1992 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	Osaka University
Principal Investigator	中田篤男大阪大学, 微生物病研究所, 教授 (80029769)
Co-Investigator(Kenkyū-buntansha)	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)
Keywords	HIV-1の分子遺伝学 / Gagタンパク質と抗体検出 / Vifとウイルス粒子の成熟促進 / Vpuとウイルス粒子の放出促進 / ワクシニア・ベクターとenvの発現 / Envタンパク質の抗原性 / RNA-PCR法によるV3領域の増幅 / V3領域の変異と宿主域の拡大
Research Abstract	本研究班は、HIVを分子遺伝学的に解析して、その増殖機構を明らかにし、抗HIV療法の開発を目標としている。本年度の研究実積は以下の通りである。中田は、HIVの各遺伝子を大腸菌で発現させて遺伝子産物(タンパク質)を精製し、研究用、診断用、さらにはワクチン開はを目指しているが、gag領域をプロセスされない形で発現させてp55(未プロセス産物)を精製し、そのプロセス産物であるp17,p24およびp15の4種類のタンパク質と感染者および非感染者の血清との免疫学的反応を調ベ、p55が感染者血清と特異的に最も強い反応を示すことを明らかにした。足立は、HIV-1の一段増殖系を作成してHIVの非必須遺伝子の機能を調べた。vifは感染初期のウイルス粒子の成熟を促進し、vpuは感染後期に細胞からのウイルス粒子の放出を促進することを証明した。また、両遺伝子およびvpr,vpxの作用が細胞の種類に依存して特異的に発現することも明らかにした。志田は、プロモーターを改良したワクシニア・ベクターを作成し、cat遺伝子とHIVのenv領域を組み込んで発現を調べた。各タンパク質の細胞当たりの生産量が、バキュロ・ウイルスをベクターとしたときと同レベルに上昇することを確認した。また、このenv組換え体ウイルスの感染初期に発現したEnvタンパク質が高い免疫原性を示すことも明らかにした。塩田は、HIV感染者の血清中のenvのV3領域をRNA-PCR法で増幅して、V3領域を交換した組換え体ウイルスを作成して解析した。感染初期と後期の血清中に存在したV3をもつ組換え体ウイルスの細胞宿主域を比較し、同一感染者の病状の進行に伴ったウイルスの細胞宿主域の拡大とV3領域に生じた変異との相関関係を示した。この変異の誘発機構を明らかにしようとしている。

Research Products

(20 results)

All Other

All Publications (20 results)

