Human T cells resistant to complement lysis by bivalent antibody can be efficiently lysed by dimers of monovalent antibody.
We have previously shown that bivalent human gamma1 CD3 monoclonal antibody (mAb) is ineffective at mediating lysis of human T cells with human complement. In this paper we have used genetic engineering and sulfur chemistry to prepare 2 types of human gamma1 CD3 mAb dimer, with the aim of improving complement lysis activity. The IgG molecules forming the dimers were linked together at their C-termini by stable bismaleimide thioether bridges. The first dimer was composed of 2 bivalent mAb molecules. This dimer proved incapable of lysing human T-cell blasts with human, rabbit, or guinea pig complement. The second dimer consisted of 2 molecules of a monovalent derivative (possessing a single Fab domain) of the bivalent mAb. This dimer was highly lytic with human complement, with a lytic titer 64-fold greater than that of the nondimerized monovalent mAb. The maximum level of lysis of human T-cell blasts achieved with this monovalent mAb dimer was equal to that obtained with the therapeutic antilymphocyte mAb alemtuzumab, but its lytic titer was 4-fold greater. The monovalent mAb dimer was also found to be lytic in the presence of rabbit and guinea pig complement. Dimerization of monovalent antibodies may provide a general strategy for improving the cytolytic activity of other mAbs that are normally unable to induce lysis with complement. The monovalent CD3 mAb dimer may have potential for development as an agent for immunotherapy of T-cell leukemia.
Nielsen
Soren U
SU
Department of Microbiology and Immunology, University of Newcastle upon Tyne, Medical School, England. s.u.nielsen@ncl.ac.uk
Routledge
Edward G
EG
D000911:Antibodies, Monoclonal / Q000737:chemistry / Q000276:immunology* / Q000302:isolation & purification; D000915:Antibody Affinity; D017252:CD3 Complex / Q000276:immunology*; D003165:Complement System Proteins / Q000276:immunology; D003432:Cross-Linking Reagents; D003601:Cytotoxicity Tests, Immunologic; D003602:Cytotoxicity, Immunologic; D019281:Dimerization; D006801:Humans; D007074:Immunoglobulin G; D008301:Maleimides; D013601:T-Lymphocytes / Q000276:immunology*
D000911:Antibodies, Monoclonal; D017252:CD3 Complex; D003432:Cross-Linking Reagents; D007074:Immunoglobulin G; D008301:Maleimides; C043592:maleimide; D003165:Complement System Proteins