Afucosylated antibodies increase activation of FcγRIIIa-dependent signaling components to intensify processes promoting ADCC.
Antibody-dependent cellular cytotoxicity (ADCC) is a key mechanism by which therapeutic antibodies mediate their antitumor effects. The absence of fucose on the heavy chain of the antibody increases the affinity between the antibody and FcγRIIIa, which results in increased in vitro and in vivo ADCC compared with the fucosylated form. However, the cellular and molecular mechanisms responsible for increased ADCC are unknown. Through a series of biochemical and cellular studies, we find that human natural killer (NK) cells stimulated with afucosylated antibody exhibit enhanced activation of proximal FcγRIIIa signaling and downstream pathways, as well as enhanced cytoskeletal rearrangement and degranulation, relative to stimulation with fucosylated antibody. Furthermore, analysis of the interaction between human NK cells and targets using a high-throughput microscope-based antibody-dependent cytotoxicity assay shows that afucosylated antibodies increase the number of NK cells capable of killing multiple targets and the rate with which targets are killed. We conclude that the increase in affinity between afucosylated antibodies and FcγRIIIa enhances activation of signaling molecules, promoting cytoskeletal rearrangement and degranulation, which, in turn, potentiates the cytotoxic characteristics of NK cells to increase efficiency of ADCC.
Department of Pathology, Genentech, Inc., South San Francisco, California. liu.scot@gene.com.
Scot D
SD
Liu
Department of Pathology, Genentech, Inc., South San Francisco, California.
Cecile
C
Chalouni
Department of Biochemical and Cellular Pharmacology, Genentech, Inc., South San Francisco, California.
Judy C
JC
Young
Department of Cancer Immunotherapy and Hematology, Genentech, Inc., South San Francisco, California.
Teemu T
TT
Junttila
Department of Molecular Oncology, Genentech, Inc., South San Francisco, California.
Mark X
MX
Sliwkowski
Department of Pathology, Genentech, Inc., South San Francisco, California.
John B
JB
Lowe
D000199:Actins / Q000378:metabolism; D061067:Antibodies, Monoclonal, Humanized / Q000737:chemistry / Q000276:immunology*; D000915:Antibody Affinity / Q000276:immunology; D000920:Antibody-Dependent Cell Cytotoxicity / Q000276:immunology*; D015550:Cell Degranulation / Q000276:immunology; D002478:Cells, Cultured; D003599:Cytoskeleton / Q000378:metabolism; D005643:Fucose / Q000276:immunology*; D006801:Humans; D007694:Killer Cells, Natural / Q000276:immunology*; D017452:Receptors, IgG / Q000276:immunology*; D015398:Signal Transduction / Q000276:immunology; D000068878:Trastuzumab / Q000737:chemistry / Q000276:immunology
D000199:Actins; D061067:Antibodies, Monoclonal, Humanized; C496124:FCGR3A protein, human; D017452:Receptors, IgG; D005643:Fucose; C543332:obinutuzumab; D000068878:Trastuzumab