1997 Fiscal Year Annual Research Report

マウスモノクローナル抗体及びそのヒト型化抗体を用いた新しい遺伝子導入法の開発

Research Project

Project/Area Number	08457363
Research Institution	Osaka University
Principal Investigator	清水惠司大阪大学, 医学部, 助教授 (50162699)
Co-Investigator(Kenkyū-buntansha)	池中一裕岡崎国立共同研究機構, 生理学研究所, 教授 (00144527) 宮尾泰慶大阪大学, 医学部・付属病院, 医員
Keywords	Monoclonal antibody / Humanized antibody / Single-chained antibody / Gene therapy / Gene transfer / HVJ-liposome / Immunoliposome / Internalization
Research Abstract	我々が作製したONS-M21マウスモノクローナル抗体(Br J Cancer 68:831-837,1993)が認識するグリオーマ関連抗原はintegrin α3であることが同定された(Br J Cancer,in revised)。現在、この抗原がグリオーマ細胞上に発現するメカニズムについて研究中である。また、この抗体のヒト型化や(Mol Immunol 32:407-416,1995)、さらには可変領域のみからなる単鎖抗体も開発できている(Jpn J Cancer Res,in submitted)。^<125>I標識ONS-M21抗体あるいはそのヒト型化抗体をヌードラットの尾静脈より投与すると、24時間後には、各臓器や血中より脳内移植腫瘍への集積が高くなることが実証された。さらに、ヒト型化抗体を用いたオートラジオグラフでは、MRIで診断できなかった実験脳腫瘍も描出可能であった(Eur J Nucl Med,in submitted)。また、このONS-M21抗体、そのヒト型化や単鎖抗体が細胞膜上の腫瘍関連抗原と結合すると、細胞内によく取り込まれることから(Cancer Lett,in press)、殺細胞遺伝子導入や、免疫複合体を用いたバイオターゲッティング法への応用が期待されている。しかし、この抗体を用いた殺細胞遺伝子導入は、導入を試みた分子量が大き過ぎるので実現不可能であり、遺伝子分割導入を現在試みている。一方、我々は、Polyoma ori発現遺伝子をパッケージング細胞に導入することで、高力価な脳特異的レトロウイルスベクターを開発した(Hum Gene Ther 9:161-172,1998)。そして、このマウスMBPプロモータで制御されたamphotropicレトロウイルスベクターによって、ヒトグリオーマ細胞が強く抑制されることも証明した(Jpn J Cancer Res 88:678-686,1997)。さらに、レトロウイルスレセプターを保持しないグリオーマに対しては、このヒト型化抗体とHVJリポソームの組み合わせによるグリオーマ特異的イムノリポソーム法の応用を計画している(Gene Ther 4:768-772,1997)。

Research Products
(18 results)

All Other

All Publications (18 results)

[Publications] Moriuchi S: "An Immunohistochemical Analysis of Medulloblastoma and PNET with Emphasis on N-myc Protein Expression." Anticancer Res. 16. 2687-2692 (1996)
[Publications] Moriuchi S: "Decreased N-myc expression in human medulloblastoma cell lines during differentiation." Anticancer Res. 17. 301-306 (1997)
[Publications] Tamura M: "Targeted killing of migrating glioma cells by injection of HTK-modified glioma cells." Hum Gene Ther. 8. 381-391 (1997)
[Publications] Miyao Y: "Usefulness of a mouse myelin basic protein promoter for gene therapy of malignant glioma : myelin basic protein promoter is strongly active in human malignant glioma cells." Jpn J Cancer Res. 88. 678-686 (1997)
[Publications] Mabuchi E: "Gene delivery by HVJ-liposome in the experimental gene therapy of murine glioma." Gene Ther. 4. 768-772 (1997)
[Publications] Miyao Y: "FT-175,a Synthetic Inhibitor of the Complement Pathway,Protects against the Inactivation of Infectious Retrovirus by Human Serum." Hum Gene Ther. 8. 1575-1583 (1997)
[Publications] Kishima H: "Possible common origin of α-fetoprotein-and human chorionic gonadotropin-secreting cells in intracranial germ cell tumor." J Neurosurg. 88. 576-580 (1998)
[Publications] Park KC: "Interferon yield MHC antigen expression of human medulloblastoma cells and its suppression during dibutyryl cyclic AMP-induced differentiation :" Cell Mol Neurobiol. (in press). (1998)
[Publications] Yamada M: "In vitro study on intrathecal use of 5-fluoro-2'-deoxyuridine (FdUrd) for meningeal dissemination of malignant brain tumors." J Neuro-oncol. (in press). (1998)
[Publications] Park KC: "Regulation of interferon responses in medulloblastoma cells by interferon regulatory factor -1 and -2." Br J Cancer. (in press). (1998)
[Publications] Shimizu K: "Internalization with high targeting potential of mouse monoclonal antibody ONS-M21 recognizing human malignant glioma antigen." Cancer lett. (in press). (1998)
[Publications] Kishima H: "Systemic interleukin-12 displays antitumor activity in the mouse central nervous system." Br J Cancer. (in press). (1998)
[Publications] Yamaguchi Y: "Myelin proteolipid protein (PLP),but not DM-20,is an inositol hexakisphosphate binding protein." J Biol Chem. 271. 27838-27846 (1996)
[Publications] Ikeda H: "Retroviral introduction of the p16 gene into murine cell lines to elicit marked antiproliferative effects." Jpn.J Cancer Res. 88. 712-717 (1997)
[Publications] Baba H: "GFAP gene expression during development of astrocyte." Dev Neurosci. 19. 49-57 (1997)
[Publications] Morita N: "Astrocytic lineage analysis by detection of GFAP promoter activity in vitro." Dev Neurosci. 19. 210-218 (1997)
[Publications] Yoshimatsu T: "Improvement of retroviral packaging cell lines by introducing the polymavirus early region." Hum Gene Ther. 9. 161-172 (1998)
[Publications] Miyao Y: "Brain Tumor Research and Therapy" Nagai M (ed),Springer-Verlag Tokyo,(Japan), 456 (1996)

