A Suppressive Effect of Cyclosporine A on Replication and Noncoding Control Region Activation of Polyomavirus BK Virus

Abstract

The effect of cyclosporine A (CsA) on polyomavirus BK virus (BKV) replication remains unclear. The aim of this in vitro study was to investigate the effect of CsA on BKV replication in human uroepithelial cells. After infection of a human renal proximal tubular cell line, HK-2 with BKV, BKV viral load in the presence of CsA was assessed by real-time polymerase chain reaction. The BKV large T-antigen (LTag) expression was measured by Western blot analysis. The BKV early promoter activity was determined by measuring luciferase activity of the BKV noncoding control region luciferase reporter. The BKV LTag expression in a human bladder carcinoma cell line, T24, was assessed by immunofluorescence. The results demonstrated that the increased levels of BKV LTag and viral protein 1 transcripts measured by real-time polymerase chain reaction were suppressed by CsA in a dose-dependent manner (0.5-4 microg/mL). Western blot analysis also showed that CsA inhibited BKV LTag expression. In addition, the activity of the BKV early promoter, which was enhanced by BKV LTag overexpression, was abrogated by CsA. Finally, the suppressive effect of CsA on BKV replication was also shown in T24 cells as CsA reduced immunofluorescent staining of BKV LTag in these cells. This in vitro study indicates that CsA suppresses BKV replication in human proximal renal tubular cells and uroepithelial cells of the urinary bladder and inhibits the BKV-LTag-regulated increase in early promoter activity.