Double-Stranded RNA Is Detected by Immunofluorescence Analysis in RNA and DNA Virus Infections, Including Those by Negative-Stranded RNA Viruses
Double-Stranded RNA Is Detected by Immunofluorescence Analysis in RNA and DNA Virus Infections, Including Those by Negative-Stranded RNA Viruses
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DOI：
10.1128/jvi.01299-15
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发表时间：
2015-09-01
影响因子：
5.4
通讯作者：
Lipton, Howard L.
中科院分区：
文献类型：
--
作者：
Son, Kyung-No;Liang, Zhiguo;Lipton, Howard L.
病毒复制的早期生物化学研究表明，大多数病毒产生双链RNA（dsRNA），这是诱导宿主免疫反应所必需的。然而，在2006年报道，dsRNA可以通过免疫荧光抗体染色在双链DNA和正链RNA病毒感染中检测到，但在负链RNA病毒感染中不能检测到。文献中的其他报告似乎支持这些观察结果。这表明负链RNA病毒产生很少的dsRNA，如果有的话，或者更有效的病毒对抗措施来掩盖dsRNA。由于我们对使用dsRNA抗体发现病毒感兴趣，特别是在病理标本中，我们想确定dsRNA的免疫染色在动物病毒感染中的通用性。我们已经检测到在水泡性口炎病毒、麻疹病毒、甲型流感病毒和Nyamanini病毒感染的细胞中dsRNA的原位形成，这些病毒代表来自不同负链RNA病毒家族的病毒。在淋巴细胞性脉络丛脑膜炎病毒（一种双义RNA病毒）和小鼠微小病毒（MVM）（一种单链DNA（ssDNA）细小病毒）感染的细胞中也检测到dsRNA，但未检测到B型肝炎病毒。虽然dsRNA染色主要在细胞质中观察到，但在感染甲型流感病毒、Nyamanini病毒和MVM的细胞的细胞核中也观察到。因此，很可能大多数动物病毒感染产生可通过免疫荧光染色检测的dsRNA种类。在几个未感染的细胞系中诱导的细胞凋亡未能上调dsRNA的形成。重要一点有效的抗病毒宿主免疫反应依赖于识别病毒入侵和完整的先天免疫系统作为第一道防线。双链RNA（dsRNA）是诱导先天免疫所必需的病毒产物，导致I型干扰素（IFN）的产生和数百个IFN刺激基因的激活。目前的研究表明，感染，包括那些由ssDNA病毒和正链和负链RNA病毒，产生的dsRNA可检测的标准免疫荧光染色。虽然dsRNA染色主要在细胞质中观察到，但在一些RNA和DNA病毒感染中也存在核染色。由于宿主dsRNA分子的存在，细胞核不太可能具有针对dsRNA的病原体相关分子模式（PAMP）受体。因此，很可能大多数动物病毒感染产生可通过免疫荧光染色检测的dsRNA种类，这也可证明在病毒发现中是有用的。
Early biochemical studies of viral replication suggested that most viruses produce double-stranded RNA (dsRNA), which is essential for the induction of the host immune response. However, it was reported in 2006 that dsRNA could be detected by immunofluorescence antibody staining in double-stranded DNA and positive-strand RNA virus infections but not in negative-strand RNA virus infections. Other reports in the literature seemed to support these observations. This suggested that negative-strand RNA viruses produce little, if any, dsRNA or that more efficient viral countermeasures to mask dsRNA are mounted. Because of our interest in the use of dsRNA antibodies for virus discovery, particularly in pathological specimens, we wanted to determine how universal immunostaining for dsRNA might be in animal virus infections. We have detected the in situ formation of dsRNA in cells infected with vesicular stomatitis virus, measles virus, influenza A virus, and Nyamanini virus, which represent viruses from different negative-strand RNA virus families. dsRNA was also detected in cells infected with lymphocytic choriomeningitis virus, an ambisense RNA virus, and minute virus of mice (MVM), a single-stranded DNA (ssDNA) parvovirus, but not hepatitis B virus. Although dsRNA staining was primarily observed in the cytoplasm, it was also seen in the nucleus of cells infected with influenza A virus, Nyamanini virus, and MVM. Thus, it is likely that most animal virus infections produce dsRNA species that can be detected by immunofluorescence staining. The apoptosis induced in several uninfected cell lines failed to upregulate dsRNA formation.IMPORTANCEAn effective antiviral host immune response depends on recognition of viral invasion and an intact innate immune system as a first line of defense. Double-stranded RNA (dsRNA) is a viral product essential for the induction of innate immunity, leading to the production of type I interferons (IFNs) and the activation of hundreds of IFN-stimulated genes. The present study demonstrates that infections, including those by ssDNA viruses and positive-and negative-strand RNA viruses, produce dsRNAs detectable by standard immunofluorescence staining. While dsRNA staining was primarily observed in the cytoplasm, nuclear staining was also present in some RNA and DNA virus infections. The nucleus is unlikely to have pathogen-associated molecular pattern (PAMP) receptors for dsRNA because of the presence of host dsRNA molecules. Thus, it is likely that most animal virus infections produce dsRNA species detectable by immunofluorescence staining, which may prove useful in viral discovery as well.