2001 Fiscal Year Annual Research Report

HIVライフサイクルに必須な細胞因子の同定および新しい抗HIV薬の標的探索

Research Project

Project/Area Number	13226027
Research Category	Grant-in-Aid for Scientific Research on Priority Areas (C)
Research Institution	Tokyo Medical and Dental University
Principal Investigator	山本直樹東京医科歯科大学, 大学院・医歯学総合研究科, 教授 (00094053)
Co-Investigator(Kenkyū-buntansha)	山岡昇司東京医科歯科大学, 大学院・医歯学総合研究科, 助教授 (90263160)
Keywords	AIDS / HIV-1 / 増殖 / 細胞因子 / レセプター / コレセプター / CXCR-4
Research Abstract	従来の抗HIVの治療標的であるウイルス側酵素と異なり、ウイルスの細胞内での複製及びその病原性(細胞致死効果)に必要な細胞側因子を同定することで、新しい治療標的の探索をめざす。さらに動物感染モデルの作製を試みる。もう一方のアプローチとして、すでに同定されているユニークな細胞側因子であるHIVのコレセプターを標的とし、しかも経口投与可能な低分子アンタゴニストを開発する。 1)HIV侵入後に生存可能な変異細胞の樹立の試みを行った。ウイルスの細胞侵入を100%保証し感染効率をモニターするために、HIV遺伝子のenv領域に欠損を持ち、nef部分にEGFP遺伝子を組み込んで、VSV-Gでpseudotypeとしたウイルスを293T細胞で作製した。このウイルスをヒトT細胞株に高M.O.I.で感染させると感染細胞が数日間で完全に死滅することがわかったので、あらかじめHIV-1高感受性ヒトT細胞株、MT-4細胞、にフレームシフト変異誘起剤ICR-191で複数回処理した細胞プールにウイルスを作用させたのち、生存細胞を増殖させる試みを行った。これにより、大多数の細胞が死滅する中、少数の生き残りを観察することが出来、ここまでは予定通りであった。この後、ウイルスへの抵抗性を確認するためとウイルスの侵入もれによる細胞生存を除外する目的で、このウイルス感染をさらに数回行ってなお生存する細胞を樹立する試みた。今のところ理由は不明であるが非常に高いM.O.I.でも感染を免れる細胞が存在することが問題となっている。そこで現在は細胞数の快復を待たずに頻回のウイルスchallengeを行い、変異細胞のclonesを分離する試みを行っているところである。本法では研究の特色・独創性の項でも述べたように、HIV-1耐性clonesの分離はそのまま研究全体の成功をもかなりの部分で約束することになるので、現在その分離に全力を傾注しているところである。 2)KRH-1636をリード化合物とした多数の誘導体の化学合成を行い、試験管内の抗HIV-1アッセイを実施。その詳細なメカニズムの解析を行った。 CXCR4アンタゴニストとして、すでに見出しているKRH-1636をリード化合物としてその周辺物質の誘導体の化学合成を行い、将来の実用化を想定した薬物の絞り込みをおこなった。

Research Products

(18 results)

All Other

All Publications (18 results)

[Publications] R.Tanaka: "Unique monoclonal antibody recognizing the third extracellular loop of CXCR4 induces lymphocyte agglutination and enhances human immunodeficiency virus type 1-mediated syncytium formation and productive infection"J Virol.. 75. 11534-11543 (2001)
[Publications] H.Tamamura: "Development of specific CXCR4 inhibitors possessing high selectivity indexes as well as complete stability in serum based on an anti-HIV peptide T140. Bioorg"Bioorg Med Chem Lett.. 11. 1897-1902 (2001)
[Publications] H.Tamamura: "Synthesis and evaluation of bifunctional anti-HIV agents based on specific CXCR4 antagonists-AZT conjugation"Bioorg Med Chem Lett.. 9. 2179-2187 (2001)
[Publications] H.Tamamura: "Conformational study of a highly specific CXCR4 inhibitor, T140, disclosing the close proximity of its intrinsic pharmacophores associated with strong anti-HIV activity"Bioorg Med Chem Lett.. 11. 359-362 (2001)
[Publications] M.Takahoko: "Infectious DNA clone of HIV type 1 A/G recombinant (CRF02_AG) replicable in peripheral blood mononuclear cells"AIDS Res Hum Retroviruses. 17. 1083-1087 (2001)
[Publications] Y.Takahashi: "OX40 Stimulation by gp34/OX40 Ligand Enhances Productive Human Immunodeficiency Virus Type 1 Infection"J Virol.. 75. 6748-6757 (2001)
[Publications] K.Takahashi: "Two basic regions of NCp7 are sufficient for conformational conversion of HIV-1 dimerization initiation site from kissing-loop dimer to extended-duplex dimer"J Biol Chem.. 276. 31274-31278 (2001)
[Publications] N.Mori: "Induction of monocyte chemoattractant protein 1 by Helicobacter pylori involves NF-kappaB"Infect Immun.. 69. 1280-1286 (2001)
[Publications] N.Mori: "Human T-cell leukemia virus type I oncoprotein Tax represses Smad-dependent transforming growth factor beta signaling through interaction with CREB-binding protein/p300"Blood. 97. 2137-2144 (2001)
[Publications] Y.Miura: "Critical contribution of tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) to apoptosis of human CD4+ T cells in HIV-1-infected hu-PBL-NOD-SCID mice"J Exp Med.. 193. 651-660 (2001)
[Publications] S.Kusagawa: "Isolation and characterization of a full-length molecular DNA clone of Ghanaian HIV type 1 intersubtype A/G recombinant CRF02 AG, which is replication competent in a restricted host range"AIDS Res. Hum Retroviruses. 17. 649-655 (2001)
[Publications] S.A.Jones: "The soluble interleukin 6 receptor : mechanisms of production and implications in disease"Faseb J.. 15. 43-58 (2001)
[Publications] K.Iwai: "Human T-cell leukemia virus type 1 tax protein activates transcription through AP-1 site by inducing DNA binding activity in T cells"Virology. 279. 38-46 (2001)
[Publications] K.Ishikawa: "Genetic analysis of HIV type 2 from Ghana and Guinea-Bissau, West Africa"AIDS Res Hum Retroviruses. 17. 1661-1663 (2001)
[Publications] S.M.Hurst: "II-6 and its soluble receptor orchestrate a temporal switch in the pattern of leukocyte recruitment seen during acute inflammation"Immunity. 14. 705-714 (2001)
[Publications] R.Handema: "Emergence of New HIV-1 Subtypes Other Than Subtype C among Antenatal Women in Lusaka, Zambia"AIDS Res Hum Retroviruses. 17. 759-763 (2001)
[Publications] E.Baba: "Functional CD4 T cells after intercellular molecular transfer of OX40 ligand"J Immunol. 167. 875-883 (2001)
[Publications] N.Mori: "Human T-cell leukemia virus type I Tax transactivates the matrix metalloproteinase-9 gene : potential role in mediating adult T-cell leukemia invasiveness"Virology. 99. 1341-1349 (2002)

