Kinetics of murine gammaherpesvirus 68 gene expression following infection of murine cells in culture and in mice.
Kinetics of murine gammaherpesvirus 68 gene expression following infection of murine cells in culture and in mice.
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培养物和小鼠体内的鼠科细胞感染后鼠伽马疱疹病毒 68 基因表达的动力学。
DOI：
10.1128/jvi.75.11.4955-4963.2001
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发表时间：
2001
影响因子：
5.4
通讯作者：
Cardin,RD
中科院分区：
文献类型：
--
作者：
Rochford,R;Lutzke,ML;Alfinito,RS;Clavo,A;Cardin,RD
研究γ疱疹病毒感染的发病机制的模型系统是用鼠γ疱疹病毒68（MHV-68）感染小鼠。为了确定感染的动力学，我们开发了一种RNA酶保护试验，以定量裂解（K3、Rta、M8、DNA聚合酶[DNA pol]和gB）和候选潜伏期（M2、M3、M9、M11、ORF 73和ORF 74）基因的基因表达。所有候选潜伏基因在裂解感染3 T3细胞期间表达。在感染3 T3细胞后观察到四种动力学类型的转录物：立即早期（K3、Rta、M8和ORF 73）、早期（DNA pol）、早期晚期（M3、M11和ORF 74）和晚期（M2、M9和gB）。为了评估体内病毒基因表达的动力学，从MHV-68感染的小鼠收获肺、脾和纵隔淋巴结（MLN）。所有的成绩单之间的3和6天感染后（dpi）在肺中表达。在脾脏中，当潜伏期建立时，K3、M3、M8和M9转录本在10和16 dpi之间表达。从2到16 dpi在MLN中检测到K3、M3、M8、M9和M11转录本。这是首次证实MHV-68基因在MLN中表达。重要的是，我们的数据表明，MHV-68在不同的感染部位具有不同的基因表达动力学。此外，我们证明，K3，一个基因最近被证明编码一种蛋白质，下调细胞表面的主要组织相容性复合体I类，在潜伏期，这认为在潜伏感染期间的免疫逃避K3的作用。
A model system to study the pathogenesis of gammaherpesvirus infections is the infection of mice with murine gammaherpesvirus 68 (MHV-68). To define the kinetics of infection, we developed an RNase protection assay to quantitate gene expression from lytic (K3, Rta, M8, DNA polymerase [DNA pol], and gB) and candidate latency (M2, M3, M9, M11, ORF73, and ORF74) genes. All candidate latency genes were expressed during lytic infection of 3T3 cells. Four kinetic classes of transcripts were observed following infection of 3T3 cells: immediate-early (K3, Rta, M8, and ORF73), early (DNA pol), early-late (M3, M11, and ORF74), and late (M2, M9, and gB). To assess the kinetics of viral gene expression in vivo, lungs, spleens, and mediastinal lymph nodes (MLN) were harvested from MHV-68-infected mice. All transcripts were expressed between 3 and 6 days postinfection (dpi) in the lungs. In the spleen, K3, M3, M8, and M9 transcripts were expressed between 10 and 16 dpi when latency is established. The K3, M3, M8, M9, and M11 transcripts were detected in the MLN from 2 through 16 dpi. This is the first demonstration of MHV-68 gene expression in the MLN. Importantly, our data showed that MHV-68 has different kinetics of gene expression at different sites of infection. Furthermore, we demonstrated that K3, a gene recently shown to encode a protein that downregulates major histocompatibility complex class I on the surface of cells, is expressed during latency, which argues for a role of K3 in immune evasion during latent infection.