In vitro targeting of NG2 antigen by 213Bi-9.2.27 alpha-immunoconjugate induces cytotoxicity in human uveal melanoma cells.
OBJECTIVE
To examine uveal melanoma cell lines for the expression of human melanoma proteoglycan (NG2) using monoclonal antibody (mAb) 9.2.27 and subsequently to assess the in vitro specificity and cytotoxicity of mAb 9.2.27 conjugated to the alpha-particle-emitting radioisotope 213bismuth (213Bi-9.2.27) for uveal melanoma cells.


METHODS
Immunocytochemistry and flow cytometry were used to examine OCM-1, OCM-3, OCM-8, OMM-1, Mel202 and 92-1 melanoma cell lines for NG2 expression. Melanoma cells were treated with test (213Bi-9.2.27) or control (213Bi-A2) alpha-immunoconjugates (AICs). The specific cytotoxicity of 213Bi-9.2.27 AIC was evaluated using an MTS (3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfenyl)-2H-tetrazolium, inner salt) assay. Cell death was also assessed using TUNEL.


RESULTS
OCM-1, OCM-8, OMM-1, and Mel202 cells strongly expressed NG2. OCM-3 cells showed moderate expression and 92-1 cells were NG2-negative. 213Bi-9.2.27 specifically killed NG2-positive OCM-1, OCM-8, and OMM-1 cells in a concentration-dependent manner. D0 values for 37% cell survival of NG2-positive OCM-1, OCM-8, and OMM-1 cells were 5.8, 5.0, and 5.6 microCi, respectively, and the value was 43.4 muCi for NG2-negative 92-1 cells.


CONCLUSIONS
The specific cytotoxicity of 213Bi-9.2.27 AIC for NG2-positive, but not NG2-negative, cells suggests NG2 is a suitable target for alpha-immunotherapy in uveal melanoma. 213Bi-9.2.27 AIC used directly or as adjunct therapy may be a promising new agent for treating NG2-positive uveal melanomas or metastases.
Li
Yong
Y
Centre for Experimental Radiation Oncology, Cancer Care Centre, St. George Hospital, Kogarah, NSW, Australia.
Wang
Jian
J
Rizvi
Syed M Abbas
SM
Jager
Martine J
MJ
Conway
Robert M
RM
Billson
Francis A
FA
Allen
Barry J
BJ
Madigan
Michele C
MC
D018033:Antibodies, Bispecific / Q000627:therapeutic use*; D000941:Antigens / Q000276:immunology*; D001729:Bismuth / Q000627:therapeutic use*; D016923:Cell Death; D003602:Cytotoxicity, Immunologic; D004305:Dose-Response Relationship, Drug; D005434:Flow Cytometry; D006801:Humans; D018796:Immunoconjugates / Q000627:therapeutic use; D007150:Immunohistochemistry; D020287:In Situ Nick-End Labeling; D008545:Melanoma / Q000276:immunology / Q000473:pathology / Q000628:therapy*; D011509:Proteoglycans / Q000276:immunology*; D016499:Radioimmunotherapy; D011868:Radioisotopes / Q000627:therapeutic use*; D014407:Tumor Cells, Cultured; D014604:Uveal Neoplasms / Q000276:immunology / Q000473:pathology / Q000628:therapy*; D000099216:Chondroitin Sulfate Proteoglycan 4
D018033:Antibodies, Bispecific; D000941:Antigens; D001729:Bismuth; D018796:Immunoconjugates; D011509:Proteoglycans; D011868:Radioisotopes; D000099216:Chondroitin Sulfate Proteoglycan 4