Flow cytometric methodology for the detection of de novo human T-cell leukemia virus -1 infection in vitro: A tool to study novel infection inhibitors

Highlights

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  MT-2 cells express more Tax, gp46 and p19 than HUT102′s.

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  HUT78 cells express higher levels of the HTLV-1 permissive receptors neuropilin and GLUT-1 than CEM or JURKAT.

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  Irradiation does not eliminate all MT-2 donor cells in HTLV-1 co-culture protocols.

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  Flow cytometry and Lt-4 anti-tax antibody can detect de novo HTLV-1 infection at early time points.

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  Cytochalasin B and sodium valproate inhibit HTLV-1 infection at early time points.

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.