Chen F, Zou J Z, di Renzo L, Winberg G, Hu L F, Klein E, Klein G, Ernberg I
Microbiology and Tumor Biology Center, Karolinska Institutet, Stockholm, Sweden.
J Virol. 1995 Jun;69(6):3752-8. doi: 10.1128/JVI.69.6.3752-3758.1995.
Using reverse transcription of whole cellular RNA and nested PCR, we have performed experiments mixing different proportions of Epstein-Barr virus (EBV)-carrying and EBV-negative cells. Based on the results, a method that detects viral transcripts for EBNA-1, EBNA-2, LMP1, and LMP2a from less than one positive cell among 10(5) negative cells was developed. With this method we have shown that the EBV DNA positive cells among small, high-density peripheral blood B-lymphocytes of normal healthy persons express EBNA-1-mRNA but not EBNA-2 or LMP1. A similar EBV expression pattern is found in type I Burkitt lymphoma cells. We suggest that the expression pattern in the lymphoma cells reflects the viral strategy in normal resting B cells and meets the requirements of latent persistence.
我们运用全细胞RNA的逆转录和巢式聚合酶链反应(PCR),对携带爱泼斯坦-巴尔病毒(EBV)和EBV阴性的细胞按不同比例混合进行了实验。基于实验结果,我们开发出一种方法,可从10⁵个阴性细胞中的少于1个阳性细胞中检测出EBNA-1、EBNA-2、LMP1和LMP2a的病毒转录本。通过该方法,我们发现正常健康人的小的、高密度外周血B淋巴细胞中的EBV DNA阳性细胞表达EBNA-1-mRNA,但不表达EBNA-2或LMP1。在I型伯基特淋巴瘤细胞中也发现了类似的EBV表达模式。我们认为淋巴瘤细胞中的这种表达模式反映了病毒在正常静止B细胞中的策略,并且满足潜伏持续存在的要求。