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S Isojima

Publications and source records attributed to S Isojima.

At least 73 records · Page 4Linked to original sources
PubMed1985

Identification of two different surface epitopes of human ovarian epithelial carcinomas by monoclonal antibodies.

Monoclonal antibodies (Mabs) against human ovarian mucinous and serous carcinoma were obtained by the Mab technique. Their reactivities against human tumors were tested by indirect immunofluorescence. One of the Mabs, named 4C7, derived from the spleen cells of mice immunized with mucinous carcinoma line OVA-1, reacted to ovarian mucinous carcinoma, endometrioid carcinoma, and mesonephroid carcinoma but did not react to ovarian serous carcinoma. Another Mab, 3C2, obtained from the spleen cells of mice immunized with serous carcinoma line HOC-21, reacted to serous carcinoma and endometrioid carcinoma of ovary, but never reacted to mucinous carcinoma or mesonephroid carcinoma. Neither of the Mabs reacted to other types of ovarian carcinomas such as undifferentiated carcinoma, dysgerminoma, endodermal sinus tumor, and malignant teratoma of ovary, and also did not react to any benign ovarian tumors or other normal human tissues. Both Mabs 3C2 and 4C7 had no reactivity to carcinoma of other organs such as stomach, colon, lung, lymphoid system, and kidney and also did not react to human lymphocytic or carcinoembryonic antigen as confirmed by using many human cell lines and surgically resected samples. Since the cross-reactivities of these Mabs were limited within the ovarian epithelial carcinomas, it is suggested that two distinct epitopes are expressed on the ovarian epithelial carcinomas. One epitope, identified by Mab 4C7, is expressed only on mucinous carcinoma, endometrioid carcinoma, and mesonephroid carcinoma, while the epitope, identified by Mab 3C2, appears only on serous and endometrioid carcinoma.

Adenocarcinoma, Mucinous↗
PubMed1985

[Production and characterization of a monoclonal sperm immobilizing antibody to seminal plasma antigens].

Rat spleen cells immunized to human azoospermic semen were fused with mouse myeloma cells (P3 X 63Ag8Ul) to obtain hybridomas which produce monoclonal sperm immobilizing antibodies. Of 114 hybrid clones obtained, 10 were found to produce antibodies with sperm immobilizing activity with complement. One clone (1C4) producing the highest antibody titer was chosen for further characterization. Chromosome analysis showed that 1C4 hybridoma cells contained two types of chromosomes which were derived from rat and mouse. Rat Ig was detected in both the cytoplasma and the culture medium of the cells by direct immunofluorescent test and immunodiffusion test, respectively. Gammaglobulin fractions from the culture medium had a strong sperm immobilizing activity and showed a common single precipitin line against human seminal plasma and human milk proteins. It was proved that the monoclonal antibody was produced to No. 7 antigen and not to No. 3 antigen (lactoferrin) in human seminal plasma, both of which have a common antigenicity to human milk proteins and compose the sperm coating antigens.

Animals↗
PubMed1984

Monoclonal antibodies to porcine zona pellucida antigens and their inhibitory effects on fertilization.

Five hybridomas which produce monoclonal antibodies (Mabs) to porcine zona pellucida (ZP) were established. The immunoglobulin classes were IgG2a from hybridoma B11C8 and IgM from the other four. Using immunofluorescent staining, the Mab from B11C8 stained only porcine oocytes, the Mab from G10G5 stained oocytes of pigs and hamsters but Mabs from C6H1, D3H4, G10F9 stained oocytes of pigs, humans, hamsters, rats and mice. Mabs from B11C8 and G10F9 strongly blocked boar sperm binding to porcine ZP but other Mabs produced only slight blocking. However, no Mabs could block sperm penetration during in vitro fertilization (IVF) of hamster oocytes. When a goat antibody (IgG) to mouse serum gamma-globulin was applied after each Mab as a second antibody, only Mab from G10G5 impaired IVF of hamster oocytes.

Animals↗
PubMed1984

[Detection of early pregnancy factor in the sera of conceived women before nidation].

In order to apply the early pregnancy factor (EPF) to early diagnosis of fertilization, the establishment of optimal conditions for assay of EPF was attempted, and then EPF in the sera of contracepted and conceived women 4 to 6 days after ovulation were measured. For assay of EPF, 0.25 ml of 1:2 step diluted anti-human lymphocyte serum (ALS), 0.05ml of guinea pig serum as complement and 0.1ml of lymphocytes suspension (1 X 10(7)/ml) pretreated with test serum were mixed and then incubated at 37 degrees C for 90 min. To this mixture 0.1ml of sheep red blood cell suspension (2 X 10(9)/ml) was added and the rosette formation was counted after centrifugation. The rosette inhibition titer (RIT) was expressed as reciprocal of ALS dilutions which resulted in less than 75% of rosette formation as compared with the control. RITs of the conceived women who were assayed on the 5 th day after ovulation were in the range from 16 to 32 X 10(3), while that of the control contraceptive women who were assayed on the same day was in the range from 2 to 4 X 10(3). The sterile women who received AIH but failed to conceive all showed less than 4 X 10(3) as RIT. These results suggest that the assay of EPF is valuable in detecting the early stage of fertilization and possibly may help to differentiate the impairment of embryo implantation from non-fertilization of the ovum as a cause of sterility.

