A novel medium-throughput biological assay system for HTLV-۱ infectivity and drug discovery

Objective(s): Here, a reporter cell line containing two reporter vectors were developed, in order to monitor the Human T-Lymphotropic Virus type۱(HTLV-۱) infectivity and the cell viability simultaneously. Materials and Methods: The reporter cell line was constructed by stably transfected baby hamster's kidney cell line (BHK-۲۱), with the genomes expressing two different reporters in separate plasmids.The first reporter gene is transactivated by the HTLV-۱ tax protein, while the second reporter is continuously expressed when introduced into a mammalian cell. In order to show its functionality, the effect of the drug mix on HTLV-۱ was assayed by this system and was compared to the results obtained by other methods. Results: HTLV-۱ reporter cell line was found to produce high level of luciferase when co-cultured with MT-۲ and Hut-۱۰۲ cells but not with Jurkat cell. Moreover, the combination therapy against HTLV-۱ can reduce luciferase expression of the cell when co-cultured with MT-۲ and Hut-۱۰۲ comparable to the ELISA (R=۰.۹۳۲, P-value =۰.۰۰۲). In addition, the results revealed the superiority of the present system over the molecular methods. Conclusion: The results demonstrated that the biological assay system is a beneficial tool for the medium-throughput anti-HTLV-۱ drug screening and inhibitory effect.

Keywords:

Biological Assay , Drug screening , HTLV-۱ , Hut۱۰۲ , Luciferase , Reporter gene