Vaccinia virus serves as an efficient vector for expressing heterologous proteins in human NTera 2 neurons.
The human teratocarcinoma cell line NTera 2 (NT2) can be induced to differentiate into post-mitotic neurons possessing well-defined axonal and dendritic morphology. Highly enriched neurons (NT2-N cells) can be prepared in large numbers, thus combining many of the advantages of both primary and continuous cell culture systems. Unfortunately, it has proven difficult to express foreign genes in NT2-N cells. We examined whether vaccinia virus (VV) can express heterologous proteins in NT2-N cells and characterized the response of NT2-N cells to VV infection. NT2-N cells were infected with VV vectors expressing the envelope glycoprotein (gp160) from the human immunodeficiency type 1 virus (HIV 1). These vectors were chosen because VV-directed synthesis and post-translational processing of gp160 have been well characterized in many cell types. Approximately 85% of the neurons expressed gp160 which underwent native post-translational cleavage. The rate of gp160 synthesis was maximal at 5-48 hours postinfection, but was detectable for as long as 4 days. Surprisingly, NT2-N cells showed no VV-induced alterations in morphology, downregulation of host protein synthesis, or cytotoxicity, as measured by lactate dehydrogenase release. These results indicate that VV can serve as an efficient vector for introducing foreign genes in NT2-N cells without the cytotoxic effects often associated with VV infection. These properties, in conjunction with the advantages provided by NT2-N cells, provide new options for analyzing the cellular and molecular functions of human neurons.
Department of Pathology, University of Pennsylvania, Philadelphia 19104, USA.
D G
DG
Cook
R S
RS
Turner
D L
DL
Kolson
V M
VM
Lee
R W
RW
Doms
D016923:Cell Death / Q000502:physiology; D002478:Cells, Cultured; D004279:DNA, Viral / Q000096:biosynthesis; D005822:Genetic Vectors; D019302:HIV Envelope Protein gp160 / Q000096:biosynthesis; D006801:Humans; D008854:Microscopy, Electron; D009419:Nerve Tissue Proteins / Q000096:biosynthesis* / Q000235:genetics*; D009474:Neurons / Q000378:metabolism* / Q000648:ultrastructure; D013936:Thymidine / Q000378:metabolism; D014407:Tumor Cells, Cultured; D014616:Vaccinia virus
D004279:DNA, Viral; D019302:HIV Envelope Protein gp160; D009419:Nerve Tissue Proteins; D013936:Thymidine