Reiserová L, Kaluzová M, Kaluz S, Willis A C, Závada J, Závodská E, Závadová Z, Ciampor F, Pastorek J, Pastoreková S
Institute of Virology, Slovak Academy of Sciences, Dúbravská cesta 9, Bratislava, 842 46, Slovak Republic.
Virology. 1999 Apr 25;257(1):73-83. doi: 10.1006/viro.1999.9638.
In this study we elucidated the molecular character of MaTu-MX, previously described as an unusual transmissible agent. Amino acid sequencing of peptides generated from a 58-kDa MX-related protein purified from MaTu human carcinoma cells allowed us to identify it as a nucleoprotein (NP) of lymphocytic choriomeningitis virus (LCMV). Northern blot analysis detected LCMV-specific RNAs in MaTu cells. Comparative immunoprecipitations showed cross-reactivity between NP of LCMV strain WE and MX NP. Using RT-PCR, we have cloned MX NP cDNA. According to sequence comparison, MX LCMV is as closely related to both LCMV strains WE and Armstrong as these strains are to one another. Based on this finding we propose that MX is a new strain of LCMV. We also showed that the stability of MX NP in MaTu cells is very high and that the virus is transmissible by cell-to-cell contact or by cell-free extract to human HeLa and monkey Vero cells, but not to human AGS, canine MDCK, mouse NIH 3T3, and hamster CHO cells. Finally, employing MX LCMV NP in immunoprecipitation and solid-phase radioimmunoassay, we found 37.5% prevalence of anti-LCMV antibodies in human sera, suggesting possible horizontal spread of the virus in the human population.
在本研究中,我们阐明了先前被描述为一种不寻常的可传播因子的MaTu-MX的分子特征。对从MaTu人癌细胞中纯化的一种58 kDa的MX相关蛋白产生的肽段进行氨基酸测序,使我们能够将其鉴定为淋巴细胞性脉络丛脑膜炎病毒(LCMV)的核蛋白(NP)。Northern印迹分析在MaTu细胞中检测到LCMV特异性RNA。比较免疫沉淀显示LCMV毒株WE的NP与MX NP之间存在交叉反应。使用RT-PCR,我们克隆了MX NP cDNA。根据序列比较,MX LCMV与LCMV毒株WE和阿姆斯特朗毒株的亲缘关系一样密切,就像这两种毒株彼此之间的关系一样。基于这一发现,我们提出MX是LCMV的一个新毒株。我们还表明,MX NP在MaTu细胞中的稳定性非常高,并且该病毒可通过细胞间接触或无细胞提取物传播给人HeLa细胞和猴Vero细胞,但不能传播给人AGS细胞、犬MDCK细胞、小鼠NIH 3T3细胞和仓鼠CHO细胞。最后,在免疫沉淀和固相放射免疫分析中使用MX LCMV NP,我们发现人血清中抗LCMV抗体的阳性率为37.5%,这表明该病毒可能在人群中存在水平传播。