Chimeric Anti-Human Podoplanin Antibody NZ-12 of Lambda Light Chain Exerts Higher Antibody-Dependent Cellular Cytotoxicity and Complement-Dependent Cytotoxicity Compared with NZ-8 of Kappa Light Chain.

Abstract

Podoplanin (PDPN), a type I transmembrane 36-kDa glycoprotein, is expressed not only in normal cells, such as renal epithelial cells (podocytes), lymphatic endothelial cells, and pulmonary type I alveolar cells, but also in cancer cells, including brain tumors and lung squamous cell carcinomas. Podoplanin activates platelet aggregation by binding to C-type lectin-like receptor-2 (CLEC-2) on platelets, and the podoplanin/CLEC-2 interaction facilitates blood/lymphatic vessel separation. We previously produced neutralizing anti-human podoplanin monoclonal antibody (mAb), clone NZ-1 (rat IgG2a, lambda), which neutralizes the podoplanin/CLEC-2 interaction and inhibits platelet aggregation and cancer metastasis. Human-rat chimeric antibody, NZ-8, was previously developed using variable regions of NZ-1 and human constant regions of heavy chain (IgG1) and light chain (kappa chain). Although NZ-8 showed high antibody-dependent cellular cytotoxicity (ADCC) and complement-dependent cytotoxicity (CDC) against human podoplanin-expressing cancer cells, the binding affinity of NZ-8 was lower than that of NZ-1. Herein, we produced a novel human-rat chimeric antibody, NZ-12, the constant regions of which consist of IgG1 heavy chain and lambda light chain. Using flow cytometry, we demonstrated that the binding affinity of NZ-12 was much higher than that of NZ-8. Furthermore, ADCC and CDC activities of NZ-12 were significantly increased against glioblastoma cell lines (LN319 and D397) and lung cancer cell line (PC-10). These results suggested that NZ-12 could become a promising therapeutic antibody against podoplanin-expressing brain tumors and lung cancers.

• Authors: Mika K Kaneko, Shinji Abe, Satoshi Ogasawara, Yuki Fujii, Shinji Yamada, Takeshi Murata, Hiroaki Uchida, Hideaki Tahara, Yasuhiko Nishioka, Yukinari Kato
• Journal: Monoclonal antibodies in immunodiagnosis and immunotherapy (Monoclon Antib Immunodiagn Immunother) Vol. 36 Issue 1 Pg. 25-29 (Feb 2017) ISSN: 2167-9436 [Electronic] United States
• PMID: 28157429 (Publication Type: Journal Article)

Chemical References

• Antibodies, Anti-Idiotypic
• Antibodies, Monoclonal
• Antigens, Surface
• CLEC2B protein, human
• Immunoglobulin kappa-Chains
• Immunoglobulin lambda-Chains
• Lectins, C-Type
• Membrane Glycoproteins
• PDPN protein, human
• Platelet Aggregation Inhibitors
• Complement System Proteins

Topics

• Animals
• Antibodies, Anti-Idiotypic (immunology)
• Antibodies, Monoclonal (immunology)
• Antibody Affinity (immunology)
• Antibody-Dependent Cell Cytotoxicity (immunology)
• Antigens, Surface (immunology)
• CHO Cells
• Cell Line, Tumor
• Complement System Proteins (immunology)
• Cricetulus
• Glioblastoma (immunology)
• Humans
• Immunoglobulin kappa-Chains (immunology)
• Immunoglobulin lambda-Chains (immunology)
• Lectins, C-Type (immunology, metabolism)
• Lung Neoplasms (immunology)
• Membrane Glycoproteins (immunology, metabolism)
• Platelet Aggregation (immunology)
• Platelet Aggregation Inhibitors (immunology, pharmacology)
• Protein Binding
• Rats