Generation and application of fully human antibodies to a new tumor-associated antigen, Ep-CAM

• Project/Area Number: 18590544
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Laboratory medicine
• Research Institution: Fukuoka University
• Principal Investigator: KUROKI Masahide Fukuoka University, Faculyt of Medicine, Professor (40122692)
• Co-Investigator(Kenkyū-buntansha): KINUGASA Tetsushi Fukuoka University, Faculty of Medicine, Senior Assistant Professor (90279266) ; SHIBAGUCHI Hirotomo Fukuoka University, Faculty of Medicine, Senior Assistant Professor (60295061)
• Project Period (FY): 2006 – 2007
• Project Status: Completed (Fiscal Year 2007)
• Budget Amount: ¥3,980,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥480,000); Fiscal Year 2007: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000); Fiscal Year 2006: ¥1,900,000 (Direct Cost: ¥1,900,000)
• Keywords: cancer / immunology / biotechnology / monoclonal antibody / Ep-CAM / ADCC / CDC

Research Abstract

There are numerous tumor-associated antigens currently used for the diagnosis and treatment of cancer. However, no tumor-associated antigen can always be an exact and complete target molecule, because almost all of them are not absolutely tumor-specific. In the present study, we prepared human mAbs against a pan-carcinoma antigen, Ep-CAM, using a genetically engineered mouse (KM mouse^<TM>) that contains the human immunoglobulin genes. Spleen cells from KM mice immunized with recombinant Ep-CAM were fused with the P3-U1 mouse myeloma cells. Of 44 anti-Ep-CAM clones analyzed, two were of IgG4 and the others of IgM clones. The two IgG4 clones were suggested to recognize the same antigenic determinant or two closely located determinants. The two IgG4 and one of the IgM clones tested revealed antibody-dependent cell-mediated cytotoxicity and complement-dependent cytotoxicity, respectively, against Ep-CAM-expressing cells in vitro. It is also known that IgG4 hardly binds to the FcγIII receptor, which involves in ADCC by NK cells. In the presence of the IgG4 clone M13-13, however, a slight increase was observed in the cytotoxicity of IL-2-activated PBMC against tumor cells. IgG4 antibodies are able to bind to the FcγI receptor on monocytes and thereby exert ADCC. The ADCC activity observed with M13-13 might be due to activated monocytes. Therefore, we engineered our IgM clone M11-29 to convert it to the IgG1 type using its V-region genes so that it can exhibit both ADCC and CDC to Ep-CAM-expressing tumor cells. The V-region genes obtained were also useful to prepare single chain Fv antibodies to retarget cylotoxic T cells or a retrovector to tumor cells. Taken together, these data suggest that these fully human mAbs produced against Ep-CAM and their V-region genes, which are applicable for preparation of engineered antibody fragments, may be useful for antibody-based therapy of cancer.

Research Products

• [Journal Article] Correlation between lymph node metastasis and expression of VEGF-C,VEGF-D and VEGFR-3 in T1 lung adenocarcinoma (2007)
  • Author(s): Maekawa, S., Iwasaki, A., Enatsu, S., Kawakami, T., Kuroki, Mo., et. al.
  • Journal Title: Anticancer Res. 27 Pages: 3755-3741
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] Tumor targeting carboxylesterase fused with anti CEA scFv improve the anticancer effect with a less toxic dose of irinotecan (2007)
  • Author(s): Uchino, J., Takayama, K., Harada, A., Sone, T., et. al.
  • Journal Title: Cancer Gene Ther. 15 Pages: 94-100
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] Sonodynamic therapy of cancer using a novel porphyrin derivative,DCPH-P-Na(I),which is devoid of photosensitivity (2007)
  • Author(s): Hachimine, K., Shibaguchi, H., Kuroki, Mo., Yamada, H., et. al.
  • Journal Title: Cancer Sci. 98 Pages: 916-920
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] Correlation between lymph node metastasis and expression of TEGF-C, VEGF-D and VEGFR-3 in T1 lung adenocarcinoma (2007)
  • Author(s): Maekawa, S., Iwasaki, A., Enatsu, S., Kawakami, T., Kuroki, Mo., et. al.
  • Journal Title: Anticancer Res. 27 Pages: 3755-3741
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Tumor targeting carboxylesterase fused with anti CEA scFv improve the anticancer effect with a less toxic dose of irinotecan (2007)
  • Author(s): Uchino, J., Takayama, K., Harada, A., Sone, T., Harada, T., et. al.
  • Journal Title: Cancer Gene Ther 15 Pages: 94-100
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Sonodynamic therapy of cancer using a novel porphyrin derivative, DCPH-P-Na (I), which is devoid of photosensitivity (2007)
  • Author(s): Hachimine, K., Shibaguchi, H., Kuroki, Mo., Yamada, H., et. al.
