Enhanced Fc-dependent cellular cytotoxicity of Fc fusion proteins derived from TNF receptor II and LFA-3 by fucose removal from Asn-linked oligosaccharides.
Fucose removal from complex-type oligosaccharide of human IgGs results in a major enhancement of Fc-dependent cellular cytotoxicity. The aim of this study was to determine the effect of fucose removal on the effector function of another class of clinically important molecules that can effect cellular cytotoxicity, Fc fusion proteins. The receptors chosen for study were TNF receptor II and LFA-3, both of which have therapeutic significance. The fucosylated versions of these fusion proteins were produced in unmodified CHO cells, whereas the nonfucosylated counterparts were produced in CHO cells with alpha-1,6-fucosyltransferase, an enzyme required for fucosylation, knocked-out. Whilst binding activity of TNFRII-Fc and LFA-3-Fc were unchanged by fucose-removal, nonfucosylated Fc fusion proteins exhibited significantly higher Fc receptor gammaIIIa-binding and increased Fc-mediated cytotoxicity on target cells compared to fucosylated counterparts. Notably, in case of TNFRII-Fc, only the nonfucosylated protein exhibited potent Fc dependent cytotoxicity to transmembrane TNF-alpha expressing cells. These results prove that enhancement of Fc dependent cellular cytotoxicity by fucose-removal is effective in not only whole IgG but also Fc fusion proteins, and thus widens the potential of Fc-fusion proteins as therapeutic candidates.
Shoji-Hosaka
Emi
E
Department of Antibody Research, Pharmaceutical Research Center, Kyowa Hakko Kogyo Co., Ltd., Machida-shi, Tokyo 194-8533.
Kobayashi
Yukari
Y
Wakitani
Masako
M
Uchida
Kazuhisa
K
Niwa
Rinpei
R
Nakamura
Kazuyasu
K
Shitara
Kenya
K
D000077944:Alefacept; D000818:Animals; D000920:Antibody-Dependent Cell Cytotoxicity / Q000187:drug effects; D018968:CD58 Antigens / Q000276:immunology; D016466:CHO Cells; D006224:Cricetinae; D003412:Cricetulus; D000068800:Etanercept; D005643:Fucose / Q000737:chemistry*; D006801:Humans; D007141:Immunoglobulin Fc Fragments / Q000276:immunology*; D007074:Immunoglobulin G / Q000276:immunology*; D009844:Oligosaccharides / Q000737:chemistry / Q000276:immunology*; D017452:Receptors, IgG / Q000276:immunology; D018124:Receptors, Tumor Necrosis Factor / Q000276:immunology*; D011993:Recombinant Fusion Proteins / Q000276:immunology*
D018968:CD58 Antigens; C496124:FCGR3A protein, human; D007141:Immunoglobulin Fc Fragments; D007074:Immunoglobulin G; D009844:Oligosaccharides; D017452:Receptors, IgG; D018124:Receptors, Tumor Necrosis Factor; D011993:Recombinant Fusion Proteins; D005643:Fucose; D000077944:Alefacept; D000068800:Etanercept