School of Cancer Sciences, College of Medical and Dental Sciences, University of Birmingham, B15 2TT, UK.
J Virol Methods. 2013 Oct;193(1):251-9. doi: 10.1016/j.jviromet.2013.06.001. Epub 2013 Jun 10.
Kaposi's sarcoma-associated herpesvirus (KSHV) is the aetiologic agent of Kaposi's sarcoma (KS), a tumour of endothelial cell origin. The study of KS development was aided by the generation of a recombinant GFP (latent)/RFP (lytic)-expressing KSHV (rKSHV.219) by Vieira and O'Hearn (2004). In this study the first data characterising primary endothelial cell infection and transmission with this virus is presented. Infection was predominantly latent and the percentage of GFP-positive cells increased over time. Neither horizontal transmission of infection, nor cellular proliferation, explained this increase. Analysis of latency-associated nuclear antigen (LANA-1) expression revealed that a threshold level of infection was required for GFP expression early post infection. At later time points GFP correlated more closely with LANA-1 expression, likely due to the accumulation of GFP over time. This study provides methodological guidance for the use of rKSHV.21. In addition, it highlights potential problems associated with the use of fluorescent proteins as markers of viral infection.
卡波氏肉瘤相关疱疹病毒（KSHV）是卡波氏肉瘤（KS）的病原体，KS 是一种起源于内皮细胞的肿瘤。通过 Vieira 和 O'Hearn（2004）生成表达 GFP（潜伏）/RFP（裂解）的重组 KSHV（rKSHV.219），KS 的发展研究得到了帮助。在这项研究中，首次提出了该病毒对原发性内皮细胞感染和传播的特征数据。感染主要是潜伏性的，GFP 阳性细胞的百分比随时间增加。水平传播感染和细胞增殖都不能解释这种增加。潜伏相关核抗原（LANA-1）表达的分析表明，在感染后早期 GFP 表达需要达到感染的阈值水平。在稍后的时间点，GFP 与 LANA-1 的表达更为密切相关，这可能是由于 GFP 随时间的积累。本研究为 rKSHV.21 的使用提供了方法学指导。此外，它还强调了使用荧光蛋白作为病毒感染标志物可能存在的问题。
