Functional difference in the HTLV-1 subgroup specific viral transcriptional regulators Tax and HBZ

Among HTLV-1-infected individuals, the lifetime risk of developing HTLV-1-associated myelopathy/tropical spastic paraparesis (HAM/TSP) is associated with the HTLV-1 gene subgroup (i.e., subgroup-A or -B) in Japan. The purpose of this study is to understand the molecular mechanism responsible for this association. Jurkat Tet-On 3G cells were transfected with each indicated plasmids (i.e., pTRE3G-IRES-empty, pTRE3G-Tax-A, pTRE3G-Tax-B, pTRE3G-HBZ-A, and pTRE3G-HBZ-B) using an electroporator and then cultured at 37ºC in RPMI1640 without antibiotics. After 24 h transfection, Tax or HBZ protein expression were induced by adding Dox (final concentration: 3 ng/ml) to the culture medium. After 24 h induction, cells were collected, and total RNA was extracted with the RNeasy Mini Kit (QIAGEN) following the manufacture’s instructions. We synthesize cDNA and performed microarray analysis according to the manufacturer's instructions (Agilent Tecnologies). Data were analyzed using GeneSpring GX (Agilent Technologies).