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Vaccinia Reporter Viruses for Quantifying Viral Function at All Stages of Gene Expression
Published on: May 15, 2014
Diagnostic tool based on an HTLV-1-Tax expression system in eukaryotic cells using a poxvirus vector
Jaqueline Gontijo de Souza1, Flávio Guimarães da Fonseca, Olindo Assis Martins-Filho
1Departamento de Microbiologia, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais - UFMG, Belo Horizonte, Brazil.
Abstract:
Human T-lymphotropic virus 1 (HTLV-1) induces an immune-mediated inflammatory disease affecting the nervous system that eventually is accompanied by ocular, rheumatic and dermatologic manifestations (HTLV-1 associated myelopathy/tropical spastic paraparesis, or HAM/TSP). Proviral load and HTLV-1 protein expression, mainly of Tax, is correlated with disease progression and induction of host-virus equilibrium breakdown that, reportedly, involves the presence of Tax-specific cytotoxic T lymphocytes (CTL), T regulatory cells and anti-Tax antibodies. Based on knowledge of anti-Tax antibodies as markers of disease progression, the objectives of this study were both to design an infection/transfection system using the Vaccinia virus and a tax-encoding plasmid for the expression of Tax protein as well as to use this cell support to evaluate anti-Tax IgG by flow cytometry. The flow cytometry assay was standardized using pooled sera from each test group (negative, asymptomatic and HAM/TSP patients). The HAM/TSP group presented higher IgG anti-Tax reactivity (above 70%) than the asymptomatic group (nearly 40% reactivity). The data indicate that the infection/transfection system is useful for assessing Tax expression. This is a promising assay for use as a diagnostic tool to detect IgG anti-Tax and monitor HTLV-1 infected individuals.
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