Cancer Immunology, Immunotherapy

, Volume 53, Issue 6, pp 497–509

A highly effective and stable bispecific diabody for cancer immunotherapy: cure of xenografted tumors by bispecific diabody and T-LAK cells

Authors

  • Hiroki Hayashi
    • Division of Gastroenterological Surgery, Department of SurgeryGraduate School of Medicine, Tohoku University
  • Ryutaro Asano
    • Cell Resource Center for Biomedical ResearchInstitute of Development Aging and Cancer, Tohoku University
  • Kouhei Tsumoto
    • Department of Biochemistry and Engineering, Graduate School of EngineeringTohoku University
  • Yu Katayose
    • Division of Gastroenterological Surgery, Department of SurgeryGraduate School of Medicine, Tohoku University
  • Masanori Suzuki
    • Division of Gastroenterological Surgery, Department of SurgeryGraduate School of Medicine, Tohoku University
  • Michiaki Unno
    • Division of Gastroenterological Surgery, Department of SurgeryGraduate School of Medicine, Tohoku University
  • Hideaki Kodama
    • Division of Gastroenterological Surgery, Department of SurgeryGraduate School of Medicine, Tohoku University
  • Shin-ichi Takemura
    • Division of Gastroenterological Surgery, Department of SurgeryGraduate School of Medicine, Tohoku University
  • Hiroshi Yoshida
    • Division of Gastroenterological Surgery, Department of SurgeryGraduate School of Medicine, Tohoku University
  • Koki Makabe
    • Department of Biochemistry and Engineering, Graduate School of EngineeringTohoku University
  • Kohzoh Imai
    • Department of Internal MedicineSapporo Medical University School of Medicine
  • Seiki Matsuno
    • Division of Gastroenterological Surgery, Department of SurgeryGraduate School of Medicine, Tohoku University
  • Izumi Kumagai
    • Department of Biochemistry and Engineering, Graduate School of EngineeringTohoku University
    • Cell Resource Center for Biomedical ResearchInstitute of Development Aging and Cancer, Tohoku University
Original Article

DOI: 10.1007/s00262-003-0465-9

Cite this article as:
Hayashi, H., Asano, R., Tsumoto, K. et al. Cancer Immunol Immunother (2004) 53: 497. doi:10.1007/s00262-003-0465-9

Abstract

Purpose

In the field of cancer immunotherapy research, the targeting of effector cells with specific antibodies is a very promising approach. Recent advances in genetic engineering have made it possible to prepare immunoglobulin fragments consisting of variable domains using bacterial expression systems.

Methods

We have produced an anti-epidermal growth-factor receptor (EGFR) × anti-CD3 bispecific diabody (Ex3 diabody) in an Escherichia coli (E. coli) expression system with refolding method. The Ex3 diabody targets lymphokine-activated killer cells with a T-cell phenotype (T-LAK cells) to EGFR positive bile duct carcinoma cells with dramatic enhancement of cytotoxicity in vitro. This specific killing of EGFR-positive cells was completely inhibited by parental mAb IgGs directed to EGFR and the CD3 antigen.

Results

When T-LAK cells were cultured with EGFR-positive tumor cells in the presence of Ex3 diabody, they produced much higher levels of IFN-γ, GM-CSF, and TNF-α than in its absence, this being a possible mechanism underlying specific antitumor activity. The Ex3 diabody showed good stability when tested at 37°C for 48 h, and also markedly inhibited tumor growth of bile duct carcinoma xenografts in severe combined immunodeficient (SCID) mice. When Ex3 diabody (20 μg/mouse) was administrated intravenously, together with T-LAK cells and interleukin-2 (IL-2), complete cure of tumors were observed in three of six mice, and the other three showed marked retardation of tumor growth.

Conclusion

The Ex3 diabody can be considered a highly promising reagent for study of specific targeting immunotherapy against bile duct and other EGFR-positive carcinomas.

Keywords

Bispecific diabodyRefolding systemBile duct carcinomaEGF receptorTargeting immunotherapy

Copyright information

© Springer-Verlag 2003