Schaade L, Kleines M, Krone B, Hausding M, Walter R, Ritter K
Department of Medical Microbiology, University Hospital RWTH Aachen, Germany.
Med Microbiol Immunol. 2000 Sep;189(1):13-8. doi: 10.1007/pl00008252.
Stimulation of Epstein-Barr virus (EBV) genome-positive Burkitt lymphoma cells with the ganglioside IV3NeuAc-nLcOse4Cer leads to the induction of cell differentiation processes and activates the EBV lytic viral cycle. In cells of the Burkitt lymphoma line Raji differential expression of host cell genes was analysed in the early phase (150 min) post stimulation with the ganglioside to display the cell activities that precede the activation of the EBV lytic cycle using the differential display reverse transcription-polymerase chain reaction technique. Multiple fragment cDNAs derived from control cells and ganglioside-stimulated cells were amplified using random primers and displayed via polyacrylamide gel electrophoresis. The expression pattern of 8,400 bands was analysed. Eleven differentially expressed fragment cDNAs were reamplified and identified by nucleotide sequencing. Six of these could be identified as coding for proteins that may take part in virus reactivation and differentiation. The most striking finding was the induction of s-adenosylhomocysteine hydrolase (AHCY) expression. The cellular enzyme AHCY plays an important role in transmethylation reactions controlling the replication of several viruses. Thus. an involvement in EBV replication can be suggested.
用神经节苷脂IV3NeuAc-nLcOse4Cer刺激爱泼斯坦-巴尔病毒(EBV)基因组阳性的伯基特淋巴瘤细胞,可诱导细胞分化过程并激活EBV裂解性病毒周期。在伯基特淋巴瘤细胞系Raji的细胞中,利用差异显示逆转录-聚合酶链反应技术,分析了在用神经节苷脂刺激后的早期阶段(150分钟)宿主细胞基因的差异表达,以展示在EBV裂解周期激活之前的细胞活性。使用随机引物扩增来自对照细胞和神经节苷脂刺激细胞的多个片段cDNA,并通过聚丙烯酰胺凝胶电泳进行展示。分析了8400条带的表达模式。11个差异表达的片段cDNA被重新扩增并通过核苷酸测序进行鉴定。其中6个可被鉴定为编码可能参与病毒再激活和分化的蛋白质。最显著的发现是s-腺苷同型半胱氨酸水解酶(AHCY)表达的诱导。细胞酶AHCY在控制多种病毒复制的转甲基反应中起重要作用。因此,可以推测其参与了EBV复制。