[Publications] Saitoh,A.,Tanaka,N.,Nakata,A.,Ikuta,K.and Shinagawa,H.: "A Unique Monoclonal Antibody That Recognizes Mature o17 of HIV-1 but Not Its Precursor" Microbiol.Immunol.36. 105-111 (1992)
[Publications] Tanaka,N.,Saitoh,A.,Nakata,A.and Shinagawa H.: "A Sinple Method for Overproduction and Purification of p24 Gag Protein of Human Immunodeficiency Virus Type 1" Microbiol.Immunol.36. 823-831 (1992)
[Publications] Sakai,H.,Sakuragi,J.,Sakuragi,S.,Shibata,R.and Adachi,A.: "Functional analysis of biologically distinct genetic variants of simian immunodeficiency virus isolated from a mandrill." Virology. 189. 161-166 (1992)
[Publications] Sakuragi,J.,Sakai,H.,Sakurage,S.,Shibata,R.,Wain-Hobson,S.,Hayami,M., and Adachi,A.: "Functional classification of simian immunodeficiency virus isolated from a chimpanzee by transactivators." Virology. 189. 354-358 (1992)
[Publications] Sakai,H.,Sakuragi,J.,Sakuragi,S.,Shivata,R.,Hayami,M.,Ishimoto,A.and Adachi,A.: "Genetic characterization of simian immunodeficiency virus esolated from an African mandrill." Arch.Virol.125. 1-14 (1992)
[Publications] Tomonaga,K.,Norimine,J.,Shin,Y.S.,Fukasawa,M.,Miyazawa,T.,Adachi,A.,Toyosaki,T.,Kawaguchi Y.,Kai,C.a: "Identification of afeline immunodeficiency virus gene which is essential for cell-free virus infectivity." J.Virol.66. 6181-6185 (1992)
[Publications] Sakuragi S,Shibata,R.,Mukai,R.,Komatsu,T.,Fukasawa,M.,Sakai,H.,Sakuragi,J.,Kawamura,M., ibuki,K.,Hayami,M.and Adachi.A.: "Infection of macaque monkeys with a chimeric human and simian immunodeficiency virus." J.Gen.Virol.73. 2983-2987 (1992)
[Publications] Sakai,H.,Sakuragi,S.,Sakuragi,J.,Kawamura,M.,Shibata,R.and Adachi,A.: "Sequences responsible for efficient replication of simian immunodeficiency virus SIVmnd in cells of the monocyte/macrophage Iineage." J.Gen.Virol.73. 2989-2993 (1992)
[Publications] Sakai,H.,Kawamura,M.,Sakuragi,J.,SAkuragi,S.,Shibata,R.,Ishimoto.A.,Ono,N.,Ueda,S. and Adachi,A.: "Integration is essential for efficient gene expression of human immunodeficiency virus type 1." J.Virol.67. (1993)
[Publications] Sakai,H.,SHibata,R.,Sakuragi,J.,Sakuragi,S.,Kawamura,M.and Adachi,A.: "Cell-dependent requirement of human immunodeficiency virus type 1 vif protein for maturation of virus particles." J.Virol.67. (1993)
[Publications] Sakai,H.,Sakuragi,J.,Sakuragi,S.,Kawamura,M.and Adachi A.: "Compatibility of tat and rev transactivators in the primate lentiviruses." Arch.Virol.(1993)
[Publications] Adachi,A.,Sakuragi,S.,Kawamura,M.and Sakai,H.: "Genetic analysis of primate immunodeficiency viruses." Methods in Molecular Genetics. (1993)
[Publications] Miyazawa,T.,Kohmoto,M.,Kawaguchi,Y.,Tomonaga,K.,Touosaki,T.,Ikuta,K.,Adachi,A.and Mikami,T.: "The AP-1 binding site in the feline immunodeficiency virus long terrninal repeat is not requered for virus replication in feline T lymphocytes." J.Gen.Virol.(1993)
[Publications] Tanaka,T.,Masuda,M.,Yoshida,A.,Shida,H.,Nyunoya,H.,Shimotohno,K.and Tozawa,H.: "Antigenic structure of the transactivator protein encoded by human T-cell leukemia virus type-1(HTLV-1),as defined by a panel of monoclonal antibodies." AIDS Res.Human Retroviruses. 8. 227-235 (1992)
[Publications] Taguchi,F.,Ikeda,T.and Shida,H.: "Molecular cloning and expression of a spike protein of neurovirulent munne corona virus JHMV variant c1-2." J.Gen.Virol.73. 1065-1072 (1992)
[Publications] Kannagi,M.,Shida,H.,Igarashi,H.,Maruyama,I.,Hattori,T.,Osame,M.,Inoko,H.and Harada,S.: "Target epitope in the tax protein of human T cell leukemia virus type I recognized by class I major histocompatibility complex-restricted cytotoxic T cell." J.Virol.66. 2928-2933 (1992)
[Publications] Iwakura,Y.,Shioda,T.,Tosu,M.,Yoshida,E.,Havashi,M.,Nagata,T.,and Shibuta,H.: "The induction of cataracts by HIV-1 in transgenic mice." AIDS. 6. 1069-1075 (1992)
[Publications] Shioda,T.,Levy,J.A.,and Cheng-Mayer,C.: "Small amino acid changes in the V3 hyper-variable region of gp120 can affect T-cell line and macrophage tropisms of HIV-1." Proc.Natl.Acad.Sci.USA. 89. 9434-9438 (1992)
[Publications] Hayashi,T.,Shioda,T.,Iwakura,Y.,and Shibuta,H.: "RNA packaging signal of human immunodeficiency virus type 1." Virology. 188. 590-599 (1992)
[Publications] 志田壽利: "蛋白質核酸酵素" 共立出版, 6 (1992)

1992 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

中田 篤男 大阪大学, 微生物病研究所, 教授 (80029769)

Research Products

[Publications] Saitoh,A.,Tanaka,N.,Nakata,A.,Ikuta,K.and Shinagawa,H.: "A Unique Monoclonal Antibody That Recognizes Mature o17 of HIV-1 but Not Its Precursor" Microbiol.Immunol.36. 105-111 (1992)

[Publications] Tanaka,N.,Saitoh,A.,Nakata,A.and Shinagawa H.: "A Sinple Method for Overproduction and Purification of p24 Gag Protein of Human Immunodeficiency Virus Type 1" Microbiol.Immunol.36. 823-831 (1992)

[Publications] Sakai,H.,Sakuragi,J.,Sakuragi,S.,Shibata,R.and Adachi,A.: "Functional analysis of biologically distinct genetic variants of simian immunodeficiency virus isolated from a mandrill." Virology. 189. 161-166 (1992)

[Publications] Sakuragi,J.,Sakai,H.,Sakurage,S.,Shibata,R.,Wain-Hobson,S.,Hayami,M., and Adachi,A.: "Functional classification of simian immunodeficiency virus isolated from a chimpanzee by transactivators." Virology. 189. 354-358 (1992)

