Selective gene transfer in vitro to tumor cells via recombinant Newcastle disease virus.
We developed a novel strategy to target recombinant Newcastle disease virus (NDV) to tumor cells for gene therapy. Modifying the virus with a bispecific fusion protein allowed virus receptor-independent tumor cell binding and gene transfer. The targeting molecule (alpha)HN-IL-2 contains an scFv antibody cloned from a neutralizing hemagglutinin-neuraminidase (HN)-specific hybridoma linked to the human cytokine IL-2. A recombinant NDV expressing the enhanced green fluorescent protein (NDFL-EGFP) was applied to show the expression of foreign genes in virus-infected tumor cells. At 24 hours after infection with the modified virus (NDFL-EGFP/(alpha)HN-IL-2), FACS analysis and fluorescence microscopy revealed neutralization of natural infection in IL-2 receptor-negative Jurkat leukemia cells, but targeted expression of EGFP in IL-2 receptor-positive human leukemia-derived MT-2 cells. The targeted gene delivery of NDFL-EGFP/(alpha)HN-IL-2 in MT-2 cells was blocked by the target ligand human IL-2. Selective virus entry to IL-2 receptor bearing tumor cells was also observed in a mixture of Jurkat and MT-2 cell lines. These results demonstrate that a recombinant NDV carrying a foreign gene can be successfully targeted to a specific tumor through a bispecific protein, which thereby increases the selectivity of gene transfer.
Bian
Huijie
H
Division of Cellular Immunology, German Cancer Research Center, D010, Im Neuenheimer Feld 280, 69120 Heidelberg, Germany.
Fournier
Philippe
P
Moormann
Rob
R
Peeters
Ben
B
Schirrmacher
Volker
V
D000818:Animals; D016466:CHO Cells; D045744:Cell Line, Tumor; D006224:Cricetinae; D003412:Cricetulus; D005434:Flow Cytometry; D005452:Fluoresceins; D018014:Gene Transfer Techniques; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors / Q000235:genetics / Q000627:therapeutic use*; D049452:Green Fluorescent Proteins / Q000378:metabolism; D015707:HN Protein / Q000235:genetics*; D006801:Humans; D007376:Interleukin-2 / Q000235:genetics; D008856:Microscopy, Fluorescence; D009369:Neoplasms / Q000235:genetics / Q000628:therapy*; D009522:Newcastle disease virus / Q000235:genetics*; D013388:Succinimides
C087165:5-(6)-carboxyfluorescein diacetate succinimidyl ester; D005452:Fluoresceins; D015707:HN Protein; D007376:Interleukin-2; D013388:Succinimides; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins