1994 Fiscal Year Annual Research Report

エイズ制圧に向けての基礎研究柱1 HIVの分子遺伝学

Research Project

Project/Area Number	04269104
Research Institution	Kyoto University
Principal Investigator	畑中正一京都大学, ウイルス研究所, 教授 (30142300)
Co-Investigator(Kenkyū-buntansha)	牧正敏名古屋大学, 農学部, 教授 (40183610) 塩田達雄東京大学, 医科学研究所, 助手 (00187329) 志田壽利京都大学, ウイルス研究所, 助教授 (00144395) 足立昭夫京都大学, ウイルス研究所, 助教授 (90127043) 品川日出夫大阪大学, 微生物病研究所, 教授 (40029799)
Keywords	HIV-1 / Gag蛋白 / 感染初期 / EnV / Rev / eIF5A / V3領域 / 細胞宿主域
Research Abstract	大腸菌で産生させ精製したHIV-1のGag先駆体p55は、リコンビナントGag(308-406)-Env(512-611)キメラ蛋白や逆転写酵素と同等の反応性を有する優れた高原であった。これらを用いた高感度診断法を確立した。HIV-2のGag蛋白p26と逆転写酵素の多量生産と精製に成功した(品川)。 HIV-1とHIV-2には合計5種類のアクセサリー遺伝子がある。我々の開発したシステムによる解析から、これらの遺伝子産物が効果をあらわす時期は、Vif,Vpr,Vpr,Nefが感染初期であり、Vpuが感染後期であることがわかった。また、Gag変異体の解析からGagの機能が感染初期と後期の両方に要求されることも判明した(足立)。組換えワクシニアを用いてEnvとCD4により引き起こされる細胞融合を鋭敏に検出する方法を開発した。β-ガラクトシダーゼのαとω相補を利用するものである。この方法によりサイトカラシンDの阻害効果と、融合に関与する細胞成分の存在を示した(志田)。 HIV感染者の体内のHIV-のEnv遺伝子V1領域からV3領域の構造を経時的に解析した。病態の進行した症例では、V3には塩基性荷電が蓄積し、V2には新たな糖鎖付加部位の挿入が認められた。V2の糖鎖付加部位の挿入はV3の塩基性荷電が未だ増加していない症例でも認められた(塩田)。 AIDS遺伝子治療に関する基礎的研究として、HIV-1増殖に必須の調節因子Revタンパク質の機能を強力に抑制するRev変異体を作製し、in vitroにおけるこの変異体の抗ウイルス作用を解析した。Rev変異体遺伝子を恒常的に導入したヒト培養細胞ではHIV-1の増殖ならびに細胞障害性が顕著に抑制された(牧、畑中)。

Research Products
(37 results)

All Other

All Publications (37 results)

