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

Department of Oncology, Wayne State University, Barbara Ann Karmanos Cancer Institute, Detroit, Michigan
Pediatric Blood & Cancer (Impact Factor: 2.39). 12/2012; 59(7):1198-205. DOI: 10.1002/pbc.24237
Source: PubMed


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.
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.
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).
These preclinical findings support the potential of a novel immunotherapeutic approach to target T cells to neuroblastoma. Pediatr Blood Cancer 2012; 59: 1198-1205. © 2012 Wiley Periodicals, Inc.

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Available from: Maxim Yankelevich, Mar 19, 2014
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    • "Previously we have shown that T cells expanded from the leukapheresis product or PBMC from the immunosuppressive environment of cancer patients can be stimulated to improve their functions in vitro as well as in vivo[30,31]. ATC armed with anti-CD3 x anti-tumor associated antigen BiAbs exhibit high levels of specific cytotoxicity against tumor cells expressing Her2/neu [17,18,32], CD20 [15], GD2 [33], and EGFR [16,19] via redirected non-MHC-restricted perforin/granzyme-dependent killing. Furthermore, arming with BiAbs creates artificial cytotoxic T lymphocytes wherein TCR engagement via BiAb-bridge between the armed ATC and the antigen on the tumor cells induces release of significantly higher levels of IFN-γ, IL-2R, IL-12, CCL3, CCL4 and CXCL9, compared to unarmed ATC used as controls [19]. "
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    ABSTRACT: Background Ipilimumab is an antagonistic monoclonal antibody against cytotoxic T-lymphocyte antigen-4 (CTLA-4) that enhances antitumor immunity by inhibiting immunosuppressive activity of regulatory T cells (Treg). In this study, we investigated whether inhibiting Treg activity with ipilimumab during ex vivo T cell expansion could augment anti-CD3-driven T cell proliferation and enhance bispecific antibody (BiAb)-redirected antitumor cytotoxicity of activated T cells (ATC). Methods PBMC from healthy individuals were stimulated with anti-CD3 monoclonal antibody with or without ipilimumab and expanded for 10-14 days. ATC were harvested and armed with anti-CD3 x anti-EGFR BiAb (EGFRBi) or anti-CD3 x anti-CD20 BiAb (CD20Bi) to test for redirected cytotoxicity against COLO356/FG pancreatic cancer cell line or Burkitt’s lymphoma cell line (Daudi). Results In PBMC from healthy individuals, the addition of ipilimumab at the initiation of culture significantly enhanced T cell proliferation (p = 0.0029). ATC grown in the presence of ipilimumab showed significantly increased mean tumor-specific cytotoxicity at effector:target (E:T) ratio of 25:1 directed at COLO356/FG and Daudi by 37.71% (p < 0.0004) and 27.5% (p < 0.0004), respectively, and increased the secretion of chemokines (CCL2, CCL3, CCL4,CCL5, CXCL9, and granulocyte-macrophage colony stimulating factor(GM-CSF)) and cytokines (IFN-γ, IL-2R, IL-12, and IL-13), while reducing IL-10 secretion. Conclusions Expansion of ATC in the presence of ipilimumab significantly improves not only the T cell proliferation but it also enhances cytokine secretion and the specific cytotoxicity of T cells armed with bispecific antibodies.
    Full-text · Article · Jul 2014 · Journal of Translational Medicine
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    • "Arming ATC with HER2Bi or EGFRBi converts every ATC into a specific cytotoxic T cell [3-7]. Our preclinical studies show that armed ATC can target breast [6], prostate [8], ovarian [5] EGFR+ cancers (head & neck, colorectal, pancreatic, lung [4], neuroblastomas [9], and CD20+ NHL [7]. ATC armed with HER2Bi were not only able to lyse cancer cells that have high (3+) expression of HER2 but more importantly target and lyse MCF-7 cells that express low or nil HER2 expression [6] Moreover, armed ATC can kill multiple times, secrete cytokines/chemokines and multiply after engaging tumor cells in vitro[10]. "
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