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Biomedical subjects

R Ueda

Publications and source records attributed to R Ueda.

At least 361 records · Page 20Linked to original sources
PubMed1985

Preliminary clinical trial of autologous bone marrow transplantation after in vitro monoclonal antibody and complement treatments in null cell-type acute lymphocytic leukemia.

Autologous bone marrow transplantation (BMT) in null cell-type acute lymphocytic leukemia (Null-ALL) was carried out after depletion of leukemia cells from transplanted bone marrow. Patients' autologous bone marrow cells were harvested during remission and treated in vitro with complement and three monoclonal antibodies (NL-1, NL-22 and HL-47) reactive to Null-ALL cells, and then cryopreserved. Three patients were transplanted with the antibody-treated bone marrow cells during the first remission period after preconditioning with intensive chemotherapy and total body irradiation, while transplantations in two other patients, who were in poor clinical condition, were done during the fourth remission period and the third relapse, respectively. Good preservation of hematopoietic stem cells after antibody treatment and cryopreservation of bone marrow cells was demonstrated in all five cases studied. Clinically, prompt recovery of white blood cells and platelets was observed in the three patients who received BMT during the first remission period; two of them have continued remission (2 and 15 months), while the other relapsed after 7 months of remission. these results suggested that autologous BMT with these three antibodies may be an effective mode of therapy for Null-ALL patients.

Antibodies, Monoclonal↗
PubMed1984

Human glioma associated and related antigens, identified by monoclonal antibodies.

Tumour specific and various tumour-associated antigens expressed on human glioma cells were reported from many laboratories. These have been identified by xeno-, allo-, or auto-sera and more recently by utilizing monoclonal antibodies produced by hybridoma technique. In this article, the results of reports are summarized comparing the studies with conventional antisera and monoclonal antibodies. The present and future clinical applications of these monoclonal antibodies are described.

Antibodies, Monoclonal↗
PubMed1984

Nuclear immunofluorescence by a monoclonal antibody against microtubule-associated protein-1 as it is associated with cell proliferation and transformation.

Monoclonal antibody against microtubule-associated protein-1 produced intranuclear immunofluorescent spots, which disappeared under growth-inhibited conditions caused by serum starvation and saturated cell density in untransformed cells. A change of medium to 10% serum gave rise to the reappearance of nuclear spots before the resumption of DNA synthesis. This reversible change of immunofluorescence was also caused by a temperature shift in rat 3Y1 cells transformed by Simian virus-40-A640 (temperature-sensitive in large T-antigen). The fluorescence decreased during S phase of the cell cycle. In contrast the transformed cells always showed nuclear fluorescence, irrespective of serum concentrations or the cell cycle. Growth-inhibited cells previously treated with detergent and salt revealed nuclear fluorescent spots. This result suggested antigenic modification.

Animals↗
PubMed1984

Five antigens on human T cells detected by mouse monoclonal antibodies.

Five antigen systems were defined by the monoclonal antibodies (MoAb) produced against mature T cells. The antigens recognized were grouped into two categories based on the antigen distribution on T cells. (a) Tp 120 [mol. wt 120 kilodaltons (120kD)] and Tp40 (40kD), these are on most peripheral T cells, but not on any other cell lineages, i.e. pan-T antigen. (b) Ts32 (32kD), Ts145 (145kD) and TsA (not determined), these antigens are present only on certain populations of peripheral T cells, i.e., T subset antigen. Among these five, Ts145 and TsA are probably novel T cell antigens. Cell surface phenotypes of leukaemias and lymphomas were typed with these MoAb. Ia like antigen negative, null cell type acute lymphocytic leukaemia (Ia- null ALL) are Tp40+, suggesting that this type of ALL belongs to a T cell lineage. T cell ALL (T-ALL) and lymphoblastic lymphoma (LL) were both Tp40+, Ts32+, TsA+ and a half of the cases were Tp120+, but the expression of Tp40 was stronger on LL cells. Mature T cell (T2) lymphoma and adult T cell leukaemia (ATL) were Tp120+, TsA+, while Tp40 was weakly expressed on only one third of the cases. These MoAb were found to be useful to estimate the origin of various T cell malignancies.

Animals↗
PubMed1983

Polymorphism and stability of histone gene clusters in Drosophila melanogaster cultured cells.

