研究者詳細 - 増子　正義

Expression of coxsackievirus and adenovirus receptor in hearts of rats with experimental autoimmune myocarditis 査読

M Ito, M Kodama, M Masuko, M Yamaura, K Fuse, Y Uesugi, S Hirono, Y Okura, K Kato, Y Hotta, T Honda, R Kuwano, Y Aizawa

CIRCULATION RESEARCH 86 ( 3 ) 275 - 280 2000年2月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：LIPPINCOTT WILLIAMS & WILKINS

The expression of coxsackievirus and adenovirus receptor (CAR) was dominant in the brains and hearts of mice until the newborn phase. There is no detailed information concerning the relation between the expression of CAR and development of hearts. It is also uncertain whether CAR is able to be induced in adult hearts after cardiac injury. We demonstrated that CAR was abundant in the hearts of newborn rats but was barely detectable in the hearts of adult rats. The expression of CAR in rat hearts with experimental autoimmune myocarditis, which was induced by immunization of purified cardiac myosin, was serially investigated. Active myocarditis was observed from day 15 after immunization. By immunohistochemistry, cardiomyocytes were strongly stained for CAR antibody from days 24 to 42. CAR mRNA was also detected from days LX to 30 by using reverse transcription-polymerase chain reaction. In the next experiment, the induction of CAR on isolated cardiomyocytes was investigated. CAR was barely detectable in cultured cardiomyocytes by Western blot analysis after isolation. This molecule gradually appeared along with the creation of clusters and beating of cardiomyocytes. Furthermore, the induction of CAR in cultured cardiomyocytes increased after supplement with conditioned medium of rat splenocytes activated by concanavalin A. In conclusion, rat CAR is expressed strongly in the hearts of newborn rats and is suppressed in those of adult rats. The expression of CAR is enhanced during the active phase of experimental autoimmune myocarditis and is induced by inflammatory mediators, CAR may play a role in cell-to-cell contact and adhesion of cardiomyocytes.

Web of Science

Cyclosporine A-associated fatal central nervous system angiopathy in a bone marrow transplant recipient: An autopsy case 査読

T. Koide, M. Yamada, T. Takahashi, S. Igarashi, M. Masuko, T. Furukawa, T. Kuroha, T. Koike, M. Sato, R. Tanaka, S. Tsuji, H. Takahashi

Acta Neuropathologica 99 ( 6 ) 680 - 684 2000年

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Springer Verlag

We report here the case of a 32-year-old woman who suffered from a unique angiopathy in the central nervous system (CNS). She died of multiple infarcts in the brain stem and cerebellum during treatment with cyclosporine A after bone marrow transplantation for refractory anemia with excess of blasts. The autopsy findings showed segmental narrowing of the basilar artery, in which circumferential dissection of the internal elastic lamina had occurred. The distal portion of the basilar artery was obstructed by upward dislocation of the dissected intima. Similar angiopathy was also observed at multiple sites along the basilar artery branches. These findings suggest endothelial damage, including vasoconstriction and dissection of the CNS arteries.

DOI： 10.1007/s004010051179

DOI： 10.1002/(SICI)1097-0320(19990301)35:3<249::AID-CYTO8>3.0.CO;2-O

Novel technique for the direct flow cytofluorometric analysis of human basophils in unseparated blood and bone marrow, and the characterization of phenotype and peroxidase of human basophils 査読

Ken Toba, Tadashi Koike, Akira Shibata, Shigeo Hashimoto, Masuhiro Takahashi, Masayoshi Masuko, Takaaki Azegami, Hidenobu Takahashi, Yoshifusa Aizawa

Cytometry 35 ( 3 ) 249 - 259 1999年3月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Wiley-Liss Inc.

Background: No technique has been reported to analyze directly the antigen expression on basophil leukocytes when using a flow cytometer
therefore, the exact phenotype of human basophils and the character of the peroxidase in basophils are not well understood. Methods: Human blood basophils were purified by using an antibody against high-affinity Fc epsilon receptor (hFcεR) and a MACS(TM) magnetic cell sorting system and then cytochemically stained. The phenotype and peroxidase of the human basophils were flow cytofluorometrically analyzed directly in unseparated blood and bone marrow samples as hFcεR+/MBP+ (major basic protein)/Hist+ (histamine) light-density cells distributed in the high sidescatter area of lymphocytes on light scattergrams. Results: The peroxidase granules of human basophils were stained by an anti-eosinophil peroxidase (EPO) antobody. The human blood basophils had common granulocyte markers plus CD25, i.e., they were CD11a/CD1lb/CD11c/CD25/CD38/CD13/CD33/hFcεR/MBP/Hist/EPO positive, CD71 dim positive, CD14/CD15 partially positive, and CD2/CD3/CD7/CD122/CD16/CD56/CD57/CD10/CD19/CD20/CD22/HLA-DR/MPO (myeloperoxidase)/CD23 negative. Further examination was done to analyze the expression of colony-stimulating factor receptors on three lineages of granulocytes, i.e., basophils, eosinophils, and neutrophils. The neutrophils were CD114 (G-CSFR)/CD116 (GM-CSFR)/CD124 [interleukin (IL)-4R]/CD126 (IL- 6R) positive and CD123 (IL-3R)/CD125 (IL-5R) negative. In contrast, the eosinophils and basophils were CD116/CD123/CD125/CD126 positive and CD114/CD124 negative. Conclusions: This novel technique for directly characterizing human basophil leukocytes with flow cytometry may be a convenient way to screen the expression of surface antigens and the cytoplasmic expression of CD antigens and other proteins in human blood basophils and to analyze alterations of the character of basophils by cytokines and other biological substances in vivo and in vitro.