[Publications] Kubota, S.: "The origin of human immunodeficiency virus type-1 rev gene:An evolutionary hypothesis" FEBS Lett.338. 118-121 (1994)
[Publications] Ueba, T: "Expression of fibroblast growth factor receptor-1 in human glioma and menigioma tissues" Neurosurgery. 34. 221-226 (1994)
[Publications] El-Farrash, M.A.: "Generation and characterization of a human immunodeficiency virus type 1(HIV-1) mutant resistant to an HIV--1 protease inhibitor" J. Virol.68. 233-239 (1994)
[Publications] Yamada, T.: "The human T-cell leukemia virus type I Tax protein induces apoptosis which is blocked by the Bcl-2 protein" J. Virol.68. 3374-3379 (1994)
[Publications] Yang,H.Q.: "Analysis of calcium-dependent interaction between amino-terminal conserved region of calpastatin functional domain and calmodulin-like domain of μ-calpain large subunit" J. Biol. Chem.269. 18977-18984 (1994)
[Publications] Suzui, H.: "Immunohistochemical reactions for fibroblast growth factor receptor in arteries of patients with moyamoya disease" Neurosurgery. 35. 20-25 (1994)
[Publications] Suzui, H.: "Immunohistochemical study for basic fibroblast growth factor receptor I in pituitary adenomas" Neurosci, Lett.171. 192-196 (1994)
[Publications] Ueba, T.: "Transcriptional regulation of basic fibroblast grwoth factor gene by p53 in human glioblastoma and hepatocellular carcinoma cells" Proc. Natl. Acad. Sci. USA. 91. 9009-9013 (1994)
[Publications] Takano, E.: "Real-time analysis of the calcium-dependent interaction between calmodulin and a synthetic oligopeptide of calcineurin by a surface plasmon resonance biosensor" FEBS Lett.352. 247-250 (1994)
[Publications] Ma, H.: "Amino-terminal conserved region in proteinase inhibitory activity by interacting with calmodulin-like domain of the proteinase" J. Biol. Chem.269. 24430-24436 (1994)
[Publications] Satoh, T.: "Histamine as an activator of cell growth and extracellular matrix reconstruction for human vascular smooth muscle cells" Atherosclerosis. 110. 53-61 (1994)
[Publications] Inoue, H.: "Suppression of the transformed phenotype in hybrids of human T-cell leukemia virus type I tax-transformed rat fibroblasts and normal human fibroblasts" Exp. Cell Res.215. 68-74 (1994)
[Publications] Yamaguchi, R.: "Unphosphorylated and tyrosine phosphorylated forms of a focal adhesion protein,paxillin,are substrates for calpain II in vitro:implications for the possible involvement of calpain II in mitosis-specific degradation of paxillin" FEBS Lett.356. 114-116 (1994)
[Publications] Miyamoto, S.: "Tumor necrosis factor α-induced phosphorylation of IκBα is a signal for its degradation but not dissociation from NF-κB" Proc. Natl. Acad. Sci. USA. 91. 12740-12744 (1994)
[Publications] Iwakura, Y.: "Augmentation of c-fos and c-jun expression in transgenic mice carrying the human T-cell leukemia virus type-I tax gene" Virus Gene. 9. 161-170 (1994)
[Publications] Hirota, K.: "Sensitive enzyme immunoassay (immune complex transfer enzyme immunoassay) for antibodies to reverse transcriptase(RT) of human immunodeficiency virus type 1(HIV-1)using recombinant RT of HIV-1 as antigen" Clin. Chem. Enzym. Comms.6. 175-184 (1994)
[Publications] Hashinaka, K.: "Detection of anti-human immunodeficiency virus type1(HIV-1)immunoglobulin G in urine by an ultrasensitive enzyme immunoassay (immune complex transfer enzyme immunoassay) with recombinant reverse transcriptase as an antigen" J. Clin. Microbiol.32. 819-822 (1994)
[Publications] Hashida, S.: "Detection of antibody IgG to HIV-1 in urine by ultrasensitive enzyme immunoassay (immune complex tranfer eyzyme immunoassay)using recombinant p24as antigne for diagnosis of HIV-1 infection" J. Clin. Lab. Anal.8. 86-95 (1994)