  • Journal Title: Cancer Sci. 98 Pages: 916-920
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Correlation between lymph node metastasis and expression of VEGF-C,VEGF-D and VEGFR-3 in T1 lung adenocarcinoma (2007)
  • Author(s): Maekawa, S., Iwasaki, A., Enatsu, S., Kawakami, T., Kuroki, Mo, et. al.
  • Journal Title: Anticancer Res 27 Pages: 3755-3741
  • Peer Reviewed
• [Journal Article] Tumor targeting carboxylesterase fused withanti CEA scFv improve the anticancer effect with a less toxic dose of irinotecan (2007)
  • Author(s): Uchino, J., Takayama, K., Harada, A., Sone, T., Harada, T., et. al.
  • Journal Title: Cancer Gene Ther 15 Pages: 94-100
  • Peer Reviewed
• [Journal Article] Sonodynamic therapy of cancer using a novel porphyrin derivative,DCPH-PNa(I),which is devoid of photosensitivity (2007)
  • Author(s): Hachimine, K., Shibaguchi, H., Kuroki, Mo., Yamada, H., et. al.
  • Journal Title: Cancer Sci 98 Pages: 916-920
  • Peer Reviewed
• [Journal Article] Prognostic value of carcinoma-associated antigen MK-1 in urinary bladdercarcinoma (2006)
  • Author(s): EL-Meghawry EL-Kenawy, A., EL-Kott, A.F., et. al.
  • Journal Title: Int.J.Bio.Markers 21 Pages: 170-174
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] MK-1 expression in carcinoma of the ampulla of Vater as a predictor of improved prognosis after surgical resection (2006)
  • Author(s): Hamada, Y., Nakayama, Y., Mizoguchi, M., et. al.
  • Journal Title: Cancer Lett. 243 Pages: 211-216
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] CEA-targeted selective gene therapy for gastric cancer through FZ33 fiber modified adenovirus vector (2006)
  • Author(s): Tanaka, T., Huang, J., Hirai, S., Kuroki, Mo., et. al.
  • Journal Title: Clin.Cancer Res. 12 Pages: 3803-3813
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] Possible applications of antibodies or their genes in cancer therapy (2006)
  • Author(s): Kuroki, M., Huang, J., Shibaguchi, H., Zhao, J., et. al.
  • Journal Title: Anticancer Res. 26 Pages: 4019-4026
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] Antitumor activity of chimeric immunoreceptor gene-modified Tc1 and Th1 cells against autologous CEA-expressing colon cancer cells (2006)
  • Author(s): Sasaki, T., Ikeda, H., Sato, M., Ohkuri, T., Abe, H., et. al.
  • Journal Title: Cancer Sci. 97 Pages: 920-927
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] Prognostic value of carcinoma-associated antigen MK-1 in urinary bladder carcinoma (2006)
  • Author(s): EL-Meghawry EL-Kenawy, A., EL-Kott, A. F., et. al.
  • Journal Title: Int. J. Bio. Markers 21 Pages: 170-174
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] CEA-targeted selective gene therapy for gastric cancer through FZ33 fiber modified adenovirus vector (2006)
  • Author(s): Tanaka, T., Huang, J., Hirai, S., Kuroki, Mo., et. al.
  • Journal Title: Clin. Cancer Res. 12 Pages: 3803-3813
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Antitumor activity of chimeric immunoreceptor gene-modified Tc1 and Th1 cells against autologous CEA expressing colon cancer cells (2006)
  • Author(s): Sasaki, T., Ikeda, H., Sato, M., Ohkuri, T., Abe, H., et. al.
  • Journal Title: Cancer Sci. 97 Pages: 920-927
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Prognostic value of carcinoma-associated antigen MK-1 in urinary bladder carcinoma (2006)
  • Author(s): EL-Meghawry, EL-Kenawy, A., EL-Kott, A.F.et al.
  • Journal Title: Int J Biol.Markers 21・3 Pages: 171-171
• [Journal Article] Possible applications of antibodies or their genes in cancer (2006)
  • Author(s): Kuroki, M, Huang, J, Shibaguchi, H, Zhao, J.et al.
  • Journal Title: Anticancer Res Cancer Res. 26・6 Pages: 4019-4026
• [Journal Article] CEA-targeted slective gene therapy for gastric cancer through FZ33 fiber modified adenovirus vector (2006)
  • Author(s): Tanakja, T, Huang, J, Hirai, S, kuroki, Mo.et al.