Expression of eosinophil peroxidase in the immature basophil cell line KU812-F 査読

M Masuko, T Koike, K Toba, K Kishi, T Kuroha, T Furukawa, Fuse, I, M Takahashi, R Kuwano, A Shibata, Y Aizawa

LEUKEMIA RESEARCH 23 ( 2 ) 99 - 104 1999年2月

担当区分：筆頭著者,　責任著者記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：PERGAMON-ELSEVIER SCIENCE LTD

Although peroxidase activity in basophils can be detected by optical and ultrastructural cytochemistry, its characteristics remain to be determined. We have demonstrated the characteristics of peroxidase activity induced in the immature basophil cell line, KU812-F. Ultrastructurally, peroxidase activity was detected in granules as well as in the perinuclear space and endoplasmic reticulum. Immunocytochemistry revealed that KU812-F cells were stained by anti-eosinophil peroxidase antibodies, and eosinophil peroxidase mRNA, not myeloperoxidase, was detected in the cells using Northern hybridization and reverse transcription-polymerase chain reaction. Eosinophil peroxidase can be one of the molecules shared with eosinophils and basophils. The biological function of eosinophil peroxidase detected in basophils remains uncertain. (C) 1999 Elsevier Science Ltd. All rights reserved.

Web of Science

Hematopoietic cytokine-dependent differentiation to eosinophils and neutrophils in a newly established acute promyelocytic leukemia cell line with t(15;17) 査読

K. Kishi, K. Toba, T. A. Azegami, N. Tsukada, Y. Uesugi, M. Masuko, H. Niwano, S. Hashimoto, M. Sakaue, T. Furukawa, T. Koike, H. Takahashi, T. Maekawa, T. Abe, Y. Aizawa

Experimental Hematology 26 ( 2 ) 135 - 142 1998年

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Elsevier Inc.

We recently established an acute promyelocytic leukemia (APL) cell line (HT93) that has the capacity to differentiate into neutrophils and eosinophils in response to all-trans retinoic acid (ATRA) and human hematopoietic cytokines. The cells had a myeloblastic morphology, were positive for surface CD33, CD34, and CD56, and showed the following karyotypes: 46, XY, t(1
12)(q25
p13), 2q+, t(4
6)(q12
q13), and t(15
17)(q22
q11). When the cells were cultured with ATRA, they showed nuclear segmentation and developed secondary granules consisting in part of neutrophils and eosinophils. In the presence of ATRA and granulocyte colony- stimulating factor (G-CSF), the cells showed polymorphonuclear neutrophil differentiation accompanied by expression of surface CD11b, CD15, CD10, positive activity for neutrophil alkaline phosphatase (NAP), anti NAP mRNA expression. In cultures with ATRA and granulocyte-macrophage colony- stimulating factor (GM-CSF), IL (interleukin)-3, or IL-5, HT93 showed remarkable eosinophil maturation at day 8 as determined by luxol fast blue staining, in addition to expression of eosinophil peroxidase and major basic protein. These results indicate that HT93 is an APL cell line with the ability to differentiate into neutrophils anti eosinophils, and that these lineages are dependent on the CSF added. HT 93 should prove to be a useful model in analyzing the effects of hematopoietic cytokines on proliferation, differentiation, and maturation of hematopoietic progenitors.

Confiscation of autonomous proliferation from p2l0bcr-abl transfected cells by re-transfection with DNA for homologous recombination

M. Takahashi, N. Shigeno, N. Suzuki, H. Takahashi, K. Nikkuni, M. Masuko, M. Sakaue, T. Furukawa, K. Kishi, T. Koike, Y. Aizawa