[Publications] Hashida, S.: "Diagnosis of HIV-1 infection by detection of antibody IgG to HIV-1 in urine with ultrasensitive enzyme immunoassay(immune complex transfer enzyme immunoassay)using recombinant proteins as antigens" J. Clin. Lab. Anal.8. 237-246 (1994)
[Publications] Hashinaka, K.: "Conjugation of recombinant reverse transcriptase of HIV-1 to β-D-galactosidase from Escherichia coli for ultrasensitive enzyme immunoassay(immune complex transfer enzyme immunoassay of anti-HIV-1 IgG" J. Immunol. Meth.172. 179-187 (1994)
[Publications] Nakagawa, Y.: "Inhibition of murine AIDS(MAIDS)development by the transplantation of bone marrow cells carrying the Fv-4 resistant gene to MAIDS virus-infected mice" J. Virol.68. 1438-1441 (1994)
[Publications] Kawamura, M.: "Growth ability of human immunodeficiency virus type 1 auxiliary gene mutants in primary blood macrophage cultures" J. Gen. Virol.75. 2427-2431 (1994)
[Publications] Igarashi, T.: "Persistent infection with SIVmac chimeric virus having tat,rev vpu,env,and nef of HIV-1 in macaque Pmonkeys" AIDS Res. Human Retroviruses. 10. 1021-1029 (1994)
[Publications] Kawamura, M.: "Human immunodeficiency virus Vpx is required for the early phase of replication in peripheral blood mononuclear cells" Microbiol. Immunol.38. 871-878 (1994)
[Publications] Katahira, J.: "Effect of translation initiation factor elF-5A on HTLV-1 Rex and HIV Rev functions that had been inhibited trans-dominantly a Rex mutant deficient in RNA binding" J. Virol.(in press).
[Publications] Adachi, A..: "Methods for HIV-SIV gene analysis" Methods Mol. Genet.(in press).
[Publications] Kannagi, M.: "Cytotoxic T cell response and expression of the target antigen in HTLV-1 infection" Leukemia. 8. 54-59 (1994)
[Publications] Jin, N-Y.: "Nigh level of expression by vaccinia virus vectors based on combination effect of promoters of A-type inclusion body,tandemly repeated mutant 7.5-KDa protein and hemagglutinin genes" Arch. Virol.138. 315-330 (1994)
[Publications] Tanaka, Y.: "Induction of antibody responses that neutralize human T-cell leukemia virus type I infection in vitro and in vivo by peptide immunization" J. Virol.68. 6323-6331 (1994)
[Publications] Katahira, J.: "A cellular protein which associates with HTLV-1 Rex protein in the presence of the target mRNA" Oncogene. 9. 3535-3544 (1994)
[Publications] Furukawa, K.: "Clonal expansion of CD8+ cytotoxic T lymphocytes against human T cell lymphotropic virus type I (HTLV-1)genome products in HTLV-1 assoiciated myelopathy/tropical spastic paraparesis patients" J. Clin. Invest.94. 1830-1839 (1994)
[Publications] Ohnishi, Y.: "A furin-defective cell line is able to process correctly the gp160 of human immunodeficiency virus type 1" J. Virol.68. 4075-4079 (1994)
[Publications] Oka, S.: "Genetic analysis of HTV-1 during rapid progression to AIDS in an apparently healthy man" AIDS Res. Human Retroviruses. 10. 271-277 (1994)
[Publications] Shinoda, T.: "A naturally occurring single basic amino acid substitution in the V3 region of the human immunodeficiency virus type 1 Env protein alters the cellular host range and antigenic structure of the virus" J. Virol.68. 7689-769619GA35:Furuta, R.A. (1994)
[Publications] "Use of a human immunodeficiency virus type 1 Rev mutant without nucleolar dysfunction as a candidate for potential AIDS therapy" J. Virol.69. 1591-1599 (1995)
[Publications] 畑中正一: "ATLとAIDS" 南江堂, 150 (1994)
[Publications] Gordon, S.: "The Legacy of Cell Fusion" Oxford Science Publications, 295 (1994)