In a previous communication (Saigo, K., Millstein, L. and Thomas, C.A., Jr. (1981) Cold Spring Harbor Symp. Quant. Biol. 45, 815-827), the overall structure of histone genes of Schneider line 2 cells was shown to extensively differ from that of Oregon-R embryo from which the cell line was established, and it was speculated that the histone genes might be reshuffled extensively during either the periods of the establishment, or maintenance of cell lines, or both. To establish the validity of this notion the structure of histone genes was examined in Drosophila melanogaster cultured cells. The overall organization of histone gene clusters was found to be stably maintained in both the periods for the establishment and maintenance of cultured cells, indicating that the previous assumption is inadequate. Instead of an extensive rearrangement, minor structural changes were found to occasionally occur probably by simple base substitutions and/or, deletion or insertion of very short DNA pieces. It was also shown that the extensive variation in structures of histone genes in cultured cells such as Schneider line 2 are attributable to polymorphism on the level of individual flies.

Animals↗
PubMed1983

Serological analysis of cell surface antigens of HL-60 cells before and after treatment with a phorbol ester tumor promoter.

The human HL-60 cell line derived from acute promyelocytic leukemia, consisting of promyelocytic type of cells, was able to differentiate into adherent cells with monocytemacrophage features by the treatment with 12-0-tetradecanoyl phorbol-13-acetate (TPA). Cell surface antigens of HL-60 cells before and after TPA treatment were studied with monoclonal antibodies and four hybridoma clones producing IgM antibodies were established. Two antibodies (HL-21 and HL-47) reacted only with the immunizing TPA-treated HL-60 cells, and HL-1 antibody produced against untreated cells was reactive with both TPA-treated and untreated cells, but HL-5 antibody reacted predominantly with the immunizing untreated cells. Serological reactivity against various types of normal hematopoietic cells and acute leukemias (diagnosed by the French-American-British classification) was studied by immune adherence assay and immuno-electron microscopy. HL-21 antibody was reactive with monocytes and most cases of M4 and M5 types of acute non-lymphocytic leukemia cells. HL-47 antibody did not react with the cells of myelocyte-monocyte lineage or mature lymphocytes, but it did react with one-third of acute lymphocytic leukemia (L1 and L2) cases. Since all HL-47+ cases were included in the group of common ALL antigen positive cases, it was estimated that HL-47 is a differentiation antigen present on lymphocyte precursors, from which null-cell type acute lymphocytic leukemia cells generally originate. HL-1 antibody reacted with the cells of myelocyte-monocyte lineage as well as those of most acute non-lymphocytic leukemias. HL-5 antibody reacted with granulocytes and M2 type of acute myelocytic leukemia cases, and also with M5 type of acute monocytic leukemia cases. Serological studies of these antibodies revealed that TPA can induce to differentiate HL-60 cells not only into HL-21+ macrophage-like cells, but also into HL-47+ lymphoid stem cells. In addition, these antibodies were demonstrated to be very valuable for differential diagnosis of acute leukemias.

Antibodies, Monoclonal↗
PubMed1983

Two monoclonal antibodies detecting allotypic determinants of HLA-A.

Three mouse hybridomas producing cytotoxic antibodies against HLA were established. By standard microcytotoxicity test against panels of normal controls, the antigen defined by MA-9 antibody (IgM) showed a good correlation with HLA-A9 alloantigen detected by conventional typing alloantisera (r = 1.0). Family studies also showed that MA-9 determinant segregated with HLA-A9. MA-10 antibody (IgM) reacted with all HLA-A10 positive lymphocyte donors and cross-reacted with two thirds of HLA-AW33 positive donors. Ml-1 antibody (IgG2a) reacted with all the panel cells tested and immunoprecipitated a molecule of 43,000 daltons from Nonidet P-40 lysates of 3H-glucosamine-labelled cells. The results showed that MA-9 and MA-10 antibodies can be used as routine tissue typing reagents.

Adult↗
PubMed1983

Monoclonal antibody against microtubule associated protein-1 produces immunofluorescent spots in the nucleus and centrosome of cultured mammalian cells.

