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Establishment of P38Bf, a Core-Fucose-Deficient Mouse-Canine Chimeric Antibody Against Dog Podoplanin

Podoplanin (PDPN), a type I transmembrane sialoglycoprotein, is expressed in normal tissues, including lymphatic endothelial cells, pulmonary type I alveolar cells, and renal podocytes. The overexpression of PDPN in cancers is associated with hematogenous metastasis by interactions with the C-type l...

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Autores principales: Kato, Yukinari, Mizuno, Takuya, Yamada, Shinji, Nakamura, Takuro, Itai, Shunsuke, Yanaka, Miyuki, Sano, Masato, Kaneko, Mika K.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Mary Ann Liebert, Inc., publishers 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6208159/
https://www.ncbi.nlm.nih.gov/pubmed/30362926
http://dx.doi.org/10.1089/mab.2018.0035
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author Kato, Yukinari
Mizuno, Takuya
Yamada, Shinji
Nakamura, Takuro
Itai, Shunsuke
Yanaka, Miyuki
Sano, Masato
Kaneko, Mika K.
author_facet Kato, Yukinari
Mizuno, Takuya
Yamada, Shinji
Nakamura, Takuro
Itai, Shunsuke
Yanaka, Miyuki
Sano, Masato
Kaneko, Mika K.
author_sort Kato, Yukinari
collection PubMed
description Podoplanin (PDPN), a type I transmembrane sialoglycoprotein, is expressed in normal tissues, including lymphatic endothelial cells, pulmonary type I alveolar cells, and renal podocytes. The overexpression of PDPN in cancers is associated with hematogenous metastasis by interactions with the C-type lectin-like receptor 2 (CLEC-2). We have previously reported the development of a mouse monoclonal antibody (mAb) clone, PMab-38 (IgG(1), kappa), against dog PDPN (dPDPN). PMab-38 reacted strongly with canine squamous cell carcinomas and melanomas, but not with lymphatic endothelial cells, indicating its cancer specificity. In this study, we developed and produced several mouse-canine chimeric antibodies originating from PMab-38. A mouse-canine chimeric antibody of subclass A (P38A) and a mouse-canine chimeric antibody of subclass B (P38B) were transiently produced using ExpiCHO-S cells. Core-fucose-deficient P38B (P38Bf) was developed using FUT8 knockout ExpiCHO-S cells. We compared the binding affinities, antibody-dependent cellular cytotoxicity (ADCC), and complement-dependent cytotoxicity (CDC) of P38A, P38B, and P38Bf against Chinese hamster ovary (CHO)/dPDPN cells. Flow cytometry analysis showed that the K(D) of P38A, P38B, and P38Bf were 1.9 × 10(−7), 5.2 × 10(−9), and 6.5 × 10(−9), respectively. Both P38B and P38Bf revealed high ADCC activities against CHO/dPDPN cells; P38Bf demonstrated significantly higher ADCC compared with P38B, especially at low concentrations. P38B and P38Bf exhibited higher CDC activities against CHO/dPDPN cells. Conversely, P38A did not exhibit any ADCC or CDC activity. In summary, P38Bf is a good candidate for antibody therapy against dPDPN-expressing canine cancers.
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spelling pubmed-62081592018-10-31 Establishment of P38Bf, a Core-Fucose-Deficient Mouse-Canine Chimeric Antibody Against Dog Podoplanin Kato, Yukinari Mizuno, Takuya Yamada, Shinji Nakamura, Takuro Itai, Shunsuke Yanaka, Miyuki Sano, Masato Kaneko, Mika K. Monoclon Antib Immunodiagn Immunother Original Articles Podoplanin (PDPN), a type I transmembrane sialoglycoprotein, is expressed in normal tissues, including lymphatic endothelial cells, pulmonary type I alveolar cells, and renal podocytes. The overexpression of PDPN in cancers is associated with hematogenous metastasis by interactions with the C-type lectin-like receptor 2 (CLEC-2). We have previously reported the development of a mouse monoclonal antibody (mAb) clone, PMab-38 (IgG(1), kappa), against dog PDPN (dPDPN). PMab-38 reacted strongly with canine squamous cell carcinomas and melanomas, but not with lymphatic endothelial cells, indicating its cancer specificity. In this study, we developed and produced several mouse-canine chimeric antibodies originating from PMab-38. A mouse-canine chimeric antibody of subclass A (P38A) and a mouse-canine chimeric antibody of subclass B (P38B) were transiently produced using ExpiCHO-S cells. Core-fucose-deficient P38B (P38Bf) was developed using FUT8 knockout ExpiCHO-S cells. We compared the binding affinities, antibody-dependent cellular cytotoxicity (ADCC), and complement-dependent cytotoxicity (CDC) of P38A, P38B, and P38Bf against Chinese hamster ovary (CHO)/dPDPN cells. Flow cytometry analysis showed that the K(D) of P38A, P38B, and P38Bf were 1.9 × 10(−7), 5.2 × 10(−9), and 6.5 × 10(−9), respectively. Both P38B and P38Bf revealed high ADCC activities against CHO/dPDPN cells; P38Bf demonstrated significantly higher ADCC compared with P38B, especially at low concentrations. P38B and P38Bf exhibited higher CDC activities against CHO/dPDPN cells. Conversely, P38A did not exhibit any ADCC or CDC activity. In summary, P38Bf is a good candidate for antibody therapy against dPDPN-expressing canine cancers. Mary Ann Liebert, Inc., publishers 2018-10-01 2018-10-24 /pmc/articles/PMC6208159/ /pubmed/30362926 http://dx.doi.org/10.1089/mab.2018.0035 Text en © Yukinari Kato et al. 2018; Published by Mary Ann Liebert, Inc. This Open Access article is distributed under the terms of the Creative Commons License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Original Articles
Kato, Yukinari
Mizuno, Takuya
Yamada, Shinji
Nakamura, Takuro
Itai, Shunsuke
Yanaka, Miyuki
Sano, Masato
Kaneko, Mika K.
Establishment of P38Bf, a Core-Fucose-Deficient Mouse-Canine Chimeric Antibody Against Dog Podoplanin
title Establishment of P38Bf, a Core-Fucose-Deficient Mouse-Canine Chimeric Antibody Against Dog Podoplanin
title_full Establishment of P38Bf, a Core-Fucose-Deficient Mouse-Canine Chimeric Antibody Against Dog Podoplanin
title_fullStr Establishment of P38Bf, a Core-Fucose-Deficient Mouse-Canine Chimeric Antibody Against Dog Podoplanin
title_full_unstemmed Establishment of P38Bf, a Core-Fucose-Deficient Mouse-Canine Chimeric Antibody Against Dog Podoplanin
title_short Establishment of P38Bf, a Core-Fucose-Deficient Mouse-Canine Chimeric Antibody Against Dog Podoplanin
title_sort establishment of p38bf, a core-fucose-deficient mouse-canine chimeric antibody against dog podoplanin
topic Original Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6208159/
https://www.ncbi.nlm.nih.gov/pubmed/30362926
http://dx.doi.org/10.1089/mab.2018.0035
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