[Publications] Sakai,H.,Sakuragi,J.,Sakuragi,S.,Shivata,R.,Hayami,M.,Ishimoto,A.and Adachi,A.: "Genetic characterization of simian immunodeficiency virus esolated from an African mandrill." Arch.Virol.125. 1-14 (1992)

[Publications] Tomonaga,K.,Norimine,J.,Shin,Y.S.,Fukasawa,M.,Miyazawa,T.,Adachi,A.,Toyosaki,T.,Kawaguchi Y.,Kai,C.a: "Identification of afeline immunodeficiency virus gene which is essential for cell-free virus infectivity." J.Virol.66. 6181-6185 (1992)

[Publications] Sakuragi S,Shibata,R.,Mukai,R.,Komatsu,T.,Fukasawa,M.,Sakai,H.,Sakuragi,J.,Kawamura,M., ibuki,K.,Hayami,M.and Adachi.A.: "Infection of macaque monkeys with a chimeric human and simian immunodeficiency virus." J.Gen.Virol.73. 2983-2987 (1992)

[Publications] Sakai,H.,Sakuragi,S.,Sakuragi,J.,Kawamura,M.,Shibata,R.and Adachi,A.: "Sequences responsible for efficient replication of simian immunodeficiency virus SIVmnd in cells of the monocyte/macrophage Iineage." J.Gen.Virol.73. 2989-2993 (1992)

[Publications] Sakai,H.,Kawamura,M.,Sakuragi,J.,SAkuragi,S.,Shibata,R.,Ishimoto.A.,Ono,N.,Ueda,S. and Adachi,A.: "Integration is essential for efficient gene expression of human immunodeficiency virus type 1." J.Virol.67. (1993)

[Publications] Sakai,H.,SHibata,R.,Sakuragi,J.,Sakuragi,S.,Kawamura,M.and Adachi,A.: "Cell-dependent requirement of human immunodeficiency virus type 1 vif protein for maturation of virus particles." J.Virol.67. (1993)

[Publications] Sakai,H.,Sakuragi,J.,Sakuragi,S.,Kawamura,M.and Adachi A.: "Compatibility of tat and rev transactivators in the primate lentiviruses." Arch.Virol.(1993)

[Publications] Adachi,A.,Sakuragi,S.,Kawamura,M.and Sakai,H.: "Genetic analysis of primate immunodeficiency viruses." Methods in Molecular Genetics. (1993)

[Publications] Miyazawa,T.,Kohmoto,M.,Kawaguchi,Y.,Tomonaga,K.,Touosaki,T.,Ikuta,K.,Adachi,A.and Mikami,T.: "The AP-1 binding site in the feline immunodeficiency virus long terrninal repeat is not requered for virus replication in feline T lymphocytes." J.Gen.Virol.(1993)

[Publications] Tanaka,T.,Masuda,M.,Yoshida,A.,Shida,H.,Nyunoya,H.,Shimotohno,K.and Tozawa,H.: "Antigenic structure of the transactivator protein encoded by human T-cell leukemia virus type-1(HTLV-1),as defined by a panel of monoclonal antibodies." AIDS Res.Human Retroviruses. 8. 227-235 (1992)

[Publications] Taguchi,F.,Ikeda,T.and Shida,H.: "Molecular cloning and expression of a spike protein of neurovirulent munne corona virus JHMV variant c1-2." J.Gen.Virol.73. 1065-1072 (1992)

[Publications] Kannagi,M.,Shida,H.,Igarashi,H.,Maruyama,I.,Hattori,T.,Osame,M.,Inoko,H.and Harada,S.: "Target epitope in the tax protein of human T cell leukemia virus type I recognized by class I major histocompatibility complex-restricted cytotoxic T cell." J.Virol.66. 2928-2933 (1992)

[Publications] Iwakura,Y.,Shioda,T.,Tosu,M.,Yoshida,E.,Havashi,M.,Nagata,T.,and Shibuta,H.: "The induction of cataracts by HIV-1 in transgenic mice." AIDS. 6. 1069-1075 (1992)

[Publications] Shioda,T.,Levy,J.A.,and Cheng-Mayer,C.: "Small amino acid changes in the V3 hyper-variable region of gp120 can affect T-cell line and macrophage tropisms of HIV-1." Proc.Natl.Acad.Sci.USA. 89. 9434-9438 (1992)

[Publications] Hayashi,T.,Shioda,T.,Iwakura,Y.,and Shibuta,H.: "RNA packaging signal of human immunodeficiency virus type 1." Virology. 188. 590-599 (1992)

[Publications] 志田 壽利: "蛋白質核酸酵素" 共立出版, 6 (1992)

エイズ制圧に向けての基礎研究柱1.HIVの分子遺伝学

中田篤男大阪大学, 微生物病研究所, 教授 (80029769)

[Publications] 志田壽利: "蛋白質核酸酵素" 共立出版, 6 (1992)