A new sensitive and quantitative HTLV-I-mediated cell fusion assay in T cells
| Autor: | Jiangfeng Sun, Éric Legault, Michel J. Tremblay, Benoit Barbeau, Yuetsu Tanaka, Denis Leclerc, Susan J. Marriott, Sonia Gauthier, Marie-Ève Paré, Sébastien Landry |
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| Rok vydání: | 2005 |
| Předmět: |
CD4-Positive T-Lymphocytes
T cell viruses Cell Tax Restriction Mapping Biology Jurkat cells Sensitivity and Specificity Cell Line Cell Fusion Jurkat Cells immune system diseases Virology medicine Humans Syncytium Human T-lymphotropic virus 1 Cell fusion Virion virus diseases Transfection HTLV-I Molecular biology Coculture Techniques medicine.anatomical_structure Cell culture Giant cell Luciferase |
| Zdroj: | Virology. 338(2):309-322 |
| ISSN: | 0042-6822 |
| DOI: | 10.1016/j.virol.2005.05.012 |
| Popis: | Similar to several other viruses, human T cell leukemia virus type I (HTLV-I) induces the formation of multinucleated giant cells (also known as syncytium) when amplified in tissue culture. These syncytia result from the fusion of infected cells with uninfected cells. Due to the intrinsic difficulty of infecting cells with cell-free HTLV-I virions, syncytium formation has become an important tool in the study of HTLV-I infection and transmission. Since most HTLV-I-based cell fusion assays rely on the use of non-T cells, the aim of this study was to optimize a new HTLV-I-induced cell fusion assay in which HTLV-I-infected T cell lines are co-cultured with T cells that have been transfected with an HTLV-I long terminal repeat (LTR) luciferase reporter construct. We demonstrate that co-culture of various HTLV-I-infected T cells with different transfected T cell lines resulted in induction of luciferase activity. Cell-to-cell contact and expression of the viral gp46 envelope protein was crucial for this induction while other cell surface proteins (including HSC70) did not have a significant effect. This quantitative assay was shown to be very sensitive. In this assay, the cell fusion-mediated activation of NF-κB and the HTLV-I LTR occurred through previously described Tax-dependent signaling pathways. This assay also showed that cell fusion could activate Tax-inducible cellular promoters. These results thus demonstrate that this new quantitative HTLV-I-dependent cell fusion assay is versatile, highly sensitive, and can provide an important tool to investigate cellular promoter activation and intrinsic signaling cascades that modulate cellular gene expression. |
| Databáze: | OpenAIRE |
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