A new sensitive and quantitative HTLV-I-mediated cell fusion assay in T cells.
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-kappaB 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.
Paré
Marie-Eve
ME
Centre de Recherche en Infectiologie, Centre Hospitalier Universitaire de Québec, Pavillon CHUL, Canada.
Gauthier
Sonia
S
Landry
Sébastien
S
Sun
Jiangfeng
J
Legault
Eric
E
Leclerc
Denis
D
Tanaka
Yuetsu
Y
Marriott
Susan J
SJ
Tremblay
Michel J
MJ
Barbeau
Benoit
B
D015496:CD4-Positive T-Lymphocytes / Q000276:immunology* / Q000821:virology*; D002459:Cell Fusion; D002460:Cell Line; D018920:Coculture Techniques; D015368:Human T-lymphotropic virus 1 / Q000235:genetics / Q000276:immunology*; D006801:Humans; D019169:Jurkat Cells; D015183:Restriction Mapping; D012680:Sensitivity and Specificity; D014771:Virion / Q000276:immunology