2001 Fiscal Year Annual Research Report

抗P糖蛋白single chain Fv抗体とケモカインの融合蛋白作製と応用研究

Research Project

Project/Area Number	13557053
Research Institution	The University of Tokushima
Principal Investigator	曽根三郎徳島大学, 医学部, 教授 (40145024)
Co-Investigator(Kenkyū-buntansha)	矢野聖二徳島大学, 医学部・附属病院, 講師 (30294672) 西岡安彦徳島大学, 医学部, 講師 (70274199) 谷憲治徳島大学, 医学部, 助教授 (70207166)
Keywords	P糖蛋白 / single chan抗体 / ケモカイン / 薬剤耐性 / 抗体治療 / 免疫療法 / MRK-16 / MCP-1
Research Abstract	がん化学療法に直面する問題点としてがん細胞の薬剤耐性獲得が挙げられる。P糖蛋白質(P-gp)はATPのエネルギーを利用して能動的に抗がん剤の排泄を行うポンプ作用をもち、がん細胞による抗がん剤耐性の獲得に大きく寄与しており、P-gpを標的とした治療が現在試みられている。我々はこれまで、P-gpに対するモノクロナール抗体(MRK-16)が多剤耐性を示す小細胞肺がん細胞の多臓器転移形成に対して治療効果を示すこと、さらに単球の遊走因子であるmonocyte chemoattractant protein-1(MCP-1)遺伝子のがん細胞への導入によりMRK16抗体治療効果が増強することを報告してきた。今回、我々は多剤耐性がん増殖部位へ遊走因子MCP-1を到達させるためのdelivery systemとしてMRK16のsingle chain antibodyの利用に着目した。方法として、MRK-16のH鎖、L鎖それぞれの可変領域をPCR法を用いてクローニングし、それらをリンカーで接続することによりMRK-16 single chain antibody (scFvMRK-16)を作製した。scFvMRK-16のP-gpに対する親和性はモノクローナル抗体であるMRK-16には劣るものの、FACS法を用いてscFvMRK-16がP-gpに結合することを確認した。さらに、mouse MCP-1(JE)遺伝子をクローニングしscFvMRK-16の上流に組み込み、scFvMRK-16とJEのfusion proteinを作製した。今後、このfusion proteinのP-gpへの親和性、JE活性およびマクロファージ遊走能の有無等を検討し、最終的にはin vivoにおいてfusion proteinの多剤耐性がん結節への蓄積性とMRK16抗体との併用による抗腫瘍効果増強作用について検討していく予定である。

Research Products

(21 results)

All Other

All Publications (21 results)

