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.
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 IgG1heavy 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.
1 Tohoku University Graduate School of Medicine , Sendai, Japan .
Mika K
MK
Kaneko
2 Department of Clinical Pharmacy Practice Pedagogy, Graduate School of Biomedical Sciences, Tokushima University , Tokushima, Japan .|3 Department of Respiratory Medicine and Rheumatology, Graduate School of Biomedical Sciences, Tokushima University , Tokushima, Japan .
Shinji
S
Abe
1 Tohoku University Graduate School of Medicine , Sendai, Japan .|4 Department of Chemistry, Graduate School of Science, Chiba University , Chiba, Japan .|5 Molecular Chirality Research Center, Chiba University , Chiba, Japan .
Satoshi
S
Ogasawara
1 Tohoku University Graduate School of Medicine , Sendai, Japan .
Yuki
Y
Fujii
1 Tohoku University Graduate School of Medicine , Sendai, Japan .
Shinji
S
Yamada
4 Department of Chemistry, Graduate School of Science, Chiba University , Chiba, Japan .|5 Molecular Chirality Research Center, Chiba University , Chiba, Japan .
Takeshi
T
Murata
6 Division of Bioengineering, Advanced Clinical Research Center, The Institute of Medical Science, The University of Tokyo , Tokyo, Japan .
Hiroaki
H
Uchida
6 Division of Bioengineering, Advanced Clinical Research Center, The Institute of Medical Science, The University of Tokyo , Tokyo, Japan .
Hideaki
H
Tahara
3 Department of Respiratory Medicine and Rheumatology, Graduate School of Biomedical Sciences, Tokushima University , Tokushima, Japan .
Yasuhiko
Y
Nishioka
1 Tohoku University Graduate School of Medicine , Sendai, Japan .
Yukinari
Y
Kato
D000818:Animals; D000888:Antibodies, Anti-Idiotypic / Q000276:immunology; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000915:Antibody Affinity / Q000276:immunology; D000920:Antibody-Dependent Cell Cytotoxicity / Q000276:immunology*; D000954:Antigens, Surface / Q000276:immunology; D016466:CHO Cells; D045744:Cell Line, Tumor; D003165:Complement System Proteins / Q000276:immunology*; D003412:Cricetulus; D005909:Glioblastoma / Q000276:immunology*; D006801:Humans; D007145:Immunoglobulin kappa-Chains / Q000276:immunology*; D007146:Immunoglobulin lambda-Chains / Q000276:immunology*; D037181:Lectins, C-Type / Q000276:immunology / Q000378:metabolism; D008175:Lung Neoplasms / Q000276:immunology*; D008562:Membrane Glycoproteins / Q000276:immunology* / Q000378:metabolism; D010974:Platelet Aggregation / Q000276:immunology; D010975:Platelet Aggregation Inhibitors / Q000276:immunology / Q000494:pharmacology; D011485:Protein Binding; D051381:Rats; D000098853:Podoplanin
D000888:Antibodies, Anti-Idiotypic; D000911:Antibodies, Monoclonal; D000954:Antigens, Surface; D003165:Complement System Proteins; D007145:Immunoglobulin kappa-Chains; D007146:Immunoglobulin lambda-Chains; D037181:Lectins, C-Type; D008562:Membrane Glycoproteins; D010975:Platelet Aggregation Inhibitors; C104778:CLEC2B protein, human; C117468:PDPN protein, human; D000098853:Podoplanin
chimeric antibody; human podoplanin; kappa chain; lambda chain; monoclonal antibody