Negative interference with antibody-dependent cellular cytotoxicity mediated by rituximab from its interactions with human serum proteins

Saeko Yanaka; Rina Yogo; Hirokazu Yagi; Masayoshi Onitsuka; Natsumi Wakaizumi; Yuki Yamaguchi; Susumu Uchiyama; Koichi Kato

doi:10.3389/fimmu.2023.1090898

Negative interference with antibody-dependent cellular cytotoxicity mediated by rituximab from its interactions with human serum proteins

Front Immunol. 2023 Jan 25:14:1090898. doi: 10.3389/fimmu.2023.1090898. eCollection 2023.

Authors

Saeko Yanaka^{1

2

3

4}, Rina Yogo^{1

2

3}, Hirokazu Yagi^{1

3}, Masayoshi Onitsuka⁵, Natsumi Wakaizumi⁶, Yuki Yamaguchi⁶, Susumu Uchiyama⁶, Koichi Kato^{1

2

3}

Affiliations

¹ Exploratory Research Center on Life and Living Systems (ExCELLS), National Institutes of Natural Sciences, Okazaki, Japan.
² Institute for Molecular Science (IMS), National Institutes of Natural Sciences, Okazaki, Japan.
³ Graduate School of Pharmaceutical Sciences, Nagoya City University, Nagoya, Japan.
⁴ Graduate School of Pharmaceutical Sciences, Kyushu University, Fukuoka, Japan.
⁵ Graduate School of Technology, Industrial and Social Sciences, Tokushima University, Tokushima, Japan.
⁶ Graduate School of Engineering, University of Osaka, Osaka, Japan.

Abstract

Although interactions of small molecular drugs with serum proteins have been widely studied from pharmacokinetic and pharmacodynamic perspectives, there have been few reports on the effects of serum components on therapeutic antibody functions. This study reports the effect of abundant serum proteins on antibody-dependent cellular cytotoxicity (ADCC) mediated by rituximab and Fcγ receptor III (FcγRIII). Human serum albumin (HSA) and the Fab fragment from the pooled serum polyclonal IgG were found to compromise ADCC as non-competitive inhibitors. Our nuclear magnetic resonance data provided direct evidence for the interactions of HSA with both the Fab and Fc regions of rituximab and also with the extracellular region of FcγRIII (sFcγRIII). The degree of involvement in the interaction decreased in the order of rituximab-Fab > rituximab-Fc > sFcγRIII, suggesting preferential binding of HSA to net positively charged proteins. Although much less pronounced than the effect of HSA, polyclonal IgG-Fab specifically interacted with rituximab-Fc. The NMR data also showed that the serum protein interactions cover the Fc surface extensively, suggesting that they can act as pan-inhibitors against various Fc receptor-mediated functions and pharmacokinetics. Our findings highlight the importance of considering serum-protein interactions in the design and application of antibody-based drugs with increased efficacy and safety.

Keywords: NMR; antibody-dependent cellular cytotoxicity; human serum albumin; stable isotope labeling; therapeutic antibody.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Antibody-Dependent Cell Cytotoxicity*
Humans
Immunoglobulin G*
Phagocytosis
Receptors, Fc
Rituximab

Substances

Rituximab
Immunoglobulin G
Receptors, Fc

Grants and funding

This study was supported in part by the Nanotechnology Platform Program (Molecule and Material Synthesis) and by the Advanced NMR Application and Platform (program No. PF17-01-R-008 and PF19-01-R-022) of the Ministry of Education, Culture, Sports, Science and Technology (MEXT), and by Joint Research of the Exploratory Research Center on Life and Living Systems (ExCELLS) (ExCELLS program No.20-308 and 21-308 to HY). This study was also supported by Japan and MEXT/JSPS Grants-in-Aid for Scientific Research (Grant Numbers, JP19J15602 to RY; JP17H05893, JP20K15981, and JP22H02755 to SY; and JP19H01017 to KK) as well as AMED (Grant Numbers JP21ae0121020h0001 to SY and JP21ae0121013h0301 to KK).