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Biomedical subjects

T Kajino

Publications and source records attributed to T Kajino.

At least 37 records · Page 2Linked to original sources

Is immunotherapy for habitual aborters an immunologically hazardous procedure for infants?

Physical development and tests of immunologic function are reported from the first year of life for 13 infants born to mothers who were habitual aborters and who had undergone subcutaneous vaccination with their husband's lymphocytes. The mean weight of the infants at birth was 2,975 +/- 540 g, including one infant who was small for dates. Physical development parameters for the first year were all within normal range. Immunologic studies were performed at ages 0, 1, 3, 6, 9, and 12 months. The studies included the following: (1) serum levels of immunoglobulins and complement components; (2) the number of peripheral blood lymphocytes (PBL) and their subpopulations; and (3) proliferative responses by PBL against unrelated lymphocytes and some mitogens (pokeweed, phytohemagglutinin, and concanavalin A). All were normal when compared with studies of infants of nonimmunized mothers. These observations suggested that subcutaneous vaccination of women with their husband's lymphocytes did not result in any adverse effects on their infants' physical or immunologic development.

Abortion, Habitual↗

Structure of a Ruminococcus albus endo-1,4-beta-glucanase gene.

A chromosomal DNA fragment encoding an endo-1,4-beta-glucanase I (Eg I) gene from Ruminococcus albus cloned and expressed in Escherichia coli with pUC18 was fully sequenced by the dideoxy-chain termination method. The sequence contained a consensus promoter sequence and a structural amino acid sequence. The initial 43 amino acids of the protein were deduced to be a signal sequence, since they are missing in the mature protein (Eg I). High homology was found when the amino acid sequence of the Eg I was compared with that of endoglucanase E from Clostridium thermocellum. Codon usage of the gene was not biased. These results suggested that the properties of the Eg I gene from R. albus was specified from the known beta-glucanase genes of the other organisms.

Amino Acid Sequence↗

Antibodies to trophoblast in normal pregnant and secondary aborting women.

Enzyme-linked immunosorbent assays (ELISA) with the use of chorionic villous plasma membranes prepared from first trimester and term placentae were employed to detect antibodies to trophoblast in normal primigravid women. Normal pregnant women were found to produce IgG antibodies to trophoblast. These antibodies could be eluted from first trimester placentae. This antibody response was observed in the first trimester and gradually decreased as pregnancy progressed. IgM antibody responses were observed only in the third trimester. Antibodies in some primigravid women and secondary recurrent aborters showed allotypic reactivity with individual trophoblast membranes. This finding was confirmed by immunoblotting experiments in which antibodies from some normal pregnant women were shown to recognize the same trophoblast antigens as those recognized by antibodies from secondary recurrent spontaneous aborters.

Abortion, Habitual↗

Trophoblast antigens in human seminal plasma.

Secondary recurrent spontaneous (2 degrees) aborters manifest persistent IgG, which show differential cytotoxicity patterns with lymphocytes from many donors. These are non-HLA-directed antibodies, which react allotypically with both trophoblast and lymphocytes. The antigens they recognize are designated trophoblast-lymphocyte crossreactive (TLX) antigens. Xenogeneic anti-TLX sera were studied with the use of enzyme-linked-immunosorbent (ELISA) and immunochemical assays to determine the TLX status of seminal plasma. The results showed 1) allotypic TLX antigens are present in seminal plasma; 2) seminal plasma TLX antigens may be membrane associated; 3) by immunoblotting, the molecular weights of antigens reactive with TLX antisera are 15, 22, 28, 33 kD and a smear between 180 and 340 kd; 4) by isoelectric focusing, TLX antigens show pI 4.0, 5.35, 5.9, 6.5, 6.8, and 7.2. Allotypic seminal plasma TLX antigens may provide the antigenic stimuli for persistent maternal humoral immunity.

Abortion, Habitual↗

Intergeneric Protoplast Fusion between Ruminococcus albus and an Anaerobic Recombinant, FE7.