Chaperonin 10↗
PubMed1983

Localization of human seminal plasma No. 7 antigen (Ferrisplan) in accessory glands of male genital tract and spermatozoa.

The establishment of a hybridoma (1C4) producing sperm immobilizing monoclonal antibody to human seminal plasma No. 7 antigen (HSP No. 7 Ag.) and the isolation of the pure antigen by immunoaffinity chromatography bound monoclonal antibodies have been reported previously. In the present investigation, HSP No. 7 Ag. has been termed 'Ferrisplan' and its distribution in male genital organs, spermatozoa and body fluids has been studied. The amount of Ferrisplan in the body fluids was determined by radioimmunoassay. Large amounts were detected in seminal plasma and milk, trace amounts in saliva, and none in the serum and urine. The concentration of Ferrisplan was highest in the seminal plasma of azoospermic patients and gradually decreased from oligospermia to normospermia. Using an immunofluorescent method with anti-Ferrisplan monoclonal antibody, strong staining was observed on the epithelial layers of human seminal vesicles, no staining on testes and bright staining on the post-nuclear cap and mid-piece segment of spermatozoa. These results indicate that Ferrisplan is excreted mainly from the seminal vesicle and adheres to the post-nuclear cap and mid-piece of the spermatozoa as a sperm-coating antigen.

Antigens↗
PubMed1982

Purification of human seminal plasma no. 7 antigen by immunoaffinity chromatography on bound monoclonal antibody.

Human seminal plasma (HSP) No. 7 antigen was purified by immunoaffinity chromatography on bound 1C4 monoclonal antibody (Moab) (Shigeta et al., 1980b). The pooled HSP protein was applied to a CNBr-activated Sepharose 4B column of bound 1C4 Moab gamma globulin and the antibody bound fraction (fr) eluted was further purified by rechromatography in the same way. The purified antigen in the antibody bound fr obtained by rechromatography gave a single band on SDS-PAGE in a position corresponding to a molecular weight of 15,000 daltons. This preparation was 196.2 times more effective than the original HSP protein in neutralizing the sperm immobilizing activity of 1C4 Moab. The purified HSP No. 7 antigen contained iron, but was different from lactoferrin and transferrin. It did not show any enzymatic activities, such as those of acid phosphatase, LDH or trypsin inhibitor, and shared antigenicity with human milk protein. It was present in seminal plasma as a molecule with a higher molecular weight but seemed to be cleaved to a monomer of 15,000 daltons during purification procedures. This antigen is present on spermatozoa as sperm-coating antigen and the corresponding antibody can immobilize spermatozoa with complement.

Antibodies, Monoclonal↗
PubMed1981

Effect of purified zona antibody on fertilization in the mouse.

Bivalent (IgG) rabbit anti-mouse zona pellucida antibody completely inhibited fertilization in the mouse. Univalent (Fab) zona antibody did not inhibit the fertilizability of eggs. However, the treatment or injection with anti-rabbit IgG serum inhibited the fertilization of eggs pretreated with antiserum Fab or eggs from females injected with antiserum Fab. It is concluded that the inhibition of fertilization by bivalent louse zona antibody depends on the possibility that the antisera are directed against the zona as a whole and not specifically against the receptors for the sperm.

Animals↗
PubMed1981

[Role of complement for the sensitivity of sperm immobilization test (author's transl)].

For detecting the sperm immobilizing antibody in the sera of infertile women with unknown cause, Isojima's sperm immobilization test was widely used, but there was great difference on the percentage of positive results among several laboratories. For analysis of factors influencing the sensitivity of the test, source and amount of complement were studied, and the following results were obtained. 1. Guinea pig serum which contained more than 200 CH50 as hemolytic titer was best for this test. 2. Rat and rabbit sera which contained much lower CH50 than guinea pig indicated relatively good sensitivity for the test. 3. Human serum which contained 34.9 CH50/ml and mouse serum which contained less than 5 CH50/ml, both could not work on sperm immobilization test. The human serum (AB) which was concentrated to the level of guinea pigs serum as CH50 did not work well on sperm immobilization test as guinea pig serum. 4. When the complement from same source was used, the positive correlation between CH50 (hemolytic titer) and 50% sperm immobilization titer (SI50) was present, but if the complement source was different, there was no correlation between CH50 and SI50. 5. It was proved that more than 10 CH50 of guinea pig serum was necessary for sperm immobilization test to obtain the most sensitive results.