[Publications] Takeuchi, E. et al.: "IL-12-induced production of IL-10 and interferon-gamma by mononuclear cells in lung cancer-associated malignant pleural effusions"Lung Cancer. 35. 171-177 (2002)
[Publications] Shinohara, T. et al.: "NF-kB-dependent expression of metastasis suppressor KAI1/CD82 gene in lung cancer cell lines expressing mutant p53"Cancer Res.. 61. 673-678 (2001)
[Publications] Shinohara, T. et al.: "Transduction of KAI1/CD82 cDNA promotes hematogenous spread of human lung-cancer cells in natural killer cell-depleted SCID mice"Int. J. Cancer. 94. 16-23 (2002)
[Publications] Goto, H. et al.: "Gamma interferon-dependent induction of thymidine phosphorylase/platelet-derived endothelial growth factor through gamma-activated sequence-like element in human macrophages"Cancer Res.. 61. 469-473 (2001)
[Publications] Nishimura, N. et al.: "Enhanced efficiency by centrifugal manupulation of adenovirus-mediated interleukin 12 gene transduction into human monocyte-derived dendritic cells"Hum. Gene Ther.. 12. 333-346 (2001)
[Publications] Nishimura, N. et al.: "Novel centrifugal method for simple and highly efficient adenovirus-mediated green fluorescence protein gene transduction into human monocyte-deried dendritic cells"J. Immunological. Methods.. 253. 113-124 (2001)
[Publications] Takeuchi, E. et al.: "Interleukin (IL-) 15 has less activity than IL-2 to promote type 2 cytokine predominance in tumour-associated mononuclear cells form lung cancer patients"Cytokine. 13. 119-123 (2001)
[Publications] Parajuli, P. et al.: "Humanized anti-ganglioside GM2 antibody is effective to induce antibody-dependent cell-mediated cytotoxicity in mononuclear cells from lung cancer patients"Cancer Lett.. 163. 179-184 (2001)
[Publications] Hanibuchi, M. et al.: "Immunological circumvention of multiple organ metastases of multidrug resistant human small cell lung cancer cells by mouse-human chimeric anti-ganglioside GM2 antibody KM966"Clin. Exp. Metastasis. 18. 353-360 (2001)
[Publications] Cui, P.et al.: "A novel bioactive 31-amino acid endothelin-1 is a potent chemotactic peptide for human neutrophils and monocytes"J. Leukoc. Biol.. 70. 306-312 (2001)
[Publications] Miki, T. et al.: "Bone metastasis model with multi-organ dissemination of human small cell lung cancer (SBC-5) cells in natural killer-cell depleted SCID mice"Oncol. Res.. 12. 209-217 (2001)
[Publications] Tanaka, H. et al.: "CD40 and IFN-gamma dependent T cell activation by human bronchial epithelial cells"J. Med. Invest.. 48. 109-117 (2001)
[Publications] Suzuki, Y. et al.: "Efficient generation of dendritic cells from alveolar and pleural macrophages as well as blood monocytes in patients with lung cancer"Lung Cancer. 34. 195-205 (2001)
[Publications] Ge, N.et al.: "Interferon-gamma activates outwardly rectifying chloride channels in the human bronchial epithelial cell line BEAS-2B"J. Med. Invest.. 48. 97-101 (2001)
[Publications] Sone, S. et al.: "Molecular pathogenesis of lung cancer and its molecular targeted therapy"Internal Med.. 40(2). 167-170 (2001)
[Publications] Huang, L. et al.: "Thrombin promotes fibroblast proliferation during the early stages of experimental radiation pneumonitis"Radiation Res.. 156. 45-52 (2001)
[Publications] Nokihara, H. et al.: "A new quinoline derivative MS-209 reverses multidrug resistance and inhibits multiorgan metastases by P-glycoprotein-expressing human small cell lung cancer cells"Jpn. J. Cancer Res.. 92. 785-792 (2001)
[Publications] Ogushi, F. et al.: "Autoantibodies to IL-1α in sera from rapidly progressive idiopathic pulmonary fibrosis"J. Med. Invest.. 48. 181-189 (2001)
[Publications] Tani, K. et al.: "Protease-induced leukocyte chemotaxis and activation : Roles in host defense and inflammation"J. Med. Invest.. 48. 133-141 (2001)
[Publications] Hase, K. et al.: "Increased CCR4 expression in active systemic lupus erythematosus"J. Leukoc. Biol.. 70. 749-755 (2001)
[Publications] Nishioka, Y. et al.: "Human monocyte-derived and CD83+ blood dendritic cells enhance NK cell-mediated cytotoxicity"Eur. J. Immunol.. 31. 2633-2641 (2001)

2001 Fiscal Year Annual Research Report

抗P糖蛋白single chain Fv抗体とケモカインの融合蛋白作製と応用研究

Principal Investigator

曽根 三郎 徳島大学, 医学部, 教授 (40145024)

Research Products

[Publications] Takeuchi, E. et al.: "IL-12-induced production of IL-10 and interferon-gamma by mononuclear cells in lung cancer-associated malignant pleural effusions"Lung Cancer. 35. 171-177 (2002)

[Publications] Shinohara, T. et al.: "NF-kB-dependent expression of metastasis suppressor KAI1/CD82 gene in lung cancer cell lines expressing mutant p53"Cancer Res.. 61. 673-678 (2001)

[Publications] Shinohara, T. et al.: "Transduction of KAI1/CD82 cDNA promotes hematogenous spread of human lung-cancer cells in natural killer cell-depleted SCID mice"Int. J. Cancer. 94. 16-23 (2002)

