Isolation and characterization of an interferon-resistant cell line deficient in the induction of (2'-5')oligoadenylate synthetase activity
Isolation and characterization of an interferon-resistant cell line deficient in the induction of (2'-5')oligoadenylate synthetase activity
复制标题
缺乏 (2-5) 寡腺苷酸合成酶活性诱导的干扰素抗性细胞系的分离和表征
DOI：
10.1128/mcb.3.10.1759-1765.1983
复制
发表时间：
1983
影响因子：
5.3
通讯作者：
M. Bakhanashvili
中科院分区：
文献类型：
--
作者：
S. Salzberg;D. Wreschner;F. Oberman;A. Panet;M. Bakhanashvili
为了筛选对干扰素（IFN）具有不同敏感性的细胞，对NIH 3T3小鼠成纤维细胞进行了亚克隆，并检测了它们对IFN治疗的反应。在测试的30个克隆中，2个似乎对IFN具有相对抗性，因为即使在每毫升1000 U IFN存在的情况下，水疱性口炎病毒和孟gov病毒的复制都没有受到抑制。对一个耐药克隆（A10）和一个敏感克隆（A5）进行了进一步分析。在两个克隆中，小鼠白血病病毒的复制同样受到IFN的抑制，这表明在耐药克隆中存在IFN的功能受体。通过（2’-5’）寡聚腺苷酸（2- 5a）放射结合实验，我们可以证明两个克隆都含有RNase L蛋白。此外，这种酶似乎是活跃的，因为在外源2-5A引入细胞后，蛋白质合成速率明显降低。我们还分析了2-5A途径中另一种酶的活性，即2-5A合成酶。在敏感细胞（A5）中，酶活性的诱导与使用的IFN浓度成正比，在未处理细胞的背景下达到最大10倍以上的增加。然而，当使用相同剂量的IFN时，在抗性细胞（A10）中几乎没有观察到任何对基础活性的诱导。因此，在A10细胞中，IFN缺乏对2-5A合成酶活性的诱导可能至少是它们对IFN治疗相对耐药的部分原因。
To screen for cells with different sensitivities to interferon (IFN), NIH 3T3 mouse fibroblasts were subcloned and examined for their response to IFN treatment. Of 30 clones tested, 2 appeared to be relatively resistant to IFN, since the replication of both vesicular stomatitis virus and mengovirus was not inhibited, even in the presence of 1,000 U of IFN per ml. One resistant (A10) and one sensitive (A5) clone were further analyzed. In both clones, murine leukemia virus replication was equally inhibited by IFN, indicating the presence of functional receptors for IFN in the resistant clone. Using the (2'-5')oligoadenylate (2-5A) radiobinding assay, we could demonstrate that both clones contained the RNase L protein. Furthermore, this enzyme appears to be active, since a similar reduction in the rate of protein synthesis was evident after the introduction of exogenous 2-5A to the cells. We also analyzed the activity of another enzyme in the 2-5A pathway, namely, 2-5A synthetase. In the sensitive cells (A5), the induction of enzyme activity was proportional to the IFN concentration used, reaching a maximum of more than a 10-fold increase over the background of untreated cells. However, little if any induction over the basal activity was observed in the resistant cells (A10) when similar doses of IFN were used. It is thus probable that the lack of induction of 2-5A synthetase activity by IFN in A10 cells is at least partly responsible for their relative resistance to IFN treatment.
干扰素对呼肠孤病毒的作用：呼肠孤病毒感染后小鼠 L929 细胞中干扰素诱导的蛋白激酶的激活。
影响因子：
3.7
作者：
Gupta,SL;Holmes,SL;Mehra,LL
通讯作者：
Mehra,LL