title: Two mechanisms of the enhanced antibody-dependent cellular cytotoxicity (ADCC) efficacy of non-fucosylated therapeutic antibodies in human blood.
abstract: BACKGROUND
  Antibody-dependent cellular cytotoxicity (ADCC) has recently been identified as one of the critical mechanisms underlying the clinical efficacy of therapeutic antibodies, especially anticancer antibodies. Therapeutic antibodies fully lacking the core fucose of the Fc oligosaccharides have been found to exhibit much higher ADCC in humans than their fucosylated counterparts. However, data which show how fully non-fucosylated antibodies achieve such a high ADCC in human whole blood have not yet been disclosed. The precise mechanisms responsible for the high ADCC mediated by fully non-fucosylated therapeutic antibodies, even in the presence of human plasma, should be explained based on direct evidence of non-fucosylated antibody action in human blood.
  
  
  METHODS
  Using a human ex vivo B-cell depletion assay with non-fucosylated and fucosylated anti-CD20 IgG1s rituximab, we monitored the binding of the therapeutic agents both to antigens on target cells (target side interaction) and to leukocyte receptors (FcgammaR) on effector cells (effector side interaction), comparing the intensities of ADCC in human blood.
  
  
  RESULTS
  In the target side interaction, down-modulation of CD20 on B cells mediated by anti-CD20 was not observed. Simple competition for binding to the antigens on target B cells between fucosylated and non-fucosylated anti-CD20s was detected in human blood to cause inhibition of the enhanced ADCC of non-fucosylated anti-CD20 by fucosylated anti-CD20. In the effector side interaction, non-fucosylated anti-CD20 showed sufficiently high FcgammaRIIIa binding activity to overcome competition from plasma IgG for binding to FcgammaRIIIa on natural killer (NK) cells, whereas the binding of fucosylated anti-CD20 to FcgammaRIIIa was almost abolished in the presence of human plasma and failed to recruit NK cells effectively. The core fucosylation levels of individual serum IgG1 from healthy donors was found to be so slightly different that it did not affect the inhibitory effect on the ADCC of fucosylated anti-CD20.
  
  
  CONCLUSIONS
  Our results demonstrate that removal of fucosylated antibody ingredients from antibody therapeutics elicits high ADCC in human blood by two mechanisms: namely, by evading the inhibitory effects both of plasma IgG on FcgammaRIIIa binding (effector side interaction) and of fucosylated antibodies on antigen binding (target side interaction).
authors.affiliation: Tokyo Research Laboratories, Kyowa Hakko Kogyo Co, Ltd, 3-6-6 Asahi-machi, Machida-shi, Tokyo, Japan. shigeru.iida@kyowa-kirin.co.jp
authors.forename: Shigeru
authors.initials: S
authors.lastname: Iida
authors.forename: Reiko
authors.initials: R
authors.lastname: Kuni-Kamochi
authors.forename: Katsuhiro
authors.initials: K
authors.lastname: Mori
authors.forename: Hirofumi
authors.initials: H
authors.lastname: Misaka
authors.forename: Miho
authors.initials: M
authors.lastname: Inoue
authors.forename: Akira
authors.initials: A
authors.lastname: Okazaki
authors.forename: Kenya
authors.initials: K
authors.lastname: Shitara
authors.forename: Mitsuo
authors.initials: M
authors.lastname: Satoh
mesh_terms: D000328:Adult; D000906:Antibodies / Q000235:genetics / Q000276:immunology / Q000627:therapeutic use*; D000920:Antibody-Dependent Cell Cytotoxicity; D000937:Antigen-Antibody Reactions; D018951:Antigens, CD20 / Q000235:genetics / Q000276:immunology; D001402:B-Lymphocytes / Q000276:immunology; D001769:Blood / Q000276:immunology*; D002478:Cells, Cultured; D005260:Female; D005643:Fucose / Q000276:immunology*; D006801:Humans; D007074:Immunoglobulin G / Q000235:genetics / Q000276:immunology / Q000627:therapeutic use; D007694:Killer Cells, Natural / Q000276:immunology; D007963:Leukocytes, Mononuclear / Q000276:immunology; D008297:Male; D008875:Middle Aged; D055815:Young Adult
chemical_list: D000906:Antibodies; D018951:Antigens, CD20; D007074:Immunoglobulin G; D005643:Fucose