1994 Fiscal Year Annual Research Report

エイズ制圧に向けての基礎研究 柱1 HIVの分子遺伝学

Principal Investigator

畑中 正一 京都大学, ウイルス研究所, 教授 (30142300)

Research Products

[Publications] Kubota, S.: "The origin of human immunodeficiency virus type-1 rev gene:An evolutionary hypothesis" FEBS Lett.338. 118-121 (1994)

[Publications] Ueba, T: "Expression of fibroblast growth factor receptor-1 in human glioma and menigioma tissues" Neurosurgery. 34. 221-226 (1994)

[Publications] El-Farrash, M.A.: "Generation and characterization of a human immunodeficiency virus type 1(HIV-1) mutant resistant to an HIV--1 protease inhibitor" J. Virol.68. 233-239 (1994)

[Publications] Yamada, T.: "The human T-cell leukemia virus type I Tax protein induces apoptosis which is blocked by the Bcl-2 protein" J. Virol.68. 3374-3379 (1994)

[Publications] Yang,H.Q.: "Analysis of calcium-dependent interaction between amino-terminal conserved region of calpastatin functional domain and calmodulin-like domain of μ-calpain large subunit" J. Biol. Chem.269. 18977-18984 (1994)

[Publications] Suzui, H.: "Immunohistochemical reactions for fibroblast growth factor receptor in arteries of patients with moyamoya disease" Neurosurgery. 35. 20-25 (1994)

[Publications] Suzui, H.: "Immunohistochemical study for basic fibroblast growth factor receptor I in pituitary adenomas" Neurosci, Lett.171. 192-196 (1994)

[Publications] Ueba, T.: "Transcriptional regulation of basic fibroblast grwoth factor gene by p53 in human glioblastoma and hepatocellular carcinoma cells" Proc. Natl. Acad. Sci. USA. 91. 9009-9013 (1994)

[Publications] Takano, E.: "Real-time analysis of the calcium-dependent interaction between calmodulin and a synthetic oligopeptide of calcineurin by a surface plasmon resonance biosensor" FEBS Lett.352. 247-250 (1994)

[Publications] Ma, H.: "Amino-terminal conserved region in proteinase inhibitory activity by interacting with calmodulin-like domain of the proteinase" J. Biol. Chem.269. 24430-24436 (1994)

[Publications] Satoh, T.: "Histamine as an activator of cell growth and extracellular matrix reconstruction for human vascular smooth muscle cells" Atherosclerosis. 110. 53-61 (1994)

[Publications] Inoue, H.: "Suppression of the transformed phenotype in hybrids of human T-cell leukemia virus type I tax-transformed rat fibroblasts and normal human fibroblasts" Exp. Cell Res.215. 68-74 (1994)

[Publications] Yamaguchi, R.: "Unphosphorylated and tyrosine phosphorylated forms of a focal adhesion protein,paxillin,are substrates for calpain II in vitro:implications for the possible involvement of calpain II in mitosis-specific degradation of paxillin" FEBS Lett.356. 114-116 (1994)

[Publications] Miyamoto, S.: "Tumor necrosis factor α-induced phosphorylation of IκBα is a signal for its degradation but not dissociation from NF-κB" Proc. Natl. Acad. Sci. USA. 91. 12740-12744 (1994)

[Publications] Iwakura, Y.: "Augmentation of c-fos and c-jun expression in transgenic mice carrying the human T-cell leukemia virus type-I tax gene" Virus Gene. 9. 161-170 (1994)

[Publications] Hirota, K.: "Sensitive enzyme immunoassay (immune complex transfer enzyme immunoassay) for antibodies to reverse transcriptase(RT) of human immunodeficiency virus type 1(HIV-1)using recombinant RT of HIV-1 as antigen" Clin. Chem. Enzym. Comms.6. 175-184 (1994)

[Publications] Hashinaka, K.: "Detection of anti-human immunodeficiency virus type1(HIV-1)immunoglobulin G in urine by an ultrasensitive enzyme immunoassay (immune complex transfer enzyme immunoassay) with recombinant reverse transcriptase as an antigen" J. Clin. Microbiol.32. 819-822 (1994)

[Publications] Hashida, S.: "Detection of antibody IgG to HIV-1 in urine by ultrasensitive enzyme immunoassay (immune complex tranfer eyzyme immunoassay)using recombinant p24as antigne for diagnosis of HIV-1 infection" J. Clin. Lab. Anal.8. 86-95 (1994)

[Publications] Hashida, S.: "Diagnosis of HIV-1 infection by detection of antibody IgG to HIV-1 in urine with ultrasensitive enzyme immunoassay(immune complex transfer enzyme immunoassay)using recombinant proteins as antigens" J. Clin. Lab. Anal.8. 237-246 (1994)

