King D D, Brady B, Dodd D, Wust C J, Brown A
Department of Microbiology, College of Liberal Arts, University of Tennessee, Knoxville.
Arch Virol. 1988;102(1-2):49-61. doi: 10.1007/BF01315562.
Infection of the human leukemia hematopoietic stem cell line, K-562, with Semliki Forest virus (SFV) can be characterized by three stages: (1) an early virus-proliferating stage lasting 1 to 4 days post-infection (pi) in which infectious virus is produced in high titers (10(3)pfu/cell) but in which there is minimal cytopathic effect. All cells appear viable by trypan blue dye exclusion, although they do not proliferate, and DNA and cell protein synthesis decrease to less than 3% of uninfected controls within 24 hours; (2) an intermediate stage extending from day 5 to about day 24-30 pi in which the amount of infectious virus declines to low levels. During this stage, viral protein synthesis decreases to undetectable levels, although viral gylcoproteins are readily demonstrated by immunofluorescence and by immunoblot; however, capsid protein appears to degrade within 21 days pi. Cell numbers remain constant but the viability of the non-proliferating cells determined by trypan blue exclusion could not be determined with confidence; (3) a final long-term stage in which viral glycoproteins, E1 and E2, are detectable by immunoblots and immunofluorescence for many months but the cells are metabolically inactive and do not synthesize viral proteins. These non-viable cells do not lyse for as long as 2 years.
用辛德毕斯病毒(SFV)感染人白血病造血干细胞系K-562可分为三个阶段:(1)感染后1至4天的早期病毒增殖阶段,在此阶段产生高滴度(10³空斑形成单位/细胞)的传染性病毒,但细胞病变效应最小。通过台盼蓝拒染法,所有细胞看起来都是活的,尽管它们不增殖,并且DNA和细胞蛋白质合成在24小时内降至未感染对照的3%以下;(2)中间阶段,从感染后第5天持续到约第24至30天,在此阶段传染性病毒量降至低水平。在此阶段,病毒蛋白质合成降至无法检测的水平,尽管通过免疫荧光和免疫印迹很容易检测到病毒糖蛋白;然而,衣壳蛋白似乎在感染后21天内降解。细胞数量保持恒定,但用台盼蓝拒染法测定的不增殖细胞的活力无法准确确定;(3)最后一个长期阶段,在此阶段,通过免疫印迹和免疫荧光可在数月内检测到病毒糖蛋白E1和E2,但细胞代谢不活跃,不合成病毒蛋白。这些无活力的细胞长达2年都不会裂解。