Abstract
Methodology to detect and study de novo human T-cell leukemia virus (HTLV)-1 infection is required to further our knowledge of the viruses’ mechanisms of infection and to study potential therapeutic interventions. Whilst methodology currently exists, utilisation of an anti-Tax antibody to detect de novo Tax expression in permissive cells labelled with cell tracker allowing for the detection by flow cytometry of new infection after co-culture with donor cell lines productively infected with HTLV-1 is an alternative strategy. Using this methodology, we have been able to detect de novo infection of the T cell line HUT78 following co-culture with the productively infected HTLV-1 donor cell line MT-2 and to confirm that infection can be effectively blocked with well characterised infection inhibitors. This methodology will benefit experimental studies examining HTLV infection in vitro and may aid identification of therapeutic agents that block this process.
Highlights
Worldwide an estimated 10–15 million people live with human T-cell leukemia virus (HTLV)-1 (Gessain and Cassar, 2012), with areas of high endemicity in Japan, West Africa, South America, Caribbean islands, Romania, Iran, Middle East and Australo-Melanesia, where the prevalence ranges from 3 to 50% (Einsiedel et al, 2016; Gessain and Cassar, 2012; Murphy, 2016)
In vitro HTLV-1 infection studies and syncytium-assays are used to further our understanding of the mechanisms of HTLV-1 infection and to investigate novel drug inhibitors
A second productively infected HTLV-1 cell line, HUT102, was compared and, a small percentage survived to 240 h after 30 Gy irradiation
Summary
Worldwide an estimated 10–15 million people live with HTLV-1 (Gessain and Cassar, 2012), with areas of high endemicity in Japan, West Africa, South America, Caribbean islands, Romania, Iran, Middle East and Australo-Melanesia, where the prevalence ranges from 3 to 50% (Einsiedel et al, 2016; Gessain and Cassar, 2012; Murphy, 2016). There is currently no cure for HTLV-1 infection or its associated diseases and it has been considered a neglected virus. HTLV-1 is a milkand blood-borne and sexually transmitted virus causing significant mortality and morbidity in patients who develop disease such as HTLV associated myelopathy/Tropical Spastic Paraparesis (HAM/TSP) and adult T-cell leukemia/lymphoma (ATL), making HTLV-1 a significant threat to public health. In vitro HTLV-1 infection studies and syncytium-assays are used to further our understanding of the mechanisms of HTLV-1 infection and to investigate novel drug inhibitors. In vitro cell-free HTLV-1 infection has been reported (Fan et al, 1992; Jones et al, 2008), but is an inefficient process. Co-cultures of productively infected HTLV-1 donor cells with permissive cells are commonly used to study in vitro de novo infection (Feuer and Green, 2005). New infection is detected through polymerase chain reaction of HTLV-1 specific Tax
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