A monoclonal antibody was raised against the highest molecular weight protein associated with microtubules (MAP-1). Its specific binding to MAP-1 was determined by immunoblotting of the gel electrophoretogram of microtubule proteins prepared from porcine brain. The antibody reacted only with MAP-1, not with MAP-2, tau or tubulin. Indirect immunofluorescent staining by this antibody showed bright intranuclear spots, the centrosome and the faint meshwork of the cytoplasm in several types of cultured mammalian cells; HeLa, PtK2, human skin fibroblasts, mouse melanoma cells, Chinese hamster ovary cells. The nuclear spots in the interphase cells, were replaced by diffuse enhanced fluorescence throughout the cell except for chromosomes during mitosis. They reappeared in late telophase, first in the cytoplasm, late in the nucleus. The punctate pattern of nuclear immunofluorescence was not affected by microtubule-depolymerizing agents. The result that it persisted on residual cell structures after extraction with a high salt concentration buffer containing Triton X-100 followed by digestion with DNase I and RNase A suggests that the antigen is associated with the nuclear skeleton.

Animals↗
PubMed1983

[Analysis of cell surface antigen of kidney cancer by autologous sera and monoclonal antibodies].

Sera from renal cancer patients were tested for reactivity with surface antigens of cultured autologous renal cancer cells, so called "autologous typing". Reactive sera from six patients were analyzed by absorption tests with autologous, allogeneic, and xenogeneic normal and malignant cells. The absorption analysis indicated the detection of three classes of surface antigens by autologous sera, i.e. class 1, individually distinct tumor antigens, class 2, shared tumor antigens, and class 3, normal cell surface antigens. Seventeen monoclonal antibodies derived from fusions with spleen cells of mice immunized with renal cancer cell lines, which showed the class 1 or class 2 antigens, identified nine cell surface antigenic systems. None of the mouse antibodies identified in this study was succeeded in recognizing antigens that are tumor specific. Three of antigenic systems, however, showed the specificity for renal tissue, and seemed to be useful for immunopathological diagnosis.

Animals↗
PubMed1983

[Subclassification of acute leukemia and monoclonal antibodies].

Recent advances in immunological techniques, particularly, the development of monoclonal antibodies by the hybridoma technique, have contributed a great to the understanding of cell surface antigens of human hematopoietic cells. We have produced monoclonal antibodies detecting the following antigens: Ia-like (NL-12, MD-1A), Null-ALL associated (NL-1, NL-22, HL-47), T cell (ATL-27, Thy-64), myelocyte-monocyte (HL-1), and monocyte (HL-21). Using these antibodies, serological typing of leukemia was performed and it was found to facilitate considerably the diagnosis of leukemia. In consideration of therapeutic use of two antibodies detecting Null-ALL associated antigen (NL-1, NL-22), reactivity against major organs was studied by immunoperoxidase staining. NL-1 antibody was reactive with glomeli and tubules of kidney and epithelial cells of small intestine, while NL-22 antibody reacted with tubules of kidney, liver cells and some other tissues. This results suggested the difficulty of application of these antibodies directly to the patients. Reactivity against colony forming cell in bone marrow was also examined and moderate inhibition was observed by both antibodies, which might allow to eliminate ALL cells in cryopreserved autologous bone marrow cells from bone marrow transplantation patients by the treatment with appropriate concentration of antibodies.

Acute Disease↗
PubMed1982

Serological analysis of cell surface antigens of null cell acute lymphocytic leukemia by mouse monoclonal antibodies.

Nine antigens systems were defined. Two were related to HLA-A,B,C and to Ia-like antigens; the others could be grouped into three categories. (i) NL-22, NL-1: NL-22 antibody reacted with leukemia cells from 12 to 16 cases of null cell acute lymphocytic leukemia (null-ALL) but not with any other type of leukemia tested or with lymphoid cells of various origins. Among cultured cell lines tested, one (NALM-6) of three null-ALL cell lines was positive, the others were negative. Absorption analysis confirmed the restriction of NL-22 antigen to null-ALL. NL-1 antibody was reactive with leukemia cells from 10 to 16 cases of null-ALL and 3 of 6 cases of chronic myelocytic leukemia in blastic crisis (CML-BC). The antigen was present also on a minor population of normal lymphoid cells. The distribution and molecular weight (100,000; glycoprotein) of the NL-1 antigen resembled that of the previously described common ALL antigen (cALL). (ii) NL-30, NL-4: Both antibodies exhibited almost identical patterns of reactivities against cultured cell lines tested. They reacted with leukemia cells from some cases of null-ALL, adult T-cell leukemia, and CML-BC, although they showed discordance in their reactivities against a panel of leukemia cells, (iii) NL-9, NL-8, NL-25: These three antibodies detect serologically distinguishable determinants on a broad range of leukemias and normal lymphoid and hematopoietic cell types. The antibodies analyzed in this study provide evidence for the heterogeneity of null-ALL by demonstrating a variety of antigen phenotypes on leukemia cells. One of the antigens (NL-22) appears to be restricted to null-ALL.

