title: Expression of HTLV-1 Genes in T-Cells Using RNA Electroporation.
abstract: Human T-cell leukemia virus type 1 (HTLV-1) infects about 20 million people world-wide. Around 5% of the infected individuals develop adult T-cell leukemia (ATL) or a neurological disease termed tropical spastic paraparesis (TSP) after a clinical latency of years to decades. Through the use of two promoters and alternative splicing HTLV-1 expresses at least 12 different proteins. HTLV-1 establishes a life-long persistent infection by inducing the clonal expansion of infected cells, a property largely ascribed to the viral genes Tax and HBZ. However, the fact that ATL arises in a minority of infected individuals after a long clinical latency suggests the existence of factors counterbalancing the oncogenic potential of HTLV-1 in the context of natural infection.To study the role of the different HTLV-1 gene products in the HTLV-1 life cycle, we optimized a transfection protocol for primary T-cells using an approach based on the electroporation of in vitro-transcribed RNA. Results showed that the RNA transfection technique combines a high transfection efficiency with low toxicity, not only in Jurkat T-cells but also in primary T-cells. These findings suggest that RNA electroporation is preferable for experiments aimed at investigating the role of HTLV-1 gene products in the context of primary T-cells, which represent the main target of HTLV-1 in vivo.
authors.affiliation: Department of Surgery, Oncology and Gastroenterology, University of Padova, Via Gattamelata 64, Padova, 35128, Italy.
authors.forename: Mariangela
authors.initials: M
authors.lastname: Manicone
authors.affiliation: Department of Surgery, Oncology and Gastroenterology, University of Padova, Via Gattamelata 64, Padova, 35128, Italy.
authors.forename: Francesca
authors.initials: F
authors.lastname: Rende
authors.affiliation: Istituto Oncologico Veneto, IRCCS, Via Gattamelata 64, Padova, 35128, Italy.
authors.forename: Ilaria
authors.initials: I
authors.lastname: Cavallari
authors.affiliation: Institute of Clinical and Molecular Virology, Friedrich-Alexander-Universität Erlangen-Nürnberg, Schlossgarten 5, Erlangen, 91054, Germany.
authors.forename: Andrea K
authors.initials: AK
authors.lastname: Thoma-Kress
authors.affiliation: Department of Surgery, Oncology and Gastroenterology, University of Padova, Via Gattamelata 64, Padova, 35128, Italy. v.ciminale@unipd.it.|Istituto Oncologico Veneto, IRCCS, Via Gattamelata 64, Padova, 35128, Italy. v.ciminale@unipd.it.
authors.forename: Vincenzo
authors.initials: V
authors.lastname: Ciminale
mesh_terms: D050976:Basic-Leucine Zipper Transcription Factors / Q000096:biosynthesis / Q000235:genetics; D018274:Electroporation; D015870:Gene Expression; D016356:Gene Products, tax / Q000096:biosynthesis / Q000235:genetics; D005814:Genes, Viral; D015368:Human T-lymphotropic virus 1 / Q000235:genetics / Q000378:metabolism; D006801:Humans; D019169:Jurkat Cells; D012367:RNA, Viral / Q000235:genetics / Q000378:metabolism; D012191:Retroviridae Proteins / Q000096:biosynthesis / Q000235:genetics
chemical_list: D050976:Basic-Leucine Zipper Transcription Factors; D016356:Gene Products, tax; C479751:HBZ protein, human T-cell leukemia virus type I; D012367:RNA, Viral; D012191:Retroviridae Proteins; C514970:tax protein, Human T-lymphotrophic virus 1
keywords: ATL; HTLV-1; In vitro transcription; Peripheral blood mononuclear cells (PBMCs); RNA electroporation