1997 Fiscal Year Annual Research Report

マウスモノクローナル抗体及びそのヒト型化抗体を用いた新しい遺伝子導入法の開発

Principal Investigator

清水 惠司 大阪大学, 医学部, 助教授 (50162699)

Research Products

[Publications] Moriuchi S: "An Immunohistochemical Analysis of Medulloblastoma and PNET with Emphasis on N-myc Protein Expression." Anticancer Res. 16. 2687-2692 (1996)

[Publications] Moriuchi S: "Decreased N-myc expression in human medulloblastoma cell lines during differentiation." Anticancer Res. 17. 301-306 (1997)

[Publications] Tamura M: "Targeted killing of migrating glioma cells by injection of HTK-modified glioma cells." Hum Gene Ther. 8. 381-391 (1997)

[Publications] Miyao Y: "Usefulness of a mouse myelin basic protein promoter for gene therapy of malignant glioma : myelin basic protein promoter is strongly active in human malignant glioma cells." Jpn J Cancer Res. 88. 678-686 (1997)

[Publications] Mabuchi E: "Gene delivery by HVJ-liposome in the experimental gene therapy of murine glioma." Gene Ther. 4. 768-772 (1997)

[Publications] Miyao Y: "FT-175,a Synthetic Inhibitor of the Complement Pathway,Protects against the Inactivation of Infectious Retrovirus by Human Serum." Hum Gene Ther. 8. 1575-1583 (1997)

[Publications] Kishima H: "Possible common origin of α-fetoprotein-and human chorionic gonadotropin-secreting cells in intracranial germ cell tumor." J Neurosurg. 88. 576-580 (1998)

[Publications] Park KC: "Interferon yield MHC antigen expression of human medulloblastoma cells and its suppression during dibutyryl cyclic AMP-induced differentiation :" Cell Mol Neurobiol. (in press). (1998)

[Publications] Yamada M: "In vitro study on intrathecal use of 5-fluoro-2'-deoxyuridine (FdUrd) for meningeal dissemination of malignant brain tumors." J Neuro-oncol. (in press). (1998)

[Publications] Park KC: "Regulation of interferon responses in medulloblastoma cells by interferon regulatory factor -1 and -2." Br J Cancer. (in press). (1998)

[Publications] Shimizu K: "Internalization with high targeting potential of mouse monoclonal antibody ONS-M21 recognizing human malignant glioma antigen." Cancer lett. (in press). (1998)

[Publications] Kishima H: "Systemic interleukin-12 displays antitumor activity in the mouse central nervous system." Br J Cancer. (in press). (1998)

[Publications] Yamaguchi Y: "Myelin proteolipid protein (PLP),but not DM-20,is an inositol hexakisphosphate binding protein." J Biol Chem. 271. 27838-27846 (1996)

[Publications] Ikeda H: "Retroviral introduction of the p16 gene into murine cell lines to elicit marked antiproliferative effects." Jpn.J Cancer Res. 88. 712-717 (1997)

[Publications] Baba H: "GFAP gene expression during development of astrocyte." Dev Neurosci. 19. 49-57 (1997)

[Publications] Morita N: "Astrocytic lineage analysis by detection of GFAP promoter activity in vitro." Dev Neurosci. 19. 210-218 (1997)

[Publications] Yoshimatsu T: "Improvement of retroviral packaging cell lines by introducing the polymavirus early region." Hum Gene Ther. 9. 161-172 (1998)

[Publications] Miyao Y: "Brain Tumor Research and Therapy" Nagai M (ed),Springer-Verlag Tokyo,(Japan), 456 (1996)

清水惠司大阪大学, 医学部, 助教授 (50162699)