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J. Virol. doi:10.1128/JVI.02538-06
Copyright (c) 2007, American Society for Microbiology and/or the Listed Authors/Institutions. All Rights Reserved.

Construction and characterization of a full length infectious Simian T-cell Lymphotropic Virus Type-3 molecular clone

Unité d'Epidémiologie et Physiopathologie des Virus Oncogènes, CNRS URA 3015, Département de Virologie, Institut Pasteur, 28 rue du Dr Roux, 75015 Paris, France; Plateforme de Microscopie Electronique, Institut Pasteur, 25 rue du Dr Roux, 75015 Paris, France

^* To whom correspondence should be addressed. Email: rmahieux{at}pasteur.fr.

	Abstract

Together with their simian STLV equivalent, HTLV-1, HTLV-2 and HTLV-3 form the Primate T Lymphotropic virus (PTLV) group. Over years, understanding the HTLV-1 and HTLV-2 biology and their pathogenesis has been widely improved by the creation of molecular clones. On the contrary, so far, PTLV-3 experimental studies have been restricted to overexpression of tax gene using reporter assays. We have therefore decided to construct an STLV-3 molecular clone. We generated a full length STLV-3 proviral clone (8891 bp) by PCR amplification of overlapping fragments. This STLV-3 molecular clone was then transfected into 293T cells. RT-PCR experiments followed by sequence analysis of the amplified products allowed us to establish that both gag and tax/rex mRNAs were transcribed. Western-blot further demonstrated the presence of the STLV-3 p24^gag protein in the cell culture supernatant from transfected cells. Transient transfection of 293T and of 293T-LTR-GFP cells with the STLV-3 clone promoted syncytia formation, a hallmark of PTLV Env expression, together with the appearance of fluorescent cells, also demonstrating that the Tax-3 protein is expressed. Virus particles were visible by electron microscopy. These particles are infectious, as demonstrated by our cell-free infection experiments with purified virions. Altogether, our data demonstrate that the STLV-3 molecular clone is functional and infectious. This clone will give us a unique opportunity to study in vitro the different pX transcripts, the putative presence of antisense transcripts and to evaluate the PTLV-3 pathogenicity in vivo.

This article has been cited by other articles:

• Chevalier, S. A., Ko, N. L., Calattini, S., Mallet, A., Prevost, M.-C., Kehn, K., Brady, J. N., Kashanchi, F., Gessain, A., Mahieux, R. (2008). Construction and Characterization of a Human T-Cell Lymphotropic Virus Type 3 Infectious Molecular Clone. J. Virol. 82: 6747-6752 [Abstract] [Full Text]