  • Journal Title: Clin.Cancer Res. 12・12 Pages: 3803-3813
• [Journal Article] Antitumor activity of chimeric immunoreceptor gene-modified Tel and Thl cells against autologous CEA-expressing colon cancer cells (2006)
  • Author(s): Sasaki, T, Ikeda, H, Sato, M, Ohkuri, T, Abe, H.et al.
  • Journal Title: Cancer Sci. 97・9 Pages: 920-927
• [Journal Article] IgG isotype conversion of a novel human anti-carcinoembonic antigen antibody to increase its biological activity (2006)
  • Author(s): Huang, J, Shibaguchi, H, Zhao, J, Luo, N.et al.
  • Journal Title: Anticancer Res. 26・2 Pages: 1057-1064
• [Journal Article] MK-1 expression in carcinoma of the ampulla of Vater as a predictor of improved prognosis after surgical resection (2006)
  • Author(s): Hamada, Y, Nakayama, Y, Mizoguchi, M, Ikeda, S.et al.
  • Journal Title: Cancer Lett. 243・2 Pages: 211-216
• [Presentation] HB-EGF or amphiregulin is a promising target for human cancer therapy (2007)
  • Author(s): 四元 房典, 八木 裕史, 園田 顕三, 辻岡 筧, 蜂須 賀徹, ほか
  • Organizer: 第66回日本癌学会学術総会
  • Place of Presentation: 横浜
  • Year and Date: 2007-10-05
  • Description: 「研究成果報告書概要(和文)」より
• [Presentation] Selective up-regulation of claudin-1 and claudin-2 in colorectal cancer (2007)
  • Author(s): 衣笠 哲史, 黒木 求, 芝口 浩智, 田中 俊裕, ほか
  • Organizer: 第66回日本癌学会学術総会
  • Place of Presentation: 横浜
  • Year and Date: 2007-10-05
  • Description: 「研究成果報告書概要(和文)」より
• [Presentation] HB-EGF or amphiregulin is a promising target for human cancer therapy nation and endocytosis (2007)
  • Author(s): Yotsumoto, F., Yagi, H., Sonoda, K., Tsujioka, H., Hachisuga, T., et. al.
  • Organizer: 66th Ann. Meet. of Jpn Cancer Assoc.
  • Place of Presentation: Yokohama
  • Year and Date: 2007-10-05
  • Description: 「研究成果報告書概要(欧文)」より
• [Presentation] Selective up-regulation of claudin-1 and claudin-2 in colorectal cancer (2007)
  • Author(s): Kinugasa, T., Kuroki, M., Shibaguchi, H., Tanaka, T., et. al.
  • Organizer: 66th Ann. Meet. of Jpn Cancer Assoc.
  • Place of Presentation: Yokohama
  • Year and Date: 2007-10-05
  • Description: 「研究成果報告書概要(欧文)」より
• [Presentation] Amphiregulin enchances drug-resistance for pancreatic and colon cancers (2007)
  • Author(s): 深見 達弥, 八木 裕史, 四元 房典, 沖 英次, 黒木 政秀, ほか
  • Organizer: 第66回日本癌学会学術総会
  • Place of Presentation: 横浜
  • Year and Date: 2007-10-04
  • Description: 「研究成果報告書概要(和文)」より
• [Presentation] Evaluation of Adv-FZ33 mediated gene transduction among anti-CEA Abs with different subclasses of mouse and human IgGs (2007)
  • Author(s): 田中 俊裕, 趙 君, 黒木 求, 衣笠 哲史, 芝口 浩智, 加藤 和則, ほか
  • Organizer: 第66回日本癌学会学術総会
  • Place of Presentation: 横浜
  • Year and Date: 2007-10-04
  • Description: 「研究成果報告書概要(和文)」より
• [Presentation] Amphiregulin enhances drug-resistance for pancreatic and colon cancers. (2007)
  • Author(s): Fukami, T., Yagi, H., Yotsumoto, F., Oki, E., Kuroki, M., et. al.
  • Organizer: 66th Ann. Meet. of Jpn Cancer Assoc.
  • Place of Presentation: Yokohama
  • Year and Date: 2007-10-04
  • Description: 「研究成果報告書概要(欧文)」より
• [Presentation] Evaluation of Adv-FZ33 mediated gene transduction among anti-CEA Abs with different subclasses of mouse and human IgGs (2007)
  • Author(s): Tanaka, T., Zhao, J., Kuroki, M., Kinugasa, T., et. al.
  • Organizer: 66th Ann. Meet. of Jpn Cancer Assoc.
