Targeting immune effector molecules to human tumor cells through genetic delivery of 5T4-specific scFv fusion proteins.
Although several clinical trials have shown beneficial effects by targeting tumor-associated antigens (TAAs) with monoclonal antibodies, a number of issues, including poor penetration of the tumor mass and human antimouse antibody responses, remain. The use of recombinant single-chain Fv (scFv) fragments has the potential to address these and other issues while allowing the addition of different effector functions. To develop therapeutic strategies that recruit both humoral and cellular arms of the immune response, we have constructed chimeric proteins linking either the human IgG1 Fc domain or the extracellular domain of murine B7.1 to a scFv specific for the oncofetal glycoprotein, 5T4. This TAA is expressed by a wide variety of carcinomas and is associated with metastasis and poorer clinical outcome. We have engineered retroviral constructs that produce fusion proteins able to interact simultaneously with both 5T4-positive cells and with the receptor/ligands of the immune effector moieties. Genetic delivery through a murine leukemia virus vector to 5T4-positive tumor cells results in the secreted scFv fusion protein binding to the cell surface. Furthermore, the scFv-HIgG1 fusion protein is able to direct lysis of 5T4-expressing human tumor cell lines through antibody-dependent cell cytotoxicity, indicating its potential as a gene therapy for human cancers.
Myers
Kevin A
KA
Oxford BioMedica (UK) Ltd., Medawar Centre, Oxford Science Park, UK.
Ryan
Matthew G
MG
Stern
Peter L
PL
Shaw
David M
DM
Embleton
M Jim
MJ
Kingsman
Susan M
SM
Carroll
Miles W
MW
D000920:Antibody-Dependent Cell Cytotoxicity; D000951:Antigens, Neoplasm / Q000276:immunology*; D001483:Base Sequence; D002460:Cell Line; D005434:Flow Cytometry; D006801:Humans; D007135:Immunoglobulin Variable Region / Q000235:genetics / Q000276:immunology; D007145:Immunoglobulin kappa-Chains / Q000235:genetics / Q000276:immunology; D007167:Immunotherapy / Q000379:methods; D007668:Kidney; D008969:Molecular Sequence Data; D011993:Recombinant Fusion Proteins / Q000032:analysis; D014162:Transfection; D014407:Tumor Cells, Cultured
D000951:Antigens, Neoplasm; D007135:Immunoglobulin Variable Region; D007145:Immunoglobulin kappa-Chains; D011993:Recombinant Fusion Proteins