Jeffers-Francis Liesl K, Burger-Calderon Raquel, Webster-Cyriaque Jennifer
Department of Dental Research, School of Dentistry, University of North Carolina at Chapel Hill, United States.
Department of Microbiology and Immunology, School of Medicine, University of North Carolina at Chapel Hill, United States.
Antiviral Res. 2015 Jun;118:46-55. doi: 10.1016/j.antiviral.2015.02.002. Epub 2015 Mar 16.
BK polyomavirus (BKPyV) is a known kidney tropic virus that has been detected at high levels in HIV-associated salivary gland disease (HIV-SGD), one of the most important AIDS associated oral lesions. BKPyV has been detected in HIV-SGD patient saliva and replicates in salivary gland cells in vitro. BKPyV antivirals are currently in wide use to guard against BKPyV mediated organ rejection in kidney transplant recipients. The goal of this study was to investigate the inhibitory effects of three such antiviral agents, Ciprofloxacin, Cidofovir, and Leflunomide in BKPyV infected salivary gland cells. Human salivary gland cells, and Vero cells, were infected with BKPyV, treated with antiviral drugs and assessed for BKPyV gene expression and viral replication for up to 5 days post infection. The kinetics of BKPyV replication were different in salivary gland cells compared to kidney cells. Ciprofloxacin and Cidofovir had minimal effect on metabolic activity and host cell DNA replication, however, cell toxicity was detected at the protein level with Leflunomide treatment. Ciprofloxacin decreased BKV T Ag and VP1 mRNA expression by at least 50% in both cell types, and decreased T Ag protein expression at days 3 and 4 post infection. A 2.5-4 log decrease in intracellular DNA replication and a 2-3 log decrease in progeny release were detected with Ciprofloxacin treatment. Cidofovir and Leflunomide also inhibited BKPyV gene expression and DNA replication. The three drugs diminished progeny release by 30-90% and 2- to 6-fold decreases in infectious virus were detected post drug treatment by fluorescence focus assay. Additionally, three clinical BKPyV isolates were assessed for their responses to these agents in vitro. Cidofovir and Leflunomide, but not Ciprofloxacin treatment resulted in statistically significant inhibition of BKPyV progeny release from salivary gland cells infected with HIVSGD BKPyV isolates. All three drugs decreased progeny release from cells infected with a transplant derived viral isolate. In conclusion, treatment of human salivary gland cells with each of the three drugs produced modest decreases in BKPyV genome replication. These data highlight the need for continued studies to discover more effective and less toxic drugs that inhibit BKPyV replication in salivary gland cells.
BK多瘤病毒（BKPyV）是一种已知的嗜肾病毒，在HIV相关涎腺疾病（HIV-SGD）中被检测到高水平存在，HIV-SGD是最重要的与艾滋病相关的口腔病变之一。BKPyV已在HIV-SGD患者唾液中被检测到，并在体外涎腺细胞中复制。BKPyV抗病毒药物目前被广泛用于预防肾移植受者中BKPyV介导的器官排斥反应。本研究的目的是调查三种此类抗病毒药物环丙沙星、西多福韦和来氟米特对BKPyV感染的涎腺细胞的抑制作用。将人涎腺细胞和Vero细胞用BKPyV感染，用抗病毒药物处理，并在感染后长达5天评估BKPyV基因表达和病毒复制情况。与肾细胞相比，BKPyV在涎腺细胞中的复制动力学有所不同。环丙沙星和西多福韦对代谢活性和宿主细胞DNA复制影响最小，然而，来氟米特处理在蛋白质水平检测到细胞毒性。环丙沙星在两种细胞类型中均使BKV T抗原和VP1 mRNA表达至少降低50%，并在感染后第3天和第4天降低T抗原蛋白表达。环丙沙星处理后检测到细胞内DNA复制下降2.5 - 4个对数，子代释放下降2 - 3个对数。西多福韦和来氟米特也抑制BKPyV基因表达和DNA复制。这三种药物使子代释放减少30 - 90%，药物处理后通过荧光聚焦试验检测到感染性病毒减少2至6倍。此外，对三株临床BKPyV分离株进行了体外对这些药物反应的评估。西多福韦和来氟米特，但不是环丙沙星处理，导致从感染HIVSGD BKPyV分离株的涎腺细胞中BKPyV子代释放受到统计学显著抑制。所有三种药物均减少了从感染移植来源病毒分离株的细胞中的子代释放。总之，用这三种药物中的每一种处理人涎腺细胞都会使BKPyV基因组复制适度减少。这些数据突出了继续进行研究以发现更有效且毒性更小的抑制BKPyV在涎腺细胞中复制的药物的必要性。
