Redirected T-cell cytotoxicity to epithelial cell adhesion molecule-overexpressing adenocarcinomas by a novel recombinant antibody, E3Bi, in vitro and in an animal model.
BACKGROUND
To redirect cytotoxic T cells to target a broad range of adenocarcinomas, the authors constructed a novel, recombinant, bispecific antibody, E3Bi, directed at the tumor-associated antigen, epithelial cell adhesion molecule (EpCAM), and the CD3 receptor on T cells.


METHODS
T cells were prepared from healthy blood donors. The cytotoxicity of activated T cells (ATC) redirected to tumor cells by E3Bi was measured with in vitro (51)Cr release assays. In vivo studies were performed in a severe combined immunodeficient (SCID)/Beige mouse xenograft model. Tumor-bearing mice were treated with low doses (1 mg/kg) or high doses (10 mg/kg) of E3Bi along with ATC (2 x 10(9) cells/kg), and treatment efficacy was evaluated both by ex vivo tumor cell survival assay after in vivo treatments and by in vivo tumor growth delay studies.


RESULTS
In vitro, targeting the EpCAM-overexpressing human tumor cell lines with E3Bi increased specific cytotoxicity of ATC by > 70% at an effector-to-target ratio of 2.5 (P < 0.001); this cytotoxicity was abolished competitively in the presence of an anti-EpCAM monoclonal antibody. In contrast, E3Bi did not enhance ATC cytotoxicity toward the low EpCAM-expressing tumor cell line. In ex vivo tumor cytotoxicity assays, a significant reduction in tumor cell survival (40% with low-dose E3Bi; 90% with high-dose E3Bi) was observed in E3Bi/ATC-treated mice compared with control mice that were treated with ATC only. In addition, SCID/Beige mice xenografted with LS174T tumors demonstrated a significant tumor growth delay (P = 0.0139) after receiving E3Bi/ATC/interleukin 2 (IL-2) compared with mice that received ATC/IL-2 alone.


CONCLUSIONS
E3Bi specifically and very efficiently redirected T cells to destroy EpCAM-overexpressing tumors both in vitro and in an animal model. These results suggest a therapeutic utility for E3Bi in the treatment of adenocarcinomas.
Ren-Heidenreich
Lifen
L
Molecular Immunology Laboratory, Adele R. Decof Cancer Center, Roger Williams Hospital, Providence, Rhode Island 02908, USA. lifenren@yahoo.com
Davol
Pamela A
PA
Kouttab
Nicola M
NM
Elfenbein
Gerald J
GJ
Lum
Lawrence G
LG
D000230:Adenocarcinoma / Q000188:drug therapy / Q000276:immunology* / Q000473:pathology; D000818:Animals; D018033:Antibodies, Bispecific / Q000494:pharmacology*; D017209:Apoptosis; D015815:Cell Adhesion Molecules; D002470:Cell Survival; D003602:Cytotoxicity, Immunologic / Q000502:physiology*; D004195:Disease Models, Animal; D004847:Epithelial Cells / Q000187:drug effects / Q000276:immunology; D005260:Female; D066298:In Vitro Techniques; D008213:Lymphocyte Activation; D051379:Mice; D016513:Mice, SCID; D009374:Neoplasms, Experimental; D017260:Receptor-CD3 Complex, Antigen, T-Cell / Q000032:analysis / Q000276:immunology*; D012680:Sensitivity and Specificity; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology* / Q000502:physiology; D014407:Tumor Cells, Cultured
D018033:Antibodies, Bispecific; D015815:Cell Adhesion Molecules; D017260:Receptor-CD3 Complex, Antigen, T-Cell