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[与慢性感染胡宁病毒的细胞培养物相关的干扰]

[Interference associated with cell cultures chronical by infected with the Junin virus].

作者信息

Help G I, León M E, Coto C E

出版信息

Rev Asoc Argent Microbiol. 1976 May-Aug;8(2):45-53.

PMID:188079
Abstract

Supernatants from Vero cells persistently infected with Junin virus interfered with cytolitic and lethal activities of standard virus. Two Vero cell sublines, chronically infected with Junin virus named VRJ1 and VRJ3, were obtained after prolonged cultivation of cells which survived primary infection. VFJ1 was maintained over a period of 48 days, by biweekly serial transfers while VRJ3, similarly treated, was cultivated for 385 days. One of the characteristics of these cell lines was resistance against superinfection with homologous virus that ordinaily produced CPE and plaques in normal Vero cells; the cells were then considered chronically infected. Supernatants taken at different cell passage level were tested for its interference activity after centrifugation to eliminate floating cells and debris. The degree of CPE intensity caused by inoculation of Vero cells with 10(4), 10(5) or 10(6) TCID 50 of standard virus was markdely deppressed (Figure 1) by coinfection with supernatant from passage 3 of VRJ3 (VRJ1p1), VRJ1p1 supernatant also had interference activity as shown by coinfection with standard virus and expressed by plaque forming inhibition(Table 1). The plaque production of standard virus was inhibited by coinfection with VRJ1p1 supernatant which did not originate plaques when inoculated alone. The interference capacity of VRJ1p6 supernatant was reduced (Table 1) coincidentally with the formation of 55 PFU/ml. Interference activity was neutralized by Junin specific antiserum and inhibited by chloroform treatment. When Vero cells infected with VRJ1p6 supernatant were challenged with standard virus 72 hs later, an inhibition of 98% was achieved (Table 2) in contrast with value of 35% showed in Table 1.

摘要

持续感染胡宁病毒的Vero细胞的上清液干扰了标准病毒的溶细胞和致死活性。在对初次感染后存活的细胞进行长时间培养后,获得了两个长期感染胡宁病毒的Vero细胞亚系,分别命名为VRJ1和VRJ3。通过每两周连续传代,VRJ1维持了48天,而经过类似处理的VRJ3培养了385天。这些细胞系的一个特征是对同源病毒的超感染具有抗性,同源病毒通常会在正常Vero细胞中产生细胞病变效应(CPE)和蚀斑;然后这些细胞被认为是慢性感染。在离心以去除漂浮细胞和碎片后,对不同细胞传代水平获取的上清液进行干扰活性测试。用10⁴、10⁵或10⁶半数组织培养感染剂量(TCID₅₀)的标准病毒接种Vero细胞所引起的CPE强度程度,在与VRJ3传代3(VRJ1p1)的上清液共感染时显著降低(图1),VRJ1p1上清液也具有干扰活性,如与标准病毒共感染所示,并通过蚀斑形成抑制来表示(表1)。标准病毒的蚀斑产生受到与VRJ1p1上清液共感染的抑制,单独接种时该上清液不会产生蚀斑。VRJ1p6上清液的干扰能力随着55蚀斑形成单位/毫升(PFU/ml)的形成而降低(表1)。干扰活性被胡宁特异性抗血清中和,并被氯仿处理抑制。当用VRJ1p6上清液感染的Vero细胞在72小时后用标准病毒攻击时,与表1中显示的35%的值相比,实现了98%的抑制(表2)。

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