Establishment of a biological assay system for human retroviral protease activity.
In order to obtain indicator cell lines that are exquisitely susceptible to human T-lymphotropic virus type 1 (HTLV-1), luciferase gene driven by HTLV-1 long terminal repeat (LTR) was transfected into lymphocytic H9 cells with neo gene, and cell lines were selected by G418. A cell line (H9/K30luc) was found to produce an extremely high level of luciferase only when co-cultured with HTLV-1 producer MT-2 cells. Both in the absence and presence of a reverse transcriptase (RT) inhibitor azidothymidine, H9/K30luc cells generated similarly high luciferase activity upon co-cultivation with MT-2 cells. To develop an equivalent system for human immunodeficiency virus type 1 (HIV-1), H9/NL432 cells, which are stably infected with HIV-1 and producing a low level of the virus-like MT-2 cells for HTLV-1, were generated. Together with the indicator cell line H9/H1luc for HIV-1 already reported, antiviral effects of some agents on HTLV-1 and HIV-1 could be readily and sensitively evaluated by similar methods. In fact, by using our system, an HIV-1 protease inhibitor, saquinavir, was demonstrated to be highly effective against HIV-1 but not against HTLV-1.
Yoshida
Akiko
A
Department of Virology, Institute of Health Biosciences, The University of Tokushima Graduate School, 3-18-15 Kuramoto-cho, Tokushima-shi, Tokushima 770-8503, Japan. akko@basic.med.tokushima-u.ac.jp
Piroozmand
Ahmad
A
Sakurai
Akiko
A
Fujita
Mikako
M
Uchiyama
Tsuneo
T
Kimura
Tooru
T
Hayashi
Yoshio
Y
Kiso
Yoshiaki
Y
Adachi
Akio
A
D000998:Antiviral Agents / Q000494:pharmacology; D002460:Cell Line; D018920:Coculture Techniques / Q000379:methods; D015870:Gene Expression; D017930:Genes, Reporter; D015497:HIV-1 / Q000187:drug effects / Q000502:physiology; D015368:Human T-lymphotropic virus 1 / Q000187:drug effects / Q000502:physiology*; D006801:Humans; D008156:Luciferases / Q000235:genetics* / Q000378:metabolism*; D008214:Lymphocytes / Q000821:virology*; D019258:Saquinavir / Q000494:pharmacology
D000998:Antiviral Agents; D008156:Luciferases; D019258:Saquinavir