  • Place of Presentation: Yokohama
  • Year and Date: 2007-10-04
  • Description: 「研究成果報告書概要(欧文)」より
• [Presentation] Expression patterns of tight junction associated proteins in colon cancer (2006)
  • Author(s): 衣笠 哲史, 黒木 求, 芝口 浩智, 山田 博美, ほか
  • Organizer: 第65回日本癌学会学術総会
  • Place of Presentation: 横浜
  • Year and Date: 2006-09-30
  • Description: 「研究成果報告書概要(和文)」より
• [Presentation] Pleural lavage cytology before and after lung resection in non-small cell Lung cancer patients (2006)
  • Author(s): 江夏 総太郎, 吉田 純司, 横瀬 智之, 西村 光世, 西脇 裕, ほか
  • Organizer: 第65回日本癌学会学術総会
  • Place of Presentation: 横浜
  • Year and Date: 2006-09-30
  • Description: 「研究成果報告書概要(和文)」より
• [Presentation] Correlation between lymph node metastasis and expression of VEGF-C,-D And VEGFR-3 in primary lung adenocarcinomas (2006)
  • Author(s): 前川 信一, 岩崎 昭憲, 江夏 総太郎, 衣笠 哲史, 黒木 求, ほか
  • Organizer: 第65回日本癌学会学術総会
  • Place of Presentation: 横浜
  • Year and Date: 2006-09-30
  • Description: 「研究成果報告書概要(和文)」より
• [Presentation] Expression patterns of tight junction associated proteins in colon cancer (2006)
  • Author(s): Kinugasa, T., Kuroki, M., Shibaguchi, H., et. al.
  • Organizer: 65th Ann. Meet. of Jpn Cancer Assoc.
  • Place of Presentation: Yokohama
  • Year and Date: 2006-09-30
  • Description: 「研究成果報告書概要(欧文)」より
• [Presentation] Pleural lavage cytology before and after lung resection in non-small cell Lung cancer patients (2006)
  • Author(s): Enatsu, S., Yoshida, J., Yokose, T., Nishimura, M., Nishiwaki, Y., et. al.
  • Organizer: 65th Ann. Meet. of Jpn Cancer Assoc.
  • Place of Presentation: Yokohama
  • Year and Date: 2006-09-30
  • Description: 「研究成果報告書概要(欧文)」より
• [Presentation] Correlation between lymph node metastasis and expression of VEGF-C,-D And VEGFR-3 in primary lung adenocarcinomas (2006)
  • Author(s): Maekawa, S., Iwasaki, A., Enatsu, S., Kinugasa, T., Kuroki, M., et. al.
  • Organizer: 65th Ann. Meet. of Jpn Cancer Assoc.
  • Place of Presentation: Yokohama
  • Year and Date: 2006-09-30
  • Description: 「研究成果報告書概要(欧文)」より
• [Presentation] IgG isotype conversion of a novel anti-carcinoembryonic antigen antibody to increase its biological activity (2006)
  • Author(s): 芝口 浩智, 黒木 求, 衣笠 哲史, 廣瀬 由美子, 山田 博美, ほか
  • Organizer: 第65回日本癌学会学術総会
  • Place of Presentation: 横浜
  • Year and Date: 2006-09-29
  • Description: 「研究成果報告書概要(和文)」より
• [Presentation] IgG isotype conversion of a novel anti-carcinoembryonic antigen antibody to increase its biological activity (2006)
  • Author(s): Shibaguchi, H., Kuroki, M., Kinugasa, T., Hirose, Y., Yamada, H., et. al.
  • Organizer: 65th Ann. Meet. of Jpn Cancer Assoc.
  • Place of Presentation: Yokohama
  • Year and Date: 2006-09-29
  • Description: 「研究成果報告書概要(欧文)」より
• [Book] Recent Developments in Gene Therapy (2007)
  • Author(s): Tanaka, T., Hamada, H., Kuroki, Mo., Kato, K., et. al.
  • Total Pages: 367
  • Publisher: Transworld Research Network
  • Description: 「研究成果報告書概要(和文)」より
• [Book] Recent Developmentsin Gene Therapy(J.Xiang, ed.) (2007)
  • Author(s): Tanaka, T., Hamada, H., Kuroki, Mo., Kato, K., Kinugasa, T., et. al.
  • Total Pages: 367
  • Publisher: Tlransworld Research Network
• [Book] Ransworld Reseaech Network
  • Author(s): Tanaka, T, Hamada, H, Kuroki, M, Kato K.et al.
  • Publisher: Ransworld Research Network(in press)
• [Remarks] 「研究成果報告書概要(和文)」より
  • URL: http://www.med.fukuoka-u.ac.jp/biochem1/MaK-bunken/kenkyugaiyo.html
• [Remarks]
  • URL: http://www.med.fukuoka-u.ac.jp/biochem1/MaK-bunken/kenkyugaiyo.html