Glorieux Sarah, Favoreel H W, Meesen G, de Vos W, Van den Broeck W, Nauwynck H J
Laboratory of Virology, Faculty of Veterinary Medicine, Ghent University, Salisburylaan 133, B-9820 Merelbeke, Belgium.
Vet Microbiol. 2009 May 12;136(3-4):341-6. doi: 10.1016/j.vetmic.2008.11.005. Epub 2008 Nov 18.
Different alphaherpesviruses, including pseudorabies virus (PRV), are able to cross the basement membrane barrier in nasal respiratory epithelium. As a first step in investigating this invasion process, a detailed quantitative analysis system was set up to determine the kinetics of horizontal and vertical virus spread in nasal explants, using the virulent PRV strain 89V87. Plaque latitudes, total depths, depths measured from the basement membrane and volumes were determined at 0, 12, 24 and 36h post inoculation (pi). PRV 89V87 was found to spread in a plaquewise manner and to cross the basement membrane between 12 and 24hpi. During the 1960s-1970s, an increase in PRV virulence has been reported. To analyse potential differences in efficiency of infection and spread for different historical PRV strains, single infected cells and plaques of infected cells were quantified at 12 and 36hpi in nasal mucosa explants for seven European PRV strains, isolated in the 1960s (Becker, NIA1), the 1970s (NS374, NIA3, 75V19) and later (89V87, 00V72). All viruses were used at second passage in cell culture, except for the Becker strain, which had an unknown passage history. Older strains, Becker, NIA1 and/or NS374, showed lower numbers of primary infectious centers, lower capacity to form plaques and/or lower capacity to cross the basement membrane. The observed differences in virus-mucosa interactions may aid in understanding the virulence increase of PRV. The quantitative assay established here will be of use in unravelling the mechanism of alphaherpesvirus-mediated invasion through the basement membrane.
包括伪狂犬病病毒（PRV）在内的不同甲型疱疹病毒能够穿过鼻呼吸上皮中的基底膜屏障。作为研究这一侵袭过程的第一步，我们建立了一个详细的定量分析系统，以确定强毒PRV毒株89V87在鼻外植体中水平和垂直病毒传播的动力学。在接种后0、12、24和36小时（pi）测定噬斑纬度、总深度、从基底膜测量的深度和体积。发现PRV 89V87以噬斑方式传播，并在接种后12至24小时穿过基底膜。在20世纪60年代至70年代，有报道称PRV毒力有所增加。为了分析不同历史PRV毒株在感染和传播效率方面的潜在差异，在接种后12和36小时对七种欧洲PRV毒株（分别于20世纪60年代分离的Becker、NIA1，20世纪70年代分离的NS374、NIA3、75V19，以及后来分离的89V87、00V72）的鼻黏膜外植体中的单个感染细胞和感染细胞噬斑进行了定量。除Becker毒株传代历史不明外，所有病毒均在细胞培养中传代两次后使用。较老的毒株，如Becker、NIA1和/或NS374，显示出初级感染中心数量较少、形成噬斑的能力较低和/或穿过基底膜的能力较低。观察到的病毒与黏膜相互作用的差异可能有助于理解PRV毒力的增加。这里建立的定量测定方法将有助于阐明甲型疱疹病毒介导的穿过基底膜的侵袭机制。
