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Biomedical subjects

I Miyoshi

Publications and source records attributed to I Miyoshi.

At least 19 recordsLinked to original sources
PubMed1992

The establishment of an interleukin-6-dependent myeloma cell line (FLAM-76) carrying t(11;14)(q13;q32) chromosome abnormality from an aggressive nonsecretory plasma cell leukemia.

A new myeloma cell line designated FLAM-76 was established from a patient with an aggressive nonsecretory plasma cell leukemia. The cell line exhibited morphologic features of flaming cells and contained an abundant eosinophilic cytoplasm with many dilated cisternae of rough endoplasmic reticulum. FLAM-76 cells were positive for cytoplasmic kappa (kapp)-type immunoglobulin but did not secrete it into the culture medium. The cells proliferated in the presence of exogenous interleukin-6 (IL-6) and more than 800 pg/ml of IL-6 was necessary for their continuous growth. The cells did not grow without IL-6, and they did not produce IL-6. Thus, the growth of FLAM-76 appeared to be regulated by the paracrine mechanism of IL-6. Alpha-interferon (alpha-IFN) inhibited the IL-6-dependent growth of FLAM-76 in doses greater than 1000 U/ml. FLAM-76 cells expressed CD38 (OKT10) and cell adhesion-associated antigens such as CD44 and CD54 (ICAM-1). Chromosome analysis revealed FLAM-76 to have a hypodiploid chromosome constitution with t(11;14)(q13;q32) abnormality, which frequently is seen in neoplasms of B-cell origin. Immunoglobulin (JH and Ck) gene rearrangement (but no BCL-1 gene rearrangement) was found in this cell line.

Aged↗
PubMed1992

Inhibitory effects of human interferons on the immortalization of human, but not rabbit, T lymphocytes by human T-lymphotropic virus type-I (HTLV-I).

The effects of human interferon (IFN)-alpha, -beta, and -gamma on the immortalization of human and rabbit lymphocytes by human T-lymphotropic virus type-I (HTLV-I) have been investigated. The immortalization of human peripheral-blood lymphocytes co-cultured with lethally X-ray-irradiated HTLV-I-producer cells, MT-2, was blocked in the presence of more than 40 u/ml human recombinant IFN-alpha or more than 200 u/ml human natural type IFN-beta. However, rhIFN-gamma did not block immortalization by HTLV-I even at higher doses. On the other hand, the presence of high doses of hIFN-alpha, -beta, or -gamma did not exhibit any biological effect on the immortalization of rabbit peripheral-blood lymphocytes co-cultured with lethally X-ray-irradiated MT-2 cells. Integration of the full length of HTLV-I genome was detected in every transformant by Southern blot analysis. All cell lines established were CD4+/CD8 divided by T-lymphocytes, except for one cell line of CD4+/CD8+. Morphologically intact HTLV-I production was observed by electron microscopy in these cells. Our results indicate that HTLV-I released under the strongly suppressed condition in the presence of IFNs remains active and able to immortalize T lymphocytes. It is also suggested that immortalization of human T lymphocytes by HTLV-I can be inhibited by the antiviral state induced by the treatment with low doses of hIFN-alpha and -beta, whereas immortalization of rabbit T lymphocytes is not inhibited because of the species specificity of hIFNs.

Animals↗
PubMed1992

Establishment of a new Hodgkin's cell line (HD-70) of B-cell origin.

A new Hodgkin's cell line, designated HD-70, was established from the peripheral blood of a 69-year-old man with Hodgkin's disease of nodular sclerosing type. The cell line grows in a single cell suspension and has a doubling time of 28 hours. The cells have a round or irregular nucleus or multiple nuclei in relatively abundant cytoplasm that is positive for acid phosphatase, alpha-naphthyl butyrate esterase, and periodic acid-Schiff stains. HD-70 cells are positive for CD30 (Ki-1/Ber-H2), CD15 (Leu-M1), and CD71 (OKT9) antigens and contain cytoplasmic immunoglobulin (Ig) (A, kappa). Southern blot analysis showed that the cells have Ig heavy and kappa light chain gene rearrangement and lack T-cell receptor gene rearrangement. Chromosome analysis disclosed that the cells have a human karyotype with complicated abnormalities, including a 14q+. Heterotransplantation of the HD-70 cell line into newborn hamsters treated with antilymphocyte serum produced massive tumors with remarkable fibrosis and collagen band formation. These tumors displayed histologic features similar to those of the nodular sclerosing type tumor of the patient. Such fibrosis production and collagen band formation in heterotransplanted tumors suggest that a certain cytokine that induces fibrosis might be produced by HD-70 cells. This cell line may be useful for understanding the biology and pathogenesis of Hodgkin's disease.