Intergeneric protoplast fusion between Ruminococcus albus, a cellulolytic, gram-positive, anaerobic bacterium (Pc Sm Km), and an anaerobic recombinant, FE7 (Pc Sm Km), having lignin-related compound-degrading activities, was performed under strictly anaerobic conditions to introduce cellulase genes into strain FE7. The fusion frequency varied with different selected markers from 3.0 x 10 to 3.3 x 10. Two fusants, obtained from a synthetic medium with selective pressures of penicillin and streptomycin and with cellooli-gomer as the sole carbon source, were gram-negative rods. One of them, named FE7R2, showed 45 to 47% of the beta-glucosidase and cellobiosidase activities of its parent R. albus and still maintained a level of degradation activity against dehydrodivanillin, a lignin-related compound, of up to 87% of that of the parent strain FE7. To verify that the cellulolytic activities expressed in the fusant FE7R2 originated from R. albus cellulase genes, the beta-glucosidase gene of R. albus was cloned into Escherichia coli HB101 with plasmid pBR322. Cells bearing a recombinant plasmid, pRAII, produced high enzyme activities against both p-nitrophenyl-beta-d-glucoside and p-nitrophenyl-beta-d-cellobioside and could degrade cellobiose to glucose. Southern blot results showed that the cloned DNA fragment could hybridize with chromosomal DNAs of both R. albus and FE7R2, but did not with the chromosomal DNA of FE7, indicating that the beta-glucosidase gene fragment was introduced into the chromosome of FE7R2 from R. albus via the protoplast fusion. The fusant FE7R2 could utilize simultaneously both cellobiose and dehydrodivanillin. These results gave evidence that the fusion product FE7R2 is a recombinant strain between its parents R. albus and FE7. This recombinant has stably kept the above properties for about 2 years.

Journal Article↗

Cloning of the cellulase gene from Ruminococcus albus and its expression in Escherichia coli.

The gene for cellulase from Ruminococcus albus F-40 was cloned in Escherichia coli HB101 with pBR322. A 3.4-kilobase-pair HindIII fragment encoding cellulase hybridized with the chromosomal DNA of R. albus. The Ouchterlony double-fusion test gave a single precipitation line between the cloned enzyme and the cellulase from R. albus. The size of the cloned fragment was reduced by using HindIII and EcoRI. The resulting active fragment had a size of 1.9 kilobase pairs; and the restriction sites EcoRI, BamHI, PvuII, EcoRI, PvuII, and HindIII, in that order, were ligated into pUC19 at the EcoRI and HindIII sites (pURA1). Cellulase production by E. coli JM103(pURA1) in Luria-Bertani broth was remarkably enhanced, up to approximately 80 times, by controlling the pH at 6.5 and by reducing the concentration of NaCl in the broth to 80 mM.

Aerobiosis↗

Antigens of human trophoblast: trophoblast-lymphocyte cross-reactive antigens on platelets.

Human and rabbit antibodies to trophoblast-lymphocyte cross-reactive (TLX) antigens were employed in an enzyme-linked immunosorbent assay (ELISA) to identify and characterize the TLX alloantigen system on human platelets. Neither washing nor extraction in chaotrope or acid altered platelet TLX. The antigen was significantly changed by pronase and trypsin digestion, but Folch extraction yielded antigen in the hydrophilic interface, suggesting carbohydrate. Rabbit antibodies prepared to HLA-negative human syncytiotrophoblast TLX antigens were shown by platelet ELISA to have the same specificity and similar allotypy as anti-TLX antibodies from secondary (2 degrees) spontaneously aborting women. Patients with normal pregnancies before becoming 2 degrees aborters had both IgG and IgM antibodies to TLX. Anti-TLX in patients who never had a normal pregnancy were predominantly IgG. ELISA reactions performed with different concentrations of protein in the buffers detected anti-TLX activity in buffers containing high protein concentrations. This has been observed in studies of blocking antibodies in graft-versus-host disease and immune responses to tumor cells. Platelet TLX offers a new genetic and immunological approach to study similarities of the host-parasite relationships in pregnancy, transplantation, and cancer.