2001 Fiscal Year Annual Research Report

HIVライフサイクルに必須な細胞因子の同定および新しい抗HIV薬の標的探索

Principal Investigator

山本 直樹 東京医科歯科大学, 大学院・医歯学総合研究科, 教授 (00094053)

Research Products

[Publications] R.Tanaka: "Unique monoclonal antibody recognizing the third extracellular loop of CXCR4 induces lymphocyte agglutination and enhances human immunodeficiency virus type 1-mediated syncytium formation and productive infection"J Virol.. 75. 11534-11543 (2001)

[Publications] H.Tamamura: "Development of specific CXCR4 inhibitors possessing high selectivity indexes as well as complete stability in serum based on an anti-HIV peptide T140. Bioorg"Bioorg Med Chem Lett.. 11. 1897-1902 (2001)

[Publications] H.Tamamura: "Synthesis and evaluation of bifunctional anti-HIV agents based on specific CXCR4 antagonists-AZT conjugation"Bioorg Med Chem Lett.. 9. 2179-2187 (2001)

[Publications] H.Tamamura: "Conformational study of a highly specific CXCR4 inhibitor, T140, disclosing the close proximity of its intrinsic pharmacophores associated with strong anti-HIV activity"Bioorg Med Chem Lett.. 11. 359-362 (2001)

[Publications] M.Takahoko: "Infectious DNA clone of HIV type 1 A/G recombinant (CRF02_AG) replicable in peripheral blood mononuclear cells"AIDS Res Hum Retroviruses. 17. 1083-1087 (2001)

[Publications] Y.Takahashi: "OX40 Stimulation by gp34/OX40 Ligand Enhances Productive Human Immunodeficiency Virus Type 1 Infection"J Virol.. 75. 6748-6757 (2001)

[Publications] K.Takahashi: "Two basic regions of NCp7 are sufficient for conformational conversion of HIV-1 dimerization initiation site from kissing-loop dimer to extended-duplex dimer"J Biol Chem.. 276. 31274-31278 (2001)

[Publications] N.Mori: "Induction of monocyte chemoattractant protein 1 by Helicobacter pylori involves NF-kappaB"Infect Immun.. 69. 1280-1286 (2001)

[Publications] N.Mori: "Human T-cell leukemia virus type I oncoprotein Tax represses Smad-dependent transforming growth factor beta signaling through interaction with CREB-binding protein/p300"Blood. 97. 2137-2144 (2001)

[Publications] Y.Miura: "Critical contribution of tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) to apoptosis of human CD4+ T cells in HIV-1-infected hu-PBL-NOD-SCID mice"J Exp Med.. 193. 651-660 (2001)

[Publications] S.Kusagawa: "Isolation and characterization of a full-length molecular DNA clone of Ghanaian HIV type 1 intersubtype A/G recombinant CRF02 AG, which is replication competent in a restricted host range"AIDS Res. Hum Retroviruses. 17. 649-655 (2001)

[Publications] S.A.Jones: "The soluble interleukin 6 receptor : mechanisms of production and implications in disease"Faseb J.. 15. 43-58 (2001)

[Publications] K.Iwai: "Human T-cell leukemia virus type 1 tax protein activates transcription through AP-1 site by inducing DNA binding activity in T cells"Virology. 279. 38-46 (2001)

[Publications] K.Ishikawa: "Genetic analysis of HIV type 2 from Ghana and Guinea-Bissau, West Africa"AIDS Res Hum Retroviruses. 17. 1661-1663 (2001)

[Publications] S.M.Hurst: "II-6 and its soluble receptor orchestrate a temporal switch in the pattern of leukocyte recruitment seen during acute inflammation"Immunity. 14. 705-714 (2001)

[Publications] R.Handema: "Emergence of New HIV-1 Subtypes Other Than Subtype C among Antenatal Women in Lusaka, Zambia"AIDS Res Hum Retroviruses. 17. 759-763 (2001)

[Publications] E.Baba: "Functional CD4 T cells after intercellular molecular transfer of OX40 ligand"J Immunol. 167. 875-883 (2001)

[Publications] N.Mori: "Human T-cell leukemia virus type I Tax transactivates the matrix metalloproteinase-9 gene : potential role in mediating adult T-cell leukemia invasiveness"Virology. 99. 1341-1349 (2002)

山本直樹東京医科歯科大学, 大学院・医歯学総合研究科, 教授 (00094053)