FUNCTIONAL AND DNA ALALYSIS OF THE NOVEL PROTEIN (R21 PROTEIN) INVOLVED IN MEMBRANE FUSION
FUNCTIONAL AND DNA ALALYSIS OF THE NOVEL PROTEIN (R21 PROTEIN) INVOLVED IN MEMBRANE FUSION
批准号:
10670416
负责人:
NAKAMURA Minoru
金额:
$2.3万
依托单位:
依托单位国家:
日本
项目类别:
Grant-in-Aid for Scientific Research (C)
财政年份:
1998
资助国家:
日本
项目状态:
已结题
起止时间:
1998 至 1999
中文摘要
By immunizing rats with cocultured HTLV-I-positive ILT8M2 and HTLV-I-negative MOLT-4cells,we isolated one monoclonal antibody(MAb),designated as mAb R21,which enhances the syncytium formation induced by coculturing ILT8M2 cells with MOLT-4cells.Since the enhancing activity by mAb R21 of syncytium formation was observed only in the presence of a factor contained in fetal calf serum(FCS)which seems to bind to mAb R21,we purified this serum factor from FCS using a mAb R21-coupled Sepharose 4B column.The partial amino acid sequence of the purified protein indicates that R21 protein is a novel bovine serum protein which has approximately 90%amino acid homology with bovine platelet factor4,a member of CXC chemokine family.Next,to study the mechanism of viral entry and cell fusion mediated by HTLV-I env.Glycoproteins,and to identify the HTLV-I cellular receptor(S),we constructed a cell fusion-dependent reporter gene activation assay and a vesicular stomatitis virus(VSV)pse…More udotype assay.The cell fusion-dependent reporter gene activation assay,in which T7RNA polymerase in donor cells co-expressing env glycoproteins activates a luciferase gene in recipient cells upon cell fusion,revealed that a broad range of cells are susceptible to HTLV-I env-mediated cell fusion。This indicated that cell-to-cell transmission of HTLV-I can occur in a wider range of cells than previously reported.The VSV pseudotype assay,which consists of recombinant VSV containing the green fluorescent protein gene instead of the receptor-binding G protein gene(VSV G I i D1*i D 1)and complemented with HTLV-I env glycoproteins(VSV G I i D 1*I e D 1-Env),revealed that VSV G I i D 1*E D 1-Env infects efficiently only HepG 2 and 293T cells,indicating that cell-free G I D 1*E D 1-Env infects efficiently only HepG 2 and 293T cells,The characterization of the HTLV-I receptor(S)by various chemical modifications of HepG2 and 293T cells indicated that some glycosaminoglycans and phospholipids but not proteins on the cell surface may play a major role in HTLV-I env-mediated viral entry。The expression cloning of HTLV-I receptor is now underway using cDNA library from HepG2 cells.Less:Less
英文摘要
By immunizing rats with cocultured HTLV-I-positive ILT8M2 and HTLV-I-negative MOLT-4 cells, we isolated one monoclonal antibody (mAb), designated as mAb R21, which enhances the syncytium formation induced by coculturing ILT8M2 cells with MOLT-4 cells. Since the enhancing activity by mAb R21 of syncytium formation was observed only in the presence of a factor contained in fetal calf serum(FCS) which seems to bind to mAb R21, we purified this serum factor from FCS using a mAb R21- coupled Sepharose 4B column. The partial amino acid sequence of the purified protein indicates that R21 protein is a novel bovine serum protein which has approximately 90% amino acid homology with bovine platelet factor 4, a member of CXC chemokine family.Next, to study the mechanism of viral entry and cell fusion mediated by HTLV-I env. glycoproteins, and to identify the HTLV-I cellular receptor(s), we constructed a cell fusion-dependent reporter gene activation assay and a vesicular stomatitis virus (VSV) pse … More udotype assay. The cell fusion-dependent reporter gene activation assay, in which T7 RNA polymerase in donor cells co-expressing env glycoproteins activates a luciferase gene in recipient cells upon cell fusion, revealed that a broad range of cells are susceptible to HTLV-I env-mediated cell fusion. This indicated that cell-to-cell transmission of HTLV-I can occur in a wider range of cells than previously reported. The VSV pseudotype assay, which consists of recombinant VSV containing the green fluorescent protein gene instead of the receptor-binding G protein gene(VSV△GィイD1*ィエD1) and complemented with HTLV-I env glycoproteins (VSV△GィイD1*ィエD1-Env), revealed that VSV△GィイD1*ィエD1-Env infects efficiently only HepG2 and 293T cells, indicating that cell-free HTLV-I transmission is less efficient than cell-to-cell transmission. The characterization of the HTLV-I receptor(s) by various chemical modifications of HepG2 and 293T cells indicated that some glycosaminoglycans and phospholipids but not proteins on the cell surface may play a major role in HTLV-I env-mediated viral entry. The expression cloning of HTLV-I receptor is now underway using cDNA library from HepG2 cells. Less
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Shinji Shimoda,Judy V.D.Water,Aftab Ansari,Minoru Nakamura et al: "Identification and precursorfrequency analysis of a common T cell epitope motif in mitochondrial autoantigens in primary biliary cirrhosis." J.Clin.Invest.102. 1831-1840 (1998)