Transgenics 2 ( 3 ) 241 - 247 1998年

記述言語：英語掲載種別：研究論文（学術雑誌）

Ideally, somatic gene therapy for genetic diseases is designed to replace an abnormal gene with a normal gene and to normalize the characteristics of abnormal cells. Up to now, repair of an abnormal gene can be accomplished only by homologous recombination between that abnormal gene and the gene for correction. Our study was designed to discover whether or not homologous recombination occurs in hematopoietic cells and whether it is possible to establish gene therapy using homologous recombination. A murine hematopoietic cell line, FDC-P2, which was factor-dependent but became factor-independent by being transfected with p210bcr-abl was used for the target cell for homologous recombination. The plasmid for homologous recombination was made by inserting the neo-gene in the region of bcr-abl, thymidine kinase-gene in the region outside of bcr-abl, and placing the bcr-abl region reversely for defining the homologous recombination region to bcr-abl. Transfection was done by electroporation and selection was performed by adding G418 and gancyclovir. One clone was established and confirmed to be replaced by the plasmid made for homologous recombination by RT-PCR findings of thymidine kinase (-), neo (+), bcr-abl (-) and a reasonable change of the band by Southern blot analysis. The clone became factor-independent. These findings suggest the possibility of homologous recombination-associated gene replacement therapy for hematological diseases including leukemia.

Primary familial polycythaemia associated with a novel point mutation in the erythropoietin receptor 査読

Tatsuo Furukawa, Miwako Narita, Minori Sakaue, Tomio Otsuka, Takashi Kuroha, Masayoshi Masuko, Takuaki Azegami, Kenji Kishi, Masuhiro Takahashi, Jirou Utsumi, Tadashi Koike, Yoshifusa Aizawa

British Journal of Haematology 99 ( 1 ) 222 - 227 1997年

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Blackwell Publishing Ltd

Primary familial and congenital polycythaemia (PFCP) is a rare disease characterized by congenital erythrocytosis inherited in an autosomal dominant fashion. Recently, mutations in the erythropoietin receptor (EpoR) have been identified in PFCP families. We describe a Japanese family with an autosomal dominant inheritance of PFCP. An in vitro colony assay demonstrated hypersensitivity of erythroid progenitors to erythropoietin (Epo) in affected family members. Sequence analysis of RT-PCR products amplified from the C- terminal region of EpoR transcripts in affected family members revealed that they were all heterozygous for C and T bases at position 5986, which suggested a genetic mutation (C to T) on one allele of EpoR. This mutation gave rise to a translation termination codon TAG at amino acid 435. Thus, the resulting EpoR is a truncated protein product lacking all 74 amino acids downstream of the mutation. To date, all genetic mutations affecting a family with PFCP, including this one, have been located in the cytoplasmic negative regulatory region of the EpoR. All mutations gave rise to truncated Epo receptors between Tyrosine 427 and Tyrosine 455. The phosphotyrosines in this region of EpoR have been demonstrated to be binding sites for SHP-1 phosphatase. Therefore PFCP is presumably brought about as a result of genetic mutations which cause the loss of the SFIP-1 binding site in the cytoplasmic region of EpoR.

DOI： 10.1046/j.1365-2141.1997.3583172.x

Profile of cell cycle in hematopoietic malignancy by DNA/RNA quantitation using 7AAD/PY 査読

K. Toba, K. Kishi, T. Koike, E. F. Winton, H. Takahashi, K. Nagai, S. Maruyama, T. Furukawa, S. Hashimoto, M. Masuko, Y. Uesugi, T. Kuroha, N. Tsukada, A. Shibata

Experimental Hematology 24 ( 8 ) 894 - 901 1996年

記述言語：英語掲載種別：研究論文（学術雑誌）

Using 7-amino-actinomycin-D/pyronin Y (7AAD/PY), we analyzed the surface phenotypes and cell cycle of 22 hematopoietic cell lines based on their cellular DNA/RNA content. Populations of G(1a), G(1b), S, and G2M, the DNA index (DI), and the RNA index of S phase (SRI) were calculated by means of DNA/RNA dot plots. Two new parameters were extracted from the cell-cycle profiles: the nucleic acid index of S phase (NI) and the coefficient of variations in the RNA at S phase (SCV). DNA/RNA dot plots of cell lines revealed four characteristic profiles of the cell cycle, defined with the calculated NI and SCV. These were type 0 (small NI, large SCV), type I(small NI, small SCV), type II (large NI, small SCV), and type III (large NI, large SCV). Type 0 included four stem cell lines: one t(1
19) leukemia, two Ph1+ acute lymphocytic leukemia (ALL), and one biphenotypic crisis of chronic granulocytic leukemia (CGL). Type I included five ALL cell lines: three T-ALL and two common B-ALL. Type II contained 10 myeloid cell lines: five AML and five myeloid crisis of CGL. Type III contained three relatively immature lymphoma cell lines: two Burkitt's lymphoma and one follicular center lymphoma. Calculated NI/SCV (%) were as follows: type 0, 2.27 ± 0.19/16.7 ± 3.7
type I, 2.20 ± 0.30/11.1 ± 0.7
type II, 3.64 ± 0.52/11.8 ± 1.0
and type III, 3.60 ± 0.53/17.5 ± 1.9. Cell-cycle analysis of blasts using 7AAD/PY combined with surface phenotyping may yield important information for classifying hematopoietic malignancy within 2 hours of patient admission.