In vitro characterization of five humanized OKT3 effector function variant antibodies.
Orthoclone OKT 3 (mOKT3) is a highly effective agent for the reversal of steroid-resistant renal allograft rejection. However, its wider use has been limited by the development of a human anti-mouse antibody response (HAMA) and by the "cytokine release syndrome" (CRS). CRS has been associated with T cell/monocyte activation and, secondarily, with activation of the complement cascade. These processes are mediated through Abs' Fc regions by their abilities to cross-link T cells and mononuclear cells and to activate complements. To alleviate these problems, a group of five huIgG1- and huIgG4-based OKT3 wild-type antibodies and their corresponding Fc mutants with altered residues at amino acids 234, 235, and 318, reported to be required for FcgammaRI and FcgammaRII binding and complement fixation, were constructed. Characterization of these humanized OKT3 Abs, denoted huOKT3gamma1, huOKT3gamma4, huOKT3gamma1(A(234), A(235)), huOKT3gamma4(A(234), A(235)), and huOKT3gamma1(A(318)), has demonstrated that huOKT3gamma1(A(234), A(235)) and huOKT3gamma4(A(234), A(235)), and have at least a 100-fold reduced binding to FcgammaRI and FcgammaRII. As expected, they are much less potent in the induction of T cell activation and cytokine release, yet retain in vitro immunosuppressive effects as potent as those of mOKT3. Unexpectedly, while huOKT3gamma1(A(318)) did not show any reduction in its ability to bind C1q and to fix a complement, huOKT3gamma1(A(234), A(235)) was completely inactive. The in vitro characteristics of huOKT3gamma1(A(234), A(235)) are consistent with recent in vivo studies, in which this Ab showed greatly reduced HAMA and CRS with the retention of its ability to reverse ongoing graft rejection in man.
Xu
D
D
Drug Discovery, R. W. Johnson Pharmaceutical Research Institute, Raritan, New Jersey 08869, USA.
Alegre
M L
ML
Varga
S S
SS
Rothermel
A L
AL
Collins
A M
AM
Pulito
V L
VL
Hanna
L S
LS
Dolan
K P
KP
Parren
P W
PW
Bluestone
J A
JA
Jolliffe
L K
LK
Zivin
R A
RA
D000818:Animals; D000915:Antibody Affinity; D017252:CD3 Complex / Q000276:immunology*; D003167:Complement Activation; D015922:Complement C1q / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D014644:Genetic Variation; D006084:Graft Rejection / Q000188:drug therapy; D006801:Humans; D007127:Immunoglobulin Constant Regions / Q000235:genetics; D007143:Immunoglobulin Heavy Chains / Q000235:genetics; D007135:Immunoglobulin Variable Region / Q000235:genetics; D007166:Immunosuppressive Agents / Q000276:immunology* / Q000302:isolation & purification / Q000494:pharmacology; D016030:Kidney Transplantation / Q000276:immunology; D008213:Lymphocyte Activation; D051379:Mice; D016853:Muromonab-CD3 / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification / Q000494:pharmacology; D016296:Mutagenesis; D011485:Protein Binding; D015202:Protein Engineering / Q000379:methods; D017452:Receptors, IgG / Q000378:metabolism; D011994:Recombinant Proteins / Q000302:isolation & purification; D013601:T-Lymphocytes / Q000276:immunology
D017252:CD3 Complex; D007127:Immunoglobulin Constant Regions; D007143:Immunoglobulin Heavy Chains; D007135:Immunoglobulin Variable Region; D007166:Immunosuppressive Agents; D016853:Muromonab-CD3; D017452:Receptors, IgG; D011994:Recombinant Proteins; D015922:Complement C1q