Aged↗
PubMed1992

Establishment and characterization of a thymic carcinoma cell line (Ty-82) carrying t(15;19)(q15;p13) chromosome abnormality.

A new human cell line, designated Ty-82, was established from the pleural effusion of a 22-year-old woman with undifferentiated thymic carcinoma. This cell line consisted of primitive cells that were positive for alpha-naphthyl butyrate esterase and acid phosphatase. The cells were shown to express epithelial membrane antigen, but were completely negative for cytokeratin, carcinoembryonic antigen, glial fibrillary acidic protein, desmin, S-100 protein, lysozyme, Leu-7, HLA-DR (Ia), leukocyte common antigen, Ki-I antigen, T-cell antigens, B-cell antigens, myelomonocyte antigens, and Epstein-Barr-virus nuclear antigen. Electron microscopy showed that the cells were highly anaplastic, with no sign of cellular differentiation to any lineages. The Ty-82 cell line was found to have a karyotype of 46,XX,t(15;19)(q15;p13), being identical to that of the patient's tumor cells. Four of 5 nude mice inoculated sub-cutaneously with Ty-82 cells developed tumors which displayed a histological picture similar to the original tumor. Thymic carcinoma is a recently recognized entity, and its cellular and clinical behavior are poorly understood. The newly established thymic carcinoma cell line would provide a useful tool for the better understanding of this rare disease.

Adenocarcinoma↗
PubMed1992

IgD myeloma presenting as a testicular tumor: establishment and characterization of an IgD-secreting myeloma cell line.

A 72-year-old man presented with a left testicular tumor and underwent orchiectomy. The tumor was massively infiltrated with myeloma cells bearing monoclonal cytoplasmic IgD lambda. Three months after orchiectomy, he developed huge abdominal masses and subsequently ascites containing numerous myeloma cells. An IgD-secreting myeloma cell line, designated delta-47, was established from the ascites. This cell line expressed CD4 and CD38, but lacked Fc and complement receptors, surface immunoglobulin, CD19, HLA-DR, and PCA-1. CD30 was detected on the cultured cells but not on the ascites tumor cells. Delta-47 cells secreted the same immunoglobulin (IgD lambda) as was found in the patient's serum. The light chain had a molecular weight of 35 kD which was larger than that of the normal light chain. Chromosome analysis of delta-47 revealed an aneuploid karyotype with complex abnormalities including 1q+, 2p+, and 14q+. To our knowledge, this is the only IgD-secreting myeloma cell line and would provide a useful tool for the study of IgD production and IgD myeloma.

ADP-ribosyl Cyclase↗
PubMed1992

Segregation of human T cell lymphotropic virus type I and II infections by antibody reactivity to unique viral epitopes.

A recombinant protein of the human T cell lymphotropic virus type I (HTLV-I) gp46 outer membrane envelope, MTA-4 (residues 129-203), reacted by Western blot with sera from HTLV-I-infected individuals from the United States and Jamaica but not with 24 (10%) of 242 Japanese sera. A related gp46 recombinant protein, MTA-1 (residues 162-209), reacted with all 58 sera from HTLV-I-infected US and Jamaican individuals and 238 of 242 sera from infected Japanese (combined sensitivity of 99%). Neither recombinant showed reactivity to sera from HTLV-II-infected individuals or uninfected controls. The reactivity of recombinant proteins containing the region of HTLV-II gp46 analogous to MTA-1 was also evaluated by Western blot: GH2-K15 (residues 157-205) and GH2-K55 (residues 162-205) reacted with 88 (98%) and 89 (99%), respectively, of 90 sera from HTLV-II-infected individuals but not with sera from HTLV-I-infected individuals or uninfected controls. These recombinant proteins should permit the development of assays to unambiguously confirm and differentiate HTLV-I and HTLV-II infections.

Amino Acid Sequence↗
PubMed1992

Immunohistochemical detection of Pneumocystis carinii in transbronchial lung biopsy specimens: antigen difference between human and rat Pneumocystis carinii.

