title: Quantitative Analysis of Human T-Lymphotropic Virus Type 1 (HTLV-1) Infection Using Co-Culture with Jurkat LTR-Luciferase or Jurkat LTR-GFP Reporter Cells.
abstract: Unlike HIV-1, HTLV-1 viral transmission requires cell-to-cell contacts, while cell-free virions are poorly infectious and almost absent from body fluids. Though the virus uses three nonexclusive mechanisms to infect new target cells: (1) MTOC polarization followed by formation of a virological synapse and viral transfer into a synaptic cleft, (2) genesis of a viral biofilm and its transfer of embedded viruses, or (3) HTLV-1 transmission using conduits. The Tax transactivator and the p8 viral proteins are involved in virological synapse and nanotube formation respectively.HTLV-1 transcription from the viral promoter (i.e., LTR) requires the Tax protein that is absent from the viral particle and is expressed after productive infection. The present chapter focuses on a series of protocols used to quantify HTLV-1 de novo infection of target cells. These techniques do not discriminate between the different modes of transmission, but allow an accurate measure of productive infection. We used cell lines that are stably transfected with LTR-GFP or LTR-luciferase plasmids and quantified Green Fluorescent Protein expression or luciferase activity, since both of them reflect Tax expression.
authors.affiliation: Equipe Oncogenèse Rétrovirale, Lyon, 69364 Cedex, 07, France.|Equipe labellisée "Ligue Nationale Contre le Cancer", Lyon, 69364 Cedex, 07, France.|International Center for Research in Infectiology, INSERM U1111 - CNRS UMR5308, Lyon, 69364 Cedex, 07, France.|Ecole Normale Supérieure de Lyon, 5 parvis René Descartes, BP 7000, 69342, Lyon, Cedex 07, France.|Université Lyon 1, Lyon, 69364 Cedex, 07, France.
authors.forename: Sandrine
authors.initials: S
authors.lastname: Alais
authors.affiliation: Equipe Oncogenèse Rétrovirale, Lyon, 69364 Cedex, 07, France.|Equipe labellisée "Ligue Nationale Contre le Cancer", Lyon, 69364 Cedex, 07, France.|International Center for Research in Infectiology, INSERM U1111 - CNRS UMR5308, Lyon, 69364 Cedex, 07, France.|Ecole Normale Supérieure de Lyon, 5 parvis René Descartes, BP 7000, 69342, Lyon, Cedex 07, France.|Université Lyon 1, Lyon, 69364 Cedex, 07, France.
authors.forename: Hélène
authors.initials: H
authors.lastname: Dutartre
authors.affiliation: Equipe Oncogenèse Rétrovirale, Lyon, 69364 Cedex, 07, France. renaud.mahieux@ens-lyon.fr.|Equipe labellisée "Ligue Nationale Contre le Cancer", Lyon, 69364 Cedex, 07, France. renaud.mahieux@ens-lyon.fr.|International Center for Research in Infectiology, INSERM U1111 - CNRS UMR5308, Lyon, 69364 Cedex, 07, France. renaud.mahieux@ens-lyon.fr.|Ecole Normale Supérieure de Lyon, 5 parvis René Descartes, BP 7000, 69342, Lyon, Cedex 07, France. renaud.mahieux@ens-lyon.fr.|Université Lyon 1, Lyon, 69364 Cedex, 07, France. renaud.mahieux@ens-lyon.fr.
authors.forename: Renaud
authors.initials: R
authors.lastname: Mahieux
mesh_terms: D018920:Coculture Techniques; D016356:Gene Products, tax / Q000235:genetics / Q000378:metabolism*; D017930:Genes, Reporter; D015490:HTLV-I Infections / Q000235:genetics / Q000378:metabolism* / Q000635:transmission*; D015368:Human T-lymphotropic virus 1 / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D019169:Jurkat Cells; D008156:Luciferases / Q000096:biosynthesis* / Q000235:genetics
chemical_list: D016356:Gene Products, tax; C514970:tax protein, Human T-lymphotrophic virus 1; D008156:Luciferases
keywords: Biofilm; Cell-to-cell infection; Flow cytometry; GFP; HTLV-1; Luciferase assay; Tax; Virological synapse