Animals↗
PubMed1982

Induction of pole cells in sterilized Drosophila embryos by injection of subcellular fraction from eggs.

A subcellular fraction isolated from a homogenate of young Drosophila embryos was shown to be capable of inducing pole cells when injected into UV-sterilized Drosophila embryos. Most of the pole cell-inducing activity was recovered from the precipitate after centrifugation at 27,000 x g. The activity remained in this precipitate (called F-3 fraction hereafter) even after membranous structures were removed from it through centrifugation on a sucrose density gradient. Dialysis, lyophilization, and heating at 80 degrees C for 10 min did not inactivate the F-3 fraction. The pole cells, which were produced when the F-3 fraction was injected at the posterior pole of UV-sterilized embryos, did not develop into germ cells. Furthermore, the F-3 fraction was unable to induce pole cells when injected into the anterior region of the egg. These results can be explained by assuming that (i) pole cell formation and germ cell determination are controlled by different factors, (ii) pole cell formation requires at least two factors, which are normally localized in the posterior-pole cytoplasm, one of which is sensitive and one resistant to the UV dosage we used, and (iii) the subcellular fraction we obtained contains the UV-sensitive factor but not the UV-resistant factor.

Journal Article↗
PubMed1982

[Flow cytometric analysis of cell surface antigens on leukemia cells by monoclonal antibodies and its application].

Nine monoclonal antibodies against null cell ALL cells were obtained by mouse hybridoma technique. Using two antibodies, namely B-1 and D-22, out of these, cell surface antigens on various leukemia cells, normal bone marrow cells, spleen cells and cultured cells were analysed by means of flow cytometry. B-1 antibody was found to have reactivities against null cell ALL cells as well as some CML-blastic crisis cells. The antibody reacted against some cultured lymphoid cell lines and also several percent of bone marrow cells. The reactivities of B-1 antibody were quite similar to those of c-ALL by Greaves et al and J-5 by Ritz et al. Radioimmunoprecipitation experiment suggested that B-1 antibody define the same antigen as the other two do. Meanwhile, D-22 antibody reacted against null cell ALL cells, but did not react against any other types of leukemia cells or any other cultured cell lines, or bone marrow cells. Thus D-22 antibody was considered to have higher specificity for null cell ALL cells than any other antibodies reported. On the other hand, the analyses by flow cytometry were well correlated with the analysis by immune adherence assay and by immunofluorescent microscopy. Besides, the analysis by flow cytometry provided us precise quantitative data and histogram pattern of positive cells. Furthermore, detailed examination of subpopulation could be done using cell sorter or other biological features could be analysed simultaneously using another fluorescent dye. Although, it has a few limitations, flow cytometry could take over other methods which are currently used in this field. Finally, the possibility of this method for the clinical diagnosis of leukemia was discussed.

Antibodies, Monoclonal↗
PubMed1981

Comparison of [3H]glucosamine-labeled glycoproteins from human renal cancer and normal kidney epithelial cell cultures by two-dimensional polyacrylamide gel electrophoresis.

Two-dimensional polyacrylamide gel electrophoresis has been used to compare the Nonidet P-40 soluble, [3H]glucosamine-labeled glycoproteins of human kidney cancer cell lines and short-term cultures of normal kidney epithelia. Two pairs of autologous combinations of tumor and normal cells and also six other cancer lines and four normal cultures were examined. Because autologous cells could be compared, possible differences due to allogeneic variations could be eliminated. The electrophoretic patterns given by tumor and normal cells were extremely similar. Nevertheless, more than 10 differences could be recognized when randomly selected combinations of cancer and normal cells were examined in detail. Of these differences, however, only one was consistently found in all possible combinations of cancer and normal cells. Cancer cells had three prominent components of molecular weight 27,500 and pIs of 5.7, 5.3, and 4.9, respectively. Normal cells have a fainter and different constellation of spots in this region, with the more acid component (pI 4.9) being absent (or barely detectable). The pattern of fetal kidney cells resembled that given by normal adult kidney.

Cell Line↗