance, and the scintigraphic indices influencing the exercise tolerance. ATVO2 was obtained using the method of Wasserman and Davisand corrected by body weight. The scintigraphic indices such as the location, extent (residual myocardium), and severity (% uptake) of myocardial infarction were obtained from SPECT and bull's eye displays on Tl-201 myocardial scintigraphy.

Techniques for sperm immobilization test.

Several semiquantitative, quantitative, and microassay techniques had been developed to detect antibodies bound to human spermatozoa: sperm agglutination test (SAT), sperm immobilization test (SIT), immunofluorescence test, radioantiglobulin test, enzyme-linked immunosorbent assay (ELISA), mixed erythrocyte-spermatozoa antiglobulin reaction (MAR), "Panning" test, and immunobead test (IBT). Clinical application of these techniques include (a) detection of sperm immobilizing antibodies in sera of sterile women, (b) follow-up study of sperm immobilizing antibodies, and (c) detection of sperm immobilizing antibodies in cervical mucus and other secretions. The chemical structure of antigen epitope corresponding to Mab H6-3C4 may recognize the internally located repetitive unbranched N-acetyllactosamine structure, regardless of terminal substitution at Gal (i.

Characterization of epitopes of seminal plasma antigen stimulating human monoclonal sperm-immobilizing antibodies: a personal review.

In observations made between 1974 and 1984, 40 of 303 women with unexplained sterility (13.2%) showed positive sperm-immobilizing antibodies. Among many kinds of antibodies to human sperm including sperm coating antigens, the biologically active antibodies, such as sperm-immobilizing agglutinating antibodies and blocking antibodies for fertilization, could be relevant to infertility.

Characterization of a small immunoreactive HCG-like component isolated from urine of a patient with choriocarcinoma.

An immunoreactive hCG-like substance with low molecular weight was isolated from the urine of a patient with choriocarcinoma and its characteristic properties were studied. The urinary hCG preparation was fractionated by gel filtration on a Sephadex G75 column and each fraction was assayed for hCG activities by an enzyme linked immunosorbent assay using a monoclonal antibody to hCG (Mab-5D4). Two immunoreactive hCG peaks were obtained.

Immunogenicity of a 92kDa component of porcine zona pellucida isolated using a monoclonal antibody (3A4-2G1) exclusively cross-reactive with porcine and human zonae pellucidae.

Three monoclonal antibodies (Mabs) to solubilized porcine zona pellucida (s-PZP) were produced. They reacted exclusively to porcine and human zonae pellucidae (ZPe) and two of them blocked sperm penetration of ZP in humans by collaborating with goat anti-mouse gamma-globulin serum as a second antibody. The antigen corresponding to one of these Mabs (Mab 3A4-2G1) was fractionated from s-PZP by immunoaffinity chromatography bound Mab 3A4-2G1.

[Monoclonal antibodies].

Since the first successful application of somatic cell fusion by Kohler and Milstein to generate hybridomas secreting monoclonal antibodies (Mabs), this hybridoma technique has been widely applied for production of Mabs against a wide variety of antigens. In the lecture, general procedures for establishment of hybridomas and characteristic properties of Mabs were briefly explained and followed by presentation of our clinical studies using Mabs produced in our laboratory to hCG, sperm and tumor cells. 1.

Application of the quantitative sperm immobilization test for follow-up study of sperm-immobilizing antibody in the sera of sterile women.

For quantitative estimation of sperm immobilizing antibody, a new assay method was developed and applied to a follow-up study of antibody titers in the sera of sterile women. The 50% sperm immobilization units (SI50) estimated by the quantitative method ranged from 1.2 to 97.

[Effect of immunosuppressants on the subrenal capsule (SRC) assay as a chemosensitivity test].

Six-day SRC assay as a chemosensitivity test has an advantage of high predictive rate for clinical response. However, it is pointed out that very few viable tumor cells are observed at the end of the assay, so that it may make the assay results unreliable. In this paper, we tested the effect of immunosuppressants on SRC assay using Walker carcinosarcoma originated from Wistar rat xenografted under the renal capsule of BDF1 mice.

Analyses of epididymal sperm surface antigens by monoclonal antibodies against hamster spermatozoa.

BALB/c mice were immunized with spermatozoa from cauda epididymides of hamsters and the immune spleen cells were fused with mouse myeloma cells (P3U1). Seven hybridomas (GHS-1,-2,-3,-4,-5,-6, and -7) that produced monoclonal antibodies (Mabs) binding to the epididymal spermatozoa were established. Three Mabs (GHS-3,-4, and -6) were IgM and the other four were IgG1.

Establishment of a murine monoclonal antibody against human clear cell carcinoma and analysis of the corresponding antigen.

A murine monoclonal antibody (Mab) 4B6, immunoglobulin M (IgM), lambda chain, against human clear cell carcinoma of the ovary was established. Mab 4B6 reacted specifically to clear cell carcinomas, but failed to react to other types of ovarian carcinomas, such as mucinous, serous carcinoma, and also failed to react to human normal organ tissues. Mab 4B6 recognized antigenic determinants located on the membranes of carcinoma cells.

[Successful application of in vitro fertilization and embryo replacement in the treatment of infertile women with sperm immobilizing antibody].

Thirteen infertile patients (18 cycles) with sperm immobilizing antibodies were subjected to in vitro fertilization and embryo replacement (IVF-ER) therapy during 12 months from January to December 1985. Four patients became pregnant, two of them delivered healthy babies at term and 2 had abortions at 5 weeks and 12 weeks of gestation, respectively. In the same duration, 24 patients (28 cycles) with tubal factor and 4 patients (5 cycles) with male factor for infertility were subjected to IVF-ER, and two patients with tubal factor became pregnant and delivered healthy babies at term.