Modulation of antigen-specific T cell response by a non-mitogenic anti-CD3 antibody

Quantitative in vivo comparisons of the Fcγ receptor-dependent agonist activities of different fucosylation variants of an immunoglobulin G antibody

“…2A). These results were consistent with previous data obtained with these Abs [23] and support the notion that T cell activation is dependent on FcγR binding by the anti-CD3 Ab.…”

“…The heavy and light chain variable region coding sequences in this plasmid were previously PCRamplified and the amplified DNA fragments cloned first into genomic heavy and light chain variable region vectors, and then into genomic constant region expression vectors for mouse IgG2a and kappa chains, respectively [23]. At the same time, the heavy chain variable region coding sequence was also cloned into a variant mouse IgG2a expression vector that encoded Ala residues instead of Leu residues at positions 234 and 235 (EU numbering), to prepare a mutant control Ab (2C11 muG2a AlaAla) that lacked FcγR binding.…”

“…At the same time, the heavy chain variable region coding sequence was also cloned into a variant mouse IgG2a expression vector that encoded Ala residues instead of Leu residues at positions 234 and 235 (EU numbering), to prepare a mutant control Ab (2C11 muG2a AlaAla) that lacked FcγR binding. These plasmids were introduced into mouse myeloma cells by transfection, and stable IgG-secreting clonal cell lines used to prepare protein A-purified lots of the IgG2a and IgG2a AlaAla versions of 2C11 (2C11 muG2a and 2C11 muG2a AlaAla, respectively) [23].…”

“…1). The mouse IgG2a and IgG2a AlaAla (two alanine substitutions in the lower hinge region to diminish FcγR binding, [23,29]) variants of 2C11, referred to as 2C11 muG2a and 2C11 muG2a AlaAla, were expressed in stably-transfected mouse Sp2/0 myeloma cells as described previously [23]. Human IgG1 and IgG4 versions of 2C11 were expressed in human HEK 293E cells and both found to be 90% fucosylated, as defined by the percent of Fc glycan structures (2 per Ab) in the population of Ab molecules that contained core fucose.…”

The novel non-mitogenic anti-CD3 antibody, mini-yCD3, delivers a partial TCR signal

“…As a result, several efforts to identify and design nonmitogenic or low-mitogenic anti-CD3 antibodies have been made and also shown to have efficacy in clinical settings [1][2][3][4]. A partial T-cell receptor signal delivered by these non-mitogenic anti-CD3 antibodies is crucial in rendering activated T cells unresponsive [10,[26][27][28].…”