Length-dependent innate antiviral effects of double-stranded RNA in the rainbow trout (Oncorhynchus mykiss) cell line, RTG-2

Poynter, Sarah J.; DeWitte‐Orr, Stephanie J.

doi:10.1016/j.fsi.2015.07.012

Cited by 30 publications

(22 citation statements)

References 40 publications

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“…vig-4 is a VHSV-induced gene; the deduced protein contains tetratricopeptide repeat motifs and shows similarities to the ISG56/IFIT1 family of ISGs ( 9 , 10 ). This study chose vig-4 as a representative transcript indicative of ISG expression because it has been used as a representative ISG in previous studies ( 11 – 14 ) and is upregulated more strongly than other ISGs, such as Mx1 ( 15 ). The dsRNA-induced accumulation of ISG proteins produces a protective antiviral state in rainbow trout cells against various viruses ( 15 , 16 ).…”

Section: Introductionmentioning

confidence: 99%

“…In plasmacytoid dendritic cells only ivt-dsRNA was able to stimulate IFN-α production, poly I:C did not ( 24 ). In rainbow trout cells, ivt-dsRNA induced a faster, stronger IFN1 and IFN2 response compared with poly I:C even when poly I:C was of much longer lengths ( 15 ). In addition, in mice, poly I:C is recognized by MDA5 whereas ivt-dsRNA and v-dsRNA activated RIG-I ( 25 , 28 ).…”

Section: Introductionmentioning

confidence: 99%

“…The current state of research regarding responses to dsRNA largely relies on the use of poly I:C; however, studies of individual receptors are shifting toward ivt-dsRNA, likely for the ease of controlling length ( 25 , 27 , 30 ). Length has been shown to influence the magnitude of immune response in cells and dsRNA receptor types show length requirements and specificities ( 15 , 25 , 27 , 29 ). For example, longer dsRNA molecules have been shown to induce a strong IFN response ( 27 ), and RIG-I has been shown to sense dsRNA molecules under 1,000 bp in length, while MDA5 senses lengths greater than 1,000 bp ( 28 , 30 ).…”

Section: Introductionmentioning

confidence: 99%

“…Rainbow trout were used in this study as a model fish species for their importance in aquaculture and the existing knowledge base of the rainbow trout type I IFN and antiviral response ( 15 , 33 ). In rainbow trout cell lines, ivt-dsRNA or poly I:C induces type I IFN and an antiviral state and similarly whole rainbow trout pretreated with poly I:C also showed decreased susceptibility to a fish virus ( 15 , 34 ).…”

Section: Introductionmentioning

confidence: 99%

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Understanding Viral dsRNA-Mediated Innate Immune Responses at the Cellular Level Using a Rainbow Trout Model

Poynter

DeWitte‐Orr

2018

Front. Immunol.

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Viruses across genome types produce long dsRNA molecules during replication [viral (v-) dsRNA]. dsRNA is a potent signaling molecule and inducer of type I interferon (IFN), leading to the production of interferon-stimulated genes (ISGs), and a protective antiviral state within the cell. Research on dsRNA-induced immune responses has relied heavily on a commercially available, and biologically irrelevant dsRNA, polyinosinic:polycytidylic acid (poly I:C). Alternatively, dsRNA can be produced by in vitro transcription (ivt-) dsRNA, with a defined sequence and length. We hypothesized that ivt-dsRNA, containing legitimate viral sequence and length, would be a more appropriate proxy for v-dsRNA, compared with poly I:C. This is the first study to investigate the effects of v-dsRNA on the innate antiviral response and to compare v-dsRNA to ivt-dsRNA-induced responses in fish cells, specifically rainbow trout. Previously, class A scavenger receptors (SR-As) were found to be surface receptors for poly I:C in rainbow trout cells. In this study, ivt-dsRNA binding was blocked by poly I:C and v-dsRNA, as well as SR-A competitive ligands, suggesting all three dsRNA molecules are recognized by SR-As. Downstream innate antiviral effects were determined by measuring IFN and ISG transcript levels using qRT-PCR and antiviral assays. Similar to what has been shown previously with ivt-dsRNA, v-dsRNA was able to induce IFN and ISG transcript production between 3 and 24 h, and its effects were length dependent (i.e., longer v-dsRNA produced a stronger response). Interestingly, when v-dsRNA and ivt-dsRNA were length and sequence matched both molecules induced statistically similar IFN and ISG transcript levels, which resulted in similar antiviral states against two aquatic viruses. To pursue sequence effects further, three ivt-dsRNA molecules of the same length but different sequences (including host and viral sequences) were tested for their ability to induce IFN/ISG transcripts and an antiviral state. All three induced responses similarly. This study is the first of its kind to look at the effects v-dsRNA in fish cells as well as to compare ivt-dsRNA to v-dsRNA, and suggests that ivt-dsRNA may be a good surrogate for v-dsRNA in the study of dsRNA-induced responses and potential future antiviral therapies.