Transbronchial lung biopsy specimens, from three patients with non-AIDS-related Pneumocystis carinii pneumonia (PCP) and rat lung tissue in which PCP was induced by the administration of dexamethasone, were studied to determine the diagnostic usefulness of an immunohistochemical method using commercially available anti-Pneumocystis monoclonal antibody, 3F6, on formalin-fixed, paraffin-embedded tissue. PC was consistently stained a bright red color and unambiguously identified in all three human lung specimens, but not stained in lung tissues at autopsy from patients with various fungal pneumonias. In contrast, PC was weakly stained in PCP-induced rat lungs. The present study indicates human PC and rat PC to be antigenically different in terms of the existence of the 82 kilo-dalton (kD) antigen against which 3F6 is directed.

Adult↗
PubMed1992

Useful short-range transport of mouse embryos by means of a nonfreezing technique.

The survival of mouse morulae to be implanted on the uterine wall of a recipient after storage at 0 degree C in sucrose-containing medium has shown tendencies similar to those of blastocysts developing in vitro. The pregnancy rate, as defined by implantation sites per embryos transferred, has varied from 52.9 to 77.2% and decreased depending on the duration of storage (0 to 72 h), and was lower than the rate of blastocyst formation in vitro (57.2 to 87.3%). Regarding the birth rate, 56.0 to 59.4% of implanted embryos were delivered as live pups regardless of storage duration, with the exception of controls (78.9%, 0 h). Eventually, 37.0, 32.8, or 29.9% of embryos were obtained as live pups when preserved by our system for 24, 48, or 72 hours, respectively. This viable, highly reproducible nonfreezing technique is useful for embryo preservation. Embryos were transported to evaluate the efficacy of the nonfreezing method. Mouse morulae were kept in a thermos bottle filled with ice-water and packaged in polystyrene foam. The package was transported from Kochi to Sapporo, about 1300 km, within 48 hours during which it was possible to maintain 0 degree C in the bottle and deliver to most areas in Japan. After shipping, the survival of embryos was 46.8% for development to blastocysts in vitro, and 37.1% for the pregnancy rate, with a final birth rate of 14.1%. Transport factors other than storage may have contributed to the lower birth rate; however, our system is cheaper, more convenient, and more practical than others for short-range transport.

Animals↗
PubMed1992

Unidirectional distribution of mosaicism in chimeric rats.

Experimental rat chimeras were produced by aggregation of eight-cell embryos from two inbred strains, ACI/Hkm and WKAH/Hkm, which differ from each other in their major histocompatibility complexes and coat colors, and their mosaicism was analyzed. The existence of the isozyme Es-1, a serum cholinesterase specifically produced by WKAH-derived cells, and the agouti coat color due to ACI cells, indicated that all of the rats analyzed were unequivocal chimeras. The proportion of ACI cells in the red blood cell populations of the chimeras varied from 45% to 98%, as determined with a fluorescence-activated cell sorter and a monoclonal antibody against class I (RT1) antigen. Digital analysis of the coat color revealed that the proportion of the ACI type of coat color ranged from 72% to 98% in these chimeric rats. Each phenotype expressed in the coat color was complex and varied in size. The ratios of red blood cells and the coat color inclined toward the ACI type of cell population. Conversely, the rate of the WKAH-cell-type population was less than 50%. A breeding test disclosed chimerism of germ cells in two chimeric rats, and there were more pups with agouti coats than with albino coats. Taken together, it was shown in most of the phenotypes analyzed that the ACI type of cells was predominant in all of the chimeric rats. We discuss the possible causes for this unbalanced distribution in the rats.

Animals↗
PubMed1992

Immunoglobulin prophylaxis against HTLV-I in a rabbit model.

We have investigated the protective effect of human T-cell leukemia virus I (HTLV-I) immune globulin (HTLVIG) against HTLV-I in rabbits. HTLVIG containing 77 mg/ml of IgG was prepared from pooled plasma from seropositive healthy persons. In the first experiment, four groups (A, B, C, and D) of three rabbits were transfused with 5 ml blood from an HTLV-I-infected rabbit. Groups A, B, and C were infused 24 h later with 10, 5, and 2 ml HTLVIG, respectively, while group D was infused with 10 ml HTLVIG 48 h later. Seroconversion for HTLV-I occurred in none of group A, one of group B, and all of groups C and D after 2-5 weeks. In the second experiment, four litters (E, F, G, and H) born to another virus-infected rabbit and consisting of 7, 5, 7, and 7 newborns, respectively, were used. Litters E and H were allowed to grow normally as controls, while litters F and G were given intraperitoneal inoculation of 3 ml/kg of HTLVIG weekly four times until weaning. Although three of litters E and H each seroconverted after 5-8 weeks, none of litters F, and one of litter G became antibody-positive after 10 weeks. Presence or absence of HTLV-I infection in all these animals was confirmed by transfusion assay or gene amplification. These results indicate that passive immunization protects rabbits against blood- and milk-borne transmission of HTLV-I.

