Neuroblastoma cells transiently transfected to simultaneously express the co-stimulatory molecules CD54, CD80, CD86, and CD137L generate antitumor immunity in mice.
The goal of this study was to show that nonviral gene transfection technology can be used to genetically modify neuroblastoma cells with immune stimulatory molecules, and that the modified cells can generate an antitumor immune response. The authors found that an electroporation-based gene transfection method, nucleofection, could be used to modify mouse AGN2a (an aggressive variant of Neuro-2a) neuroblastoma cells to simultaneously express as many as four different immune stimulatory molecules encoded by separate plasmid vectors. Within 18 hours after nucleofection, greater than 60% of the cells typically expressed the transfected gene products, and the percentages of cells expressing the products often exceeded 96%. High levels of plasmid in cell nuclei immediately after nucleofection documented instantaneous availability of gene vectors to the transcriptional machinery. AGN2a cells nucleofected to express the co-stimulatory molecules CD80 and CD86 expressed higher levels of these molecules than cells that had been permanently transfected with these same plasmid vectors, and the nucleofected cells were as effective as the permanently transfected cells at inducing an antitumor response in vivo in a tumor prevention model. AGN2a cells nucleofected with four separate plasmid vectors encoding CD54, CD80, CD86, and CD137L induced a T-cell immune response in vitro and served as a potent tumor vaccine in the tumor prevention model. These data show that transient transfection using a nonviral based method, nucleofection, can be used to rapidly generate novel cell-based tumor vaccines.
Johnson
Bryon D
BD
Department of Pediatrics, Medical College of Wisconsin and Children's Research Institute, Children's Hospital of Wisconsin, Milwaukee, WI 53226, USA. bjohnson@mcw.edu
Gershan
Jill A
JA
Natalia
Natalia
N
Zujewski
Heidi
H
Weber
James J
JJ
Yan
Xiaocai
X
Orentas
Rimas J
RJ
D000818:Animals; D015703:Antigens, CD / Q000096:biosynthesis*; D000970:Antineoplastic Agents / Q000494:pharmacology; D018122:B7-1 Antigen / Q000096:biosynthesis*; D019496:Cancer Vaccines / Q000378:metabolism; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D018274:Electroporation; D005434:Flow Cytometry; D005822:Genetic Vectors; D006801:Humans; D066298:In Vitro Techniques; D018799:Intercellular Adhesion Molecule-1 / Q000096:biosynthesis*; D007371:Interferon-gamma / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D018613:Microscopy, Confocal; D009447:Neuroblastoma / Q000378:metabolism*; D010957:Plasmids / Q000378:metabolism; D017475:Receptors, Nerve Growth Factor / Q000096:biosynthesis*; D018124:Receptors, Tumor Necrosis Factor / Q000096:biosynthesis*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013154:Spleen / Q000166:cytology; D013601:T-Lymphocytes / Q000378:metabolism; D013997:Time Factors; D014158:Transcription, Genetic; D014162:Transfection; D053261:Tumor Necrosis Factor Receptor Superfamily, Member 9
D015703:Antigens, CD; D000970:Antineoplastic Agents; D018122:B7-1 Antigen; D019496:Cancer Vaccines; D017475:Receptors, Nerve Growth Factor; D018124:Receptors, Tumor Necrosis Factor; C505714:TNFRSF9 protein, human; C505715:Tnfrsf9 protein, mouse; D053261:Tumor Necrosis Factor Receptor Superfamily, Member 9; D018799:Intercellular Adhesion Molecule-1; D007371:Interferon-gamma