Abortion, Habitual↗

Studies on antilymphocyte antibodies in gestational choriocarcinoma.

The mixed lymphocyte reaction (MLR)-blocking activity of sera was examined in patients with gestational choriocarcinoma. The frequencies of human leukocyte antigens (HLAs) in seven choriocarcinoma patients and their husbands were not different from those in the normal Japanese population. In patients who were successfully treated, many mismatches were found between the woman's HLA-A, B, C antigens and those of her spouse. Sera from three patients showed cytotoxic activity against their husbands' T lymphocytes. Similar results were obtained with sera absorbed with platelets to test for cytotoxic activity against the husbands' B lymphocytes. Cytotoxic activity sometimes disappeared at the recovery stage of the disease. Significant MLR-blocking activity was found in sera of patients in a tumor-bearing stage, but was absent 3-4 months after serum human chorionic gonadotrophin levels dropped into the normal range. It reappeared when patients relapsed, showing that MLR-blocking activity reflected tumor burden. The MLR-blocking factors in patients' sera may not be identical to the so-called serum nonspecific immune inhibitor factors, but was specific to, or dependent on, the use of the husbands' lymphocytes as stimulator cells and was mediated by immunoglobulin (antibodies) reactive with antigens expressed on the husbands' B lymphocytes, e.g. the MHC class II antigens.

Antibodies↗

[Effect of oncofetal antigen-I (OFA-I) on lymphocyte response in pregnant women].

OFA-I, a membrane antigen on human malignant melanoma cells, was reported to be also found on a fetal brain tissue and to have a highly immunogenic effect on a pregnant woman. The aim in this study is to analyse an immunologic activity of OFA-I on in vitro response of lymphocytes from pregnant women. OFA-I was prepared from M-14 cells which are an in vitro cell line originating in a human malignant melanoma and purified by affinity column chromatography using the antibodies which were prepared from ascites of an embryonal carcinoma patient and exhibited anti-OFA-I activity in immune adherence test. The results obtained were as follows: No significant effect of OFA-I was found on PHA-induced lymphocyte response. A suppressive effect of OFA-I was observed on an unidirectional mixed lymphocyte culture (MLC) between a wife and husband when OFA-I was added at a concentration of more than 0.1 micrograms/ml. It was also suggested that the effect was attributable to inhibition in an initial phase of MLC. A suppressive effect of OFA-I was observed on Ig production of PWM-stimulated lymphocytes at a final concentration of more than 0.25 micrograms/ml. There was no significantly difference between the effect in pregnant and non-pregnant women. Furthermore, the effect was demonstrated to be mediated with macrophages in the culture system. From these findings, it was implied that OFA-I might play an important role in maternal immune recognition in the feto-placental unit.

Adult↗

[HLA antigens in choriocarcinoma].

Choriocarcinoma can be regarded as transplanted cancer, because its origin is in trophoblast which is fetal tissue. It is of interest whether HLA antigens are expressed on choriocarcinoma cells or not, since HLA antigens may play an important role if the patient's body could recognize choriocarcinoma as "not self" and initiate its immune response. In this report choriocarcinoma tissue in the uterus, obtained after hysterectomy, is stained by an indirect immunofluorescence technique using monoclonal antibodies to HLA-A,B,C and HLA-DR. We believe this is the first immunohistological study in which choriocarcinoma in utero is available for study and monoclonal antibodies to HLA antigens are applied. In the staining of HLA-A,B,C, and of HLA-DR, host cells, such as myometrial cells, show positive staining but choriocarcinoma cells show negative staining. It is thought that choriocarcinoma cells are viable and show negative staining of HLA antigens, on the basis of finding following HE staining and hCG staining using anti-hCG antibody and the clinical remarks of the patient such as her high urinary hCG titer before operation and no history of chemotherapy. These results agree with those of previous studies about HLA antigens on choriocarcinoma cell lines.

Animals↗