Animals↗
PubMed1992

Effects of sex hormones on fulminant hepatitis in LEC rats: a model of Wilson's disease.

LEC rats, which have hereditary hepatitis and have recently been proposed as an animal model for Wilson's disease, were examined to determine the effects of sex hormones on fulminant hepatitis. After the rats had undergone ovariectomies or orchidectomies (castration) and were compared with intact rats, the age at the onset of fulminant hepatitis was not substantially altered but the survival rates decreased from 50% to 12.5% for females and 75% to 14.3% for males, indicating that sex hormones did not influence the occurrence of fulminant hepatitis but influenced mortality due to fulminant hepatitis. When testosterone was administered to the ovariectomized or orchidectomized rats, the survival rate increased to over 90% in both sexes. In contrast, estradiol did not affect the survival rate of either sex but affected the onset of fulminant hepatitis. That is, with the administration of estradiol, the age at which serum GPT activity reached its maximum was delayed 4 weeks in ovariectomized rats and 6 weeks in orchidectomized rats as compared with intact rats. A similar but somewhat weaker tendency appeared in rats given progesterone. The results of our study indicate that sex hormones have no effect on the rate of occurrence of hepatitis but affect the progression of hepatitis. In particular, testosterone increased the survival rate of rats with fulminant hepatitis, and exogenous estradiol delayed the onset of hepatitis for several weeks.

Alanine Transaminase↗
PubMed1992

Gene rearrangement and overexpression of PRAD1 in lymphoid malignancy with t(11;14)(q13;q32) translocation.

The proto-oncogene PRAD1 (parathyroid adenoma 1) on chromosome 11q13 was found to be overexpressed in all five B-cell lines with t(11;14)(q13;q32) translocation tested. One B-cell lymphoma and four myeloma cell lines with this translocation demonstrated more than 10-fold overexpression as determined by Northern blot analysis, when compared with normal lymphoid tissues such as thymus, spleen and lymph node. Hematopoietic cell lines without the translocation were also examined, but none of these demonstrated the overexpression, confirming that overexpression of the PRAD1 gene is associated with t(11;14) translocation. A truncated form of mRNA was seen in one of five cell lines with the translocation, SP-49. Hybridization with different regions of the PRAD1 cDNA revealed that the truncated form of mRNA retained the coding region but had lost the 3' untranslated region. Southern blot analysis demonstrated a gene rearrangement in this SP-49 cell line. To study the genetic alteration responsible for the truncated form of mRNA in this cell line, the rearranged allele as well as the germline allele were cloned. The restriction map revealed that the rearranged portion was at the 3' end of the PRAD1 gene, eliminating the mRNA-destabilizing signal AUUUA. Human-rodent hybrid cell analysis demonstrated that the region introduced 3' of PRAD1 was derived from chromosome 11, suggesting that the PRAD1 gene region is deleted at the 3' end. Over-expression of the PRAD1 gene in association with t(11;14)(q13;q32) translocation suggested that in these cases the regulation of PRAD1 was altered by the juxtaposed gene, most likely the immunoglobulin heavy-chain gene from chromosome 14.

Base Sequence↗
PubMed1992

Acute megakaryoblastic leukemia: establishment of a new cell line (MKPL-1) in vitro and in vivo.

A megakaryoblastic cell line (MKPL-1) was newly established from the bone marrow of an adult patient with acute megakaryoblastic leukemia. This cell line grew in single cell suspension with a doubling time of 30 h and consisted of large primitive blasts with persistent development of giant cells carrying multilobed nuclei. MKPL-1 cells were positive for platelet GPIIb/IIIa (CD41) and GPIIIa (CD61), and expressed OKM5 (CD36), MY7 (CD13), and MY9 (CD33) antigens in the absence of erythroid and lymphoid markers. The cytochemical and morphologic characteristics of MKPL-1 were also consistent with those of megakaryoblasts. The cells did not, however, express ultrastructural platelet peroxidase which is considered to be another marker of the megakaryocytic lineage. Cytogenetic analysis of MKPL-1 revealed a model chromosome number of 92 with abnormal chromosomes including those found in the patient's bone marrow cells. Furthermore, MKPL-1 cells were serially transplanted into nude mice for nine passages with production of lethal tumors and leukemic manifestation. Thus, our megakaryoblastic cell line which can be maintained both in vitro and in vivo would be useful for further studies of the biology of megakaryopoiesis and megakaryoblastic leukemia.

Aged↗