Human neutrophil interactions of a bispecific monoclonal antibody targeting tumor and human Fcγ RIII

Abstract

2B1 is a bispecific murine monoclonal antibody (bsmAb) targeting the c-erbB-2 and CD16 (Fc gamma RIII) antigens. c-erbB-2 is over-expressed by a variety of adenocarcinomas, and CD16, the low-affinity Fc gamma receptor for aggregated immunoglobulins, is expressed by polymorphonuclear leukocytes (PMN), natural killer (NK) cells and differentiated mononuclear phagocytes. 2B1 potentiates the in vitro lysis of c-erb-2 over-expressing tumors by NK cells and macrophages. In this report, the interactions between 2B1 and PMN were investigated to assess the impact of these associations on in vitro 2B1-promoted tumor cytotoxicity by human NK cells. The peak binding of 2B1 to PMN was observed at a concentration of 10 microgram/ml 2B1. However, 2B1 rapidly dissociated from PMN in vitro at 37 degrees C in non-equilibrium conditions. This dissociation was not caused by CD16 shedding. When PMN were labeled witn 125I-2B1 and incubated at 37 degrees C and the supernatants examined by HPLC analysis, the Fab regions of dissociated 2B1 were not complexed with shed CD16 extracellular domain. While most of the binding of 2B1 PMN was solely attributable to Fab-directed binding to Fc gamma RIII, PMN-associated 2B1 also bound through Fc gamma-domain/Fc gamma RII interactions. 2B1 did not promote in vitro PMN cytotoxicity against c-erbB-2-expressing SK-OV-3 tumor cells. When PMN were coincubated with peripheral blood lymphocytes, SK-OV-3 tumor and 2B1, the concentration of 2B1 required for maximal tumor lysis was lowered. Although PMN may serve as a significant competitive binding pool of systemically administered 2B1 in vivo, the therapeutic potential of the targeted cytotoxicity properties of this bsmAb should not be compromised.

Bypassing tumor-specific and bispecific antibodies: triggering of antitumor immunity by expression of anti-FcγR scFv on cancer cell surface

“…29 Our data also show that it is possible to block the negative immuno-modulatory function exerted by KIR on NK cells by expressing anti-CD16 antibodies on HLA + tumor cells otherwise resistant or poorly sensitive to NK cell activity. The balance between the positive signal triggered by the engagement of FcγRIII-A on NK cells and the negative signal due to the KIR/HLA interaction can therefore be modified in favor of the former one by expressing anti-FcγR scFv at the tumor surface.…”

Bispecific Minibodies Targeting HER2/neu and CD16 Exhibit Improved Tumor Lysis When Placed in a Divalent Tumor Antigen Binding Format

“…Accordingly, the antibody cross-linked neutrophils and mononuclear phagocytes, leading to cytokine release (41,43). To address this problem we created recombinant single-chain Fv-based bispecific antibodies that bind to tumor antigens with high affinity (44 -46) and to CD16 with a lower affinity.…”

New approaches to antibody therapy

“…Some tumor in®ltration by leukocytes has been seen following MDX-210 therapy (Link et al, 1998), and we have observed the induction of tumor in®ltration by leukocytes in several patients treated with 2B1. However, there is no consistent, profound leukocyte in®ltration of tumor following BsAb therapy, and preclinical data suggest that BsAb promoted cytotoxicity will require eector to target ratios of at least 1 : 1 for successful therapy (Weiner et al, 1993b). It is probable that eective BsAb treatment strategies must include ways to induce tumor in¯ammation so that BsAb-mediated potentiation of cytotoxicity can be properly exploited.…”