Animals↗
PubMed1981

[Studies of fertilizing capacity of spermatozoa from infertile men with oligozoospermia. AIH by washed and concentrated spermatozoa and in vitro fertilizing capacity to zona free hamster eggs (author's transl)].

The fertilizing abilities of spermatozoa from infertile men with oligozoospermia were evaluated by examining pregnancy rates after AIH using washed and concentrated spermatozoa, and also by in vitro fertilizing rates to zona free hamster eggs. The spermatozoa from ten oligozoospermic men with less than 20 x 10(6)/ml of sperm counts were used in the experiments. After the procedures of washing and concentrating spermatozoa from oligozoospermic men, the sperm concentration was increased from two to six times and the sperm motility was also increased up to two times. However none of the patients' wives conceived after several AIHs with washed and concentrated spermatozoa. When the in vitro fertilization of zona free hamster eggs by the spermatozoa was used, the fertilization rates were null in case of 7 oligozoospermic patients, and 5, 10 and 33.3% in case of another 3 oligozoospermic patients respectively in contrast with 73.3% in use of spermatozoa from control fertile men. The fertilizing abilities of spermatozoa from husbands of 4 sterile couples with an unknown cause were examined by in vitro fertilization test and 7.1, 37.5, 50.0 and 55.6% fertilization rates were obtained. Among these men, the spermatozoa from one man indicated a very low fertilization rate without any other pathological findings. This suggests that the impairment of fertilizing ability of spermatozoa might be present in the case of sterile men with an unknown cause.

Adult↗
PubMed1981

[Comparative studies on doses of complement for detection of sperm immobilizing antibody in serum, cervical mucus and seminal plasma (author's transl)].

In Isojima's sperm immobilization test, the complement is one of the most important factors which influence on sensitivity of the test. Previously we reported that more than 10 CH50 of guinea pig serum complement was necessary in the reaction to obtain the best result for measuring the sperm immobilizing antibody in the serum. This time, the influences of various test specimens on complement activities for the sperm immobilization test were studied and the necessary amounts of complement to detect the sperm immobilizing antibody in serum, cervical mucus and seminal plasma were compared. In the sperm immobilization test, the following results were obtained: 1. The amount of complement (guinea pig serum) added was 11.9 CH50, and after 60 minutes incubation at 32 degrees C in veronal buffer (VB2+), 2.0 +/- 0.3 (M +/- SD) of CH50 was consumed. 2. When human serum, cervical mucus and seminal plasma were added to the reaction as specimens, the complement consumptions (M +/- SD) were more 0.2 +/- 0.3 CH50, 0.2 +/- 0.2 CH50 and 3.3 +/- 0.8 CH50 than that of VB2+ buffer respectively. 3. When the washed human spermatozoa were added, the consumed amounts of complement (M +/- SD) were dose dependent; 3.5 +/- 0.7 CH50 for 100 X 10(4) spermatozoa, 7.3 +/- 0.3 CH50 for 200 X 10(4) spermatozoa and 15.7 +/- 2.2 CH50 for 400 X 10(4) spermatozoa. 4. In our sperm immobilization test, the net consumptions of complement (M +/- SD) were 5.0 +/- 0.7 CH50 for specimen of serum, 4.9 +/- 0.8 CH50 for specimen of cervical mucus and 11.1 +/- 1.2 CH50 for specimen of seminal plasma. 5. Complement doses necessary to obtain the most sensitive sperm immobilization tests were 10 CH50 for both serum and cervical mucus specimens, and 15 CH50 for seminal plasma specimen.

Animals↗
PubMed1980

Sperm-immobilizing monoclonal antibody to human seminal plasma antigens.

Rat spleen cells immunized to human azoospermic semen (a mixture of seminal plasma components) and mouse myeloma cells (P3/X63 Ag8U1; P3U1) (Marguilies et al., 1976) were successfully fused with polyethylene glycol (PEG 1500) and 19 of 89 fused cell cultures were found to produce sperm-immobilizing antibody. The cells that produced antibody indicating the highest sperm-immobilizing activity were distributed into wells for further recloning and 10 clones producing sperm-immobilizing antibody were established. The clone (1C4) producing the highest antibody titre was found to produce a large amount of IgG in culture supernatants and to contain a mixture of rat and mouse chromosomes. It was proved by immunodiffusion test that the monoclonal antibody was produced to the human seminal plasma antigen No. 7 which is common to human milk protein. Using this hybridoma which produced a large amount of monoclonal sperm-immobilizing antibody, a new method could be developed for purifying human seminal plasma antigen by immunoaffinity chromatography with bound antibody from the hybridoma.

Animals↗