Shinji Shimoda、Judy V.D.Water、Aftab Ansari、Minoru Nakamura 等人:“原发性胆汁性肝硬化线粒体自身抗原中常见 T 细胞表位基序的鉴定和前体频率分析。”
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通讯作者:
Ishibashi, H., Shimoda, S., Shigematsu, H., Ichiki, Y., Nakamura, M., Hayashida, K., and Gershwin, E.M.: "Immune pathophysiology of primary biliary cirrhosis. Progress in Hepatology - Liver and Immunology"Volume5. edited by M. Yamanaka, G. Toda, and T. Ta
Ishibashi, H.、Shimoda, S.、Shigematsu, H.、Ichiki, Y.、Nakamura, M.、Hayashida, K. 和 Gershwin, E.M.:“原发性胆汁性肝硬化的免疫病理生理学。肝病学进展 - 肝脏和免疫学
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B.Kitae,K.Usuku,Y.Yamamoto,S.Yashiki,M.Nakamura,et al: "Human CD4^+ T lymphocytes recognize a highly conserved epitope of human T lymphotropic virus type I(HTLD-1)env gp21 restricted by HLADRB^*0101." Clin.Exp.Immunol.111. 278-285 (1998)
B.Kitae、K.Usuku、Y.Yamamoto、S.Yashiki、M.Nakamura 等人:“人类 CD4^ T 淋巴细胞识别人类 T 淋巴细胞病毒 I 型 (HTLD-1)env gp21 的高度保守表位,受
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K. Okuma, M. Nakamura, S. Nakano, and Y. Niho.: "Identification of a novel bovine serum protein which is involved in human T-cellleukemiavirus type I(HTLV-I)-induced syncytium formation."Arch. Virol.. 144. 1-18 (1999)
K. Okuma、M. Nakamura、S. Nakano 和 Y. Niho.:“鉴定一种新型牛血清蛋白,该蛋白参与人 T 细胞白血病病毒 I 型 (HTLV-I) 诱导的合胞体形成。”Arch。
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Shinji Shimoda, Minoru Nakamura, Hirohisa Shigematsu, Hironori Tanimoto, Toshihumi Gushima, and Hiromi Ishibashi.: "Mimicrypeptides of human PDC-E2 163-176 peptide the immunodominantT cell epitope of primary biliary cirrhosis."Hepatology. (in press). (200
Shinji Shimoda、Minoru Nakamura、Hirohisa Shigematsu、Hironori Tanimoto、Toshihumi Gushima 和 Hiromi Ishibashi。:“人 PDC-E2 163-176 肽的模拟肽,原发性胆汁性肝硬化的免疫显性 T 细胞表位。”肝病学。
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共 9 条
Genome-wide associationstudy to detect disease-associated genes in Japanese patients with primary biliary cirrhosis
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批准号:23591006
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项目类别:Grant-in-Aid for Scientific Research (C)
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资助金额:$3.41万
-
财政年份:2011
-
负责人:NAKAMURA Minoru
-
依托单位:
New clinical classification and the criteria for predicting long-term outcome in primary biliary cirrhosis
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批准号:20590800
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项目类别:Grant-in-Aid for Scientific Research (C)
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资助金额:$3.16万
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财政年份:2008
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负责人:NAKAMURA Minoru
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依托单位:
Identification of new molecular targets and the study for the mechanism of bile ducts-and hepatocyte-destruction in primary biliary cirrhosis
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批准号:17590696
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项目类别:Grant-in-Aid for Scientific Research (C)
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资助金额:$2.24万
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财政年份:2005
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负责人:NAKAMURA Minoru
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依托单位:
Construction of antibody-library to identify etiology-associated-antigen
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批准号:15591075
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项目类别:Grant-in-Aid for Scientific Research (C)
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资助金额:$2.24万
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财政年份:2003
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负责人:NAKAMURA Minoru
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依托单位:
Identification and characterization of T cell epitope in primary biliary cirrhosis
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批准号:11694287
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项目类别:Grant-in-Aid for Scientific Research (B)
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资助金额:$5.12万
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财政年份:1999
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负责人:NAKAMURA Minoru
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依托单位:
Identification of HTLV-1 neutralizing epitopes and the development of an HTLV-1 peptide based vaccine
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批准号:04670391
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项目类别:Grant-in-Aid for General Scientific Research (C)
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资助金额:$1.34万
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财政年份:1992
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负责人:NAKAMURA Minoru
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依托单位:
海外基金