[Publications] Hashinaka, K.: "Conjugation of recombinant reverse transcriptase of HIV-1 to β-D-galactosidase from Escherichia coli for ultrasensitive enzyme immunoassay(immune complex transfer enzyme immunoassay of anti-HIV-1 IgG" J. Immunol. Meth.172. 179-187 (1994)

[Publications] Nakagawa, Y.: "Inhibition of murine AIDS(MAIDS)development by the transplantation of bone marrow cells carrying the Fv-4 resistant gene to MAIDS virus-infected mice" J. Virol.68. 1438-1441 (1994)

[Publications] Kawamura, M.: "Growth ability of human immunodeficiency virus type 1 auxiliary gene mutants in primary blood macrophage cultures" J. Gen. Virol.75. 2427-2431 (1994)

[Publications] Igarashi, T.: "Persistent infection with SIVmac chimeric virus having tat,rev vpu,env,and nef of HIV-1 in macaque Pmonkeys" AIDS Res. Human Retroviruses. 10. 1021-1029 (1994)

[Publications] Kawamura, M.: "Human immunodeficiency virus Vpx is required for the early phase of replication in peripheral blood mononuclear cells" Microbiol. Immunol.38. 871-878 (1994)

[Publications] Katahira, J.: "Effect of translation initiation factor elF-5A on HTLV-1 Rex and HIV Rev functions that had been inhibited trans-dominantly a Rex mutant deficient in RNA binding" J. Virol.(in press).

[Publications] Adachi, A..: "Methods for HIV-SIV gene analysis" Methods Mol. Genet.(in press).

[Publications] Kannagi, M.: "Cytotoxic T cell response and expression of the target antigen in HTLV-1 infection" Leukemia. 8. 54-59 (1994)

[Publications] Jin, N-Y.: "Nigh level of expression by vaccinia virus vectors based on combination effect of promoters of A-type inclusion body,tandemly repeated mutant 7.5-KDa protein and hemagglutinin genes" Arch. Virol.138. 315-330 (1994)

[Publications] Tanaka, Y.: "Induction of antibody responses that neutralize human T-cell leukemia virus type I infection in vitro and in vivo by peptide immunization" J. Virol.68. 6323-6331 (1994)

[Publications] Katahira, J.: "A cellular protein which associates with HTLV-1 Rex protein in the presence of the target mRNA" Oncogene. 9. 3535-3544 (1994)

[Publications] Furukawa, K.: "Clonal expansion of CD8+ cytotoxic T lymphocytes against human T cell lymphotropic virus type I (HTLV-1)genome products in HTLV-1 assoiciated myelopathy/tropical spastic paraparesis patients" J. Clin. Invest.94. 1830-1839 (1994)

[Publications] Ohnishi, Y.: "A furin-defective cell line is able to process correctly the gp160 of human immunodeficiency virus type 1" J. Virol.68. 4075-4079 (1994)

[Publications] Oka, S.: "Genetic analysis of HTV-1 during rapid progression to AIDS in an apparently healthy man" AIDS Res. Human Retroviruses. 10. 271-277 (1994)

[Publications] Shinoda, T.: "A naturally occurring single basic amino acid substitution in the V3 region of the human immunodeficiency virus type 1 Env protein alters the cellular host range and antigenic structure of the virus" J. Virol.68. 7689-769619GA35:Furuta, R.A. (1994)

[Publications] "Use of a human immunodeficiency virus type 1 Rev mutant without nucleolar dysfunction as a candidate for potential AIDS therapy" J. Virol.69. 1591-1599 (1995)

[Publications] 畑中 正一: "ATLとAIDS" 南江堂, 150 (1994)

[Publications] Gordon, S.: "The Legacy of Cell Fusion" Oxford Science Publications, 295 (1994)

エイズ制圧に向けての基礎研究柱1 HIVの分子遺伝学

畑中正一京都大学, ウイルス研究所, 教授 (30142300)

[Publications] 畑中正一: "ATLとAIDS" 南江堂, 150 (1994)