[Publications] Goto, H. et al.: "Gamma interferon-dependent induction of thymidine phosphorylase/platelet-derived endothelial growth factor through gamma-activated sequence-like element in human macrophages"Cancer Res.. 61. 469-473 (2001)

[Publications] Nishimura, N. et al.: "Enhanced efficiency by centrifugal manupulation of adenovirus-mediated interleukin 12 gene transduction into human monocyte-derived dendritic cells"Hum. Gene Ther.. 12. 333-346 (2001)

[Publications] Nishimura, N. et al.: "Novel centrifugal method for simple and highly efficient adenovirus-mediated green fluorescence protein gene transduction into human monocyte-deried dendritic cells"J. Immunological. Methods.. 253. 113-124 (2001)

[Publications] Takeuchi, E. et al.: "Interleukin (IL-) 15 has less activity than IL-2 to promote type 2 cytokine predominance in tumour-associated mononuclear cells form lung cancer patients"Cytokine. 13. 119-123 (2001)

[Publications] Parajuli, P. et al.: "Humanized anti-ganglioside GM2 antibody is effective to induce antibody-dependent cell-mediated cytotoxicity in mononuclear cells from lung cancer patients"Cancer Lett.. 163. 179-184 (2001)

[Publications] Hanibuchi, M. et al.: "Immunological circumvention of multiple organ metastases of multidrug resistant human small cell lung cancer cells by mouse-human chimeric anti-ganglioside GM2 antibody KM966"Clin. Exp. Metastasis. 18. 353-360 (2001)

[Publications] Cui, P.et al.: "A novel bioactive 31-amino acid endothelin-1 is a potent chemotactic peptide for human neutrophils and monocytes"J. Leukoc. Biol.. 70. 306-312 (2001)

[Publications] Miki, T. et al.: "Bone metastasis model with multi-organ dissemination of human small cell lung cancer (SBC-5) cells in natural killer-cell depleted SCID mice"Oncol. Res.. 12. 209-217 (2001)

[Publications] Tanaka, H. et al.: "CD40 and IFN-gamma dependent T cell activation by human bronchial epithelial cells"J. Med. Invest.. 48. 109-117 (2001)

[Publications] Suzuki, Y. et al.: "Efficient generation of dendritic cells from alveolar and pleural macrophages as well as blood monocytes in patients with lung cancer"Lung Cancer. 34. 195-205 (2001)

[Publications] Ge, N.et al.: "Interferon-gamma activates outwardly rectifying chloride channels in the human bronchial epithelial cell line BEAS-2B"J. Med. Invest.. 48. 97-101 (2001)

[Publications] Sone, S. et al.: "Molecular pathogenesis of lung cancer and its molecular targeted therapy"Internal Med.. 40(2). 167-170 (2001)

[Publications] Huang, L. et al.: "Thrombin promotes fibroblast proliferation during the early stages of experimental radiation pneumonitis"Radiation Res.. 156. 45-52 (2001)

[Publications] Nokihara, H. et al.: "A new quinoline derivative MS-209 reverses multidrug resistance and inhibits multiorgan metastases by P-glycoprotein-expressing human small cell lung cancer cells"Jpn. J. Cancer Res.. 92. 785-792 (2001)

[Publications] Ogushi, F. et al.: "Autoantibodies to IL-1α in sera from rapidly progressive idiopathic pulmonary fibrosis"J. Med. Invest.. 48. 181-189 (2001)

[Publications] Tani, K. et al.: "Protease-induced leukocyte chemotaxis and activation : Roles in host defense and inflammation"J. Med. Invest.. 48. 133-141 (2001)

[Publications] Hase, K. et al.: "Increased CCR4 expression in active systemic lupus erythematosus"J. Leukoc. Biol.. 70. 749-755 (2001)

[Publications] Nishioka, Y. et al.: "Human monocyte-derived and CD83+ blood dendritic cells enhance NK cell-mediated cytotoxicity"Eur. J. Immunol.. 31. 2633-2641 (2001)

曽根三郎徳島大学, 医学部, 教授 (40145024)