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Figure 1 | Retrovirology

Figure 1

From: Interferon-α (IFN-α) suppresses HTLV-1 gene expression and cell cycling, while IFN-α combined with zidovudin induces p53 signaling and apoptosis in HTLV-1-infected cells

Figure 1

Effects of IFN-α treatment on HTLV-1 p19 release and viral transcription in various HTLV-1-infected cell lines. A. Expression of HTLV-1 mRNAs (a) and proteins (b, c) were evaluated by quantitative RT-PCR (a), immunoblotting (b), and flow cytometry (c), respectively, in HTLV-1-infected HUT102, ILT-Hod and ILT-#29 or uninfected Jurkat cell lines. a. The mRNA copy numbers measured by using pX or Gag primers were standardized to those for GAPDH and indicated as the means and standard deviations (SD) of duplicate samples. b. Cell lysates from indicated cell lines were subjected to an immunoblotting assay with antibodies to Tax (40 kDa) and α-Tubulin (50 kDa). The lysates in lanes 5 and 6 were prepared from ILT-Hod and ILT-#29 cells stimulated with PMA (50 ng/ml) overnight, respectively. c. Intracellular Tax proteins in permeabilized cells were stained with Alexa Fluor 488-labeled anti-Tax mAb (open histogram) and mouse IgG3 isotype control antibody (closed histogram). The inserted box indicates Gag expression in ILT-Hod and ILT-#29 cells stimulated with PMA (50 ng/ml) for 17h. B. HUT102 (top), ILT-Hod (middle) and ILT-#29 (bottom) cells were cultured for 3 days with or without three doses of IFN-α indicated. HTLV-1 p19 concentrations in the supernatants (left) and Gag mRNA levels were measured by ELISA and quantitative RT-PCR, respectively. Data are presented as the means and SD of duplicate samples. C. Frozen stored primary ATL cells were thawed and analyzed for intracellular Tax (top) or Gag (bottom) proteins by flow cytometry immediately (green line) or 24 h after culture with no (black line), 300 IU/ml (red line) or 3000 IU/ml (blue line) of IFN-α in the presence of IL-2 (30 IU/ml). The closed histogram represents samples stained with isotype controls. The mean fluorescence intensity (MFI) of each histogram was indicated in the bar graphs.

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