Ex vivo delivery of suicide genes into melanoma cells using epidermal growth factor receptor-specific Fab immunogene.
The Fab fragment of monoclonal antibody B4G7 against human epidermal growth factor (EGF) receptor was conjugated with cationic poly-L-lysine and the resulting conjugate was further complexed with reporter genes or therapeutic genes. This Fab/DNA complex was designated as "Fab immunogene." The Fab immunogene transfer in vitro was mediated through the EGF receptors in two melanoma cell lines. The frequency of cells expressing beta-galactosidase (beta-Gal) reporter gene was approximately 1%. The induction of suicide effects after Fab immunogene transfer of herpes simplex virus thymidine kinase (TK) or Escherichia coli cytosine deaminase (CD) gene was quite remarkable, and the growth of melanoma cells was inhibited for over 7 days in the presence of ganciclovir (GCV) or 5-fluorocytosine (5-FC). Similarly, when melanoma cells treated in vitro with the Fab immunogene carrying TK or CD were transplanted into the back of nude mouse, subsequent systemic administration of GCV or 5-FC effectively suppressed the growth of tumors, indicating the occurrence of in vivo suicide effects.
Ohtake
Y
Y
Department of Molecular Biology, Keio University School of Medicine, Tokyo.
Chen
J
J
Gamou
S
S
Takayanagi
A
A
Mashima
Y
Y
Oguchi
Y
Y
Shimizu
N
N
D000818:Animals; D000918:Antibody Specificity; D066246:ErbB Receptors / Q000276:immunology*; D018014:Gene Transfer Techniques; D017930:Genes, Reporter; D015316:Genetic Therapy; D006801:Humans; D007125:Immunogenetics; D007140:Immunoglobulin Fab Fragments / Q000235:genetics*; D008545:Melanoma / Q000628:therapy*; D051379:Mice; D008819:Mice, Nude; D009368:Neoplasm Transplantation; D014407:Tumor Cells, Cultured; D001616:beta-Galactosidase / Q000235:genetics
D007140:Immunoglobulin Fab Fragments; D066246:ErbB Receptors; D001616:beta-Galactosidase