Anti-CD3 × anti-GD2 bispecific antibody redirects T-cell cytolytic activity to neuroblastoma targets

Pediatr Blood Cancer. 2012 Dec 15;59(7):1198-205. doi: 10.1002/pbc.24237. Epub 2012 Jun 15.


Background: The ganglioside GD2 is an attractive target for immunotherapy of neuroectodermal tumors. We tested a unique bispecific antibody anti-CD3 × anti-GD2 (3F8BiAb) for its ability to redirect activated T cells (ATC) to target GD2-positive neuroblastomas.

Procedure: ATC were generated from normal human peripheral blood mononuclear cells (PBMC) by stimulating the PBMC with OKT3 and expanding the T cells in the presence of interleukin 2 (IL-2) for 14 days. ATC were armed with 3F8BiAb (100 ng/10(6) cells) or Her2BiAb (50 ng/10(6) cells) prior to use. 3F8 BiAb were tested for its dual-binding specificity to GD2 expressed on cancer cell lines and CD3 expressed on ATC. 3F8BiAb-armed ATC were further tested ex vivo for their cytotoxicity against GD2 positive tumor targets and its ability to induce cytokine response upon binding to targets.

Results: GD2 expression in neuroblastoma cells was confirmed by FACS analysis. Specific binding of 3F8BiAb to the tumor targets as well as to ATC was confirmed by FACS analysis. 3F8BiAb-armed ATC exhibited specific killing of GD2 positive neuroblastoma cell lines significantly above unarmed ATC (P < 0.001). GD2BiAb-armed ATC secreted significantly higher levels of Th(1) cytokines and chemokines compared to unarmed ATC (P < 0.001).

Conclusions: These preclinical findings support the potential of a novel immunotherapeutic approach to target T cells to neuroblastoma.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antibodies, Bispecific / immunology*
  • Antibodies, Bispecific / therapeutic use
  • Binding Sites, Antibody
  • CD3 Complex / immunology*
  • Cell Line, Tumor
  • Cytokines / metabolism
  • Cytotoxicity Tests, Immunologic
  • Gangliosides / immunology*
  • Gangliosides / metabolism
  • Humans
  • Immunotherapy
  • Lymphocyte Activation
  • Neuroblastoma / immunology*
  • Neuroblastoma / metabolism
  • Neuroblastoma / therapy
  • Receptor, ErbB-2 / immunology
  • Receptor, ErbB-2 / metabolism
  • T-Lymphocytes, Cytotoxic / immunology*


  • Antibodies, Bispecific
  • CD3 Complex
  • Cytokines
  • Gangliosides
  • ganglioside, GD2
  • Receptor, ErbB-2