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

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) x 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-gamma, GM-CSF, and TNF-alpha than in its absence, this being a possible mechanism underlying specific antitumor activity. The Ex3 diabody showed good stability when tested at 37 degrees 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 microg/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.

• Authors: Hiroki Hayashi, Ryutaro Asano, Kouhei Tsumoto, Yu Katayose, Masanori Suzuki, Michiaki Unno, Hideaki Kodama, Shin-ichi Takemura, Hiroshi Yoshida, Koki Makabe, Kohzoh Imai, Seiki Matsuno, Izumi Kumagai, Toshio Kudo
• Journal: Cancer immunology, immunotherapy : CII (Cancer Immunol Immunother) Vol. 53 Issue 6 Pg. 497-509 (Jun 2004) ISSN: 0340-7004 [Print] Germany
• PMID: 14648071 (Publication Type: Comparative Study, Journal Article)

Chemical References

• Antibodies, Bispecific
• CD3 Complex
• Immunoglobulin G
• Interleukin-2
• Recombinant Fusion Proteins
• Tumor Necrosis Factor-alpha
• Interferon-gamma
• Granulocyte-Macrophage Colony-Stimulating Factor
• ErbB Receptors

Topics

• Animals
• Antibodies, Bispecific (immunology, therapeutic use)
• Bile Duct Neoplasms (immunology, therapy)
• CD3 Complex (immunology)
• Cells, Cultured
• ErbB Receptors (immunology)
• Escherichia coli (metabolism)
• Female
• Genetic Vectors (genetics)
• Granulocyte-Macrophage Colony-Stimulating Factor (metabolism)
• Humans
• Hybridomas (immunology)
• Immunoglobulin G (immunology)
• Immunotherapy
• Interferon-gamma (metabolism)
• Interleukin-2 (metabolism)
• Killer Cells, Lymphokine-Activated (immunology)
• Mice
• Mice, Inbred ICR
• Mice, SCID
• Phenotype
• Recombinant Fusion Proteins (immunology, therapeutic use)
• T-Lymphocytes (immunology)
• Tumor Necrosis Factor-alpha (metabolism)
• Xenograft Model Antitumor Assays