Jeffers Liesl K, Madden Vicki, Webster-Cyriaque Jennifer
Department of Microbiology and Immunology, University of North Carolina, Chapel Hill, NC 27599, USA.
Virology. 2009 Nov 25;394(2):183-93. doi: 10.1016/j.virol.2009.07.022. Epub 2009 Sep 25.
In this study, it was determined that BKV is shed in saliva and an in vitro model system was developed whereby BKV can productively infect both submandibular (HSG) and parotid (HSY) salivary gland cell lines.
BKV was detected in oral fluids using quantitative real-time PCR (QRTPCR). BKV infection was determined using quantitative RT-PCR, immunofluorescence and immunoblotting assays. The infectivity of BKV was inhibited by pre-incubation of the virus with gangliosides that saturated the major capsid protein, VP1, halting receptor mediated BKV entry into salivary gland cells. Examination of infected cultures by transmission electron microscopy revealed 45-50 nm BK virions clearly visible within the cells. Subsequent to infection, encapsidated BK virus was detected in the supernatant.
We thus demonstrated that BKV was detected in oral fluids and that BK infection and replication occur in vitro in salivary gland cells. These data collectively suggest the potential for BKV oral route of transmission and oral pathogenesis.
在本研究中,已确定BK病毒(BKV)可在唾液中排出,并建立了一种体外模型系统,借此BKV可有效感染下颌下腺(HSG)和腮腺(HSY)唾液腺细胞系。
使用定量实时聚合酶链反应(QRT-PCR)在口腔液中检测到BKV。通过定量逆转录聚合酶链反应、免疫荧光和免疫印迹分析确定BKV感染情况。BKV与神经节苷脂预孵育可抑制其感染性,神经节苷脂可饱和主要衣壳蛋白VP1,阻止受体介导的BKV进入唾液腺细胞。通过透射电子显微镜检查受感染培养物,发现细胞内清晰可见45 - 50纳米的BK病毒粒子。感染后,在上清液中检测到包封的BK病毒。
因此,我们证明在口腔液中检测到BKV,且BK病毒在唾液腺细胞中可进行体外感染和复制。这些数据共同表明BKV经口腔传播和口腔发病机制的可能性。