Neuroblastoma cells transiently transfected to simultaneously express the co-stimulatory molecules CD54, CD80, CD86, and CD137L generate antitumor immunity in mice. J Immunother 2005;28(5):449-60
Date
08/23/2005Pubmed ID
16113601DOI
10.1097/01.cji.0000171313.93299.74Scopus ID
2-s2.0-24144487135 (requires institutional sign-in at Scopus site) 32 CitationsAbstract
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.
Author List
Johnson BD, Gershan JA, Natalia N, Zujewski H, Weber JJ, Yan X, Orentas RJAuthor
Bryon D. Johnson PhD Adjunct Professor in the Medicine department at Medical College of WisconsinMESH terms used to index this publication - Major topics in bold
AnimalsAntigens, CD
Antineoplastic Agents
B7-1 Antigen
Cancer Vaccines
Cell Line, Tumor
Cell Proliferation
Electroporation
Flow Cytometry
Genetic Vectors
Humans
In Vitro Techniques
Intercellular Adhesion Molecule-1
Interferon-gamma
Mice
Mice, Inbred C57BL
Microscopy, Confocal
Neuroblastoma
Plasmids
Receptors, Nerve Growth Factor
Receptors, Tumor Necrosis Factor
Reverse Transcriptase Polymerase Chain Reaction
Spleen
T-Lymphocytes
Time Factors
Transcription, Genetic
Transfection
Tumor Necrosis Factor Receptor Superfamily, Member 9