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Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Understanding Viral dsRNA-Mediated Innate Immune Responses at the Cellular Level Using a Rainbow Trout Model

Poynter

DeWitte‐Orr

2018

Front. Immunol.

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“…Type I IFN (Poynter and DeWitte-Orr, 2015), may mimic extracellular viral dsRNA that is 99 released from lysed infected cells. The objectives of this study were to first generate and 100 characterize novel X. laevis skin epithelial cell lines and then use these newly generated frog skin 101…”

mentioning

confidence: 99%

Xela DS2 and Xela VS2: two novel skin epithelial-like cell lines from adult African clawed frog (Xenopus laevis) and their response to an extracellular viral dsRNA analogue

Bui-Marinos

Varga

et al. 2020

Preprint

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18The skin epithelial layer acts as an important immunological barrier against pathogens 19 and is capable of recognizing and responding to pathogen associated molecular patterns 20 (PAMPs) in human and mouse models. Although presumed, it is unknown whether amphibian 21 skin epithelial cells exhibit the ability to respond to PAMPs such as viral double-stranded RNA 22(dsRNA). To address this, two cell lines from the dorsal skin (Xela DS2) and ventral skin (Xela 23 VS2) of the African clawed frog (Xenopus laevis) were established. Xela DS2 and Xela VS2 24 cells have an epithelial-like morphology, express genes associated with epithelial cells, and lack 25 senescence-associated beta-galactosidase activity. Cells grow optimally in 70% Leibovitz's L-15 26 medium supplemented with 15% fetal bovine serum at 26°C. Upon treatment with poly(I:C), a 27 synthetic viral dsRNA analogue and known type I interferon inducer, Xela DS2 and Xela VS2 28 exhibit marked upregulation of key pro-inflammatory and antiviral transcripts suggesting frog 29 epithelial cells participate in the recognition of extracellular viral dsRNA and production of local 30 inflammatory signals; similar to human and mouse models. Currently, these are the only known 31Xenopus laevis skin epithelial-like cell lines and will be important for future research in 32 amphibian epithelial cell biology, initial host-pathogen interactions, and rapid screening of the 33 effects of environmental stressors, including contaminants, on frog skin epithelial cells. 34 35

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Invitromatics, invitrome, and invitroomics: introduction of three new terms for in vitro biology and illustration of their use with the cell lines from rainbow trout

Bols

Pham

Dayeh

et al. 2017

In Vitro Cell.Dev.Biol.-Animal

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The literature on cell lines that have been developed from rainbow trout (RT) (Oncorhynchus mykiss) is reviewed to illustrate three new terms: invitromatics, invitrome, and invitroomics. Invitromatics is defined as the history, development, characterization, engineering, storage, and sharing of cell lines. RT invitromatics differs from invitromatics for humans and other mammals in several ways. Nearly all the RT cell lines have developed through spontaneous immortalization. No RT cell line undergoes senescence and can be described as being finite, whereas many human cell lines undergo senescence and are finite. RT cell lines are routinely grown at 18-22°C in free gas exchange with air in basal media developed for mammalian cells together with a supplement of fetal bovine serum. An invitrome is defined as the grouping of cell lines around a theme or category. The broad theme in this article is all the cell lines that have ever been created from O. mykiss, or in other words, the RT invitrome. The RT invitrome consists of approximately 55 cell lines. These cell lines can also be categorized on the basis of their storage and availability. A curated invitrome constitutes all the cell lines in a repository and for RT consists of 11 cell lines. These consist of epithelial cell lines, such as RTgill-W1, and fibroblast cell lines, such as RTG-2. RTG-2 can be purchased from a scientific company and constitutes the commercial RT invitrome. Cell lines that are exchanged between researchers are termed the informally shared invitrome and for RT consists of over 35 cell lines. Among these is the monocyte/macrophage cell line, RTS11. Cell lines whose existence is in doubt are termed the zombie invitrome, and for RT, approximately 12 cell lines are zombies. Invitroomics is the application of cell lines to a scientific problem or discipline. This is illustrated with the use of the RT invitrome in virology. Of the RT invitrome, RTG-2 was the most commonly used cell line to isolate viruses. Fifteen families of viruses were studied with RT invitrome. RT cell lines were best able to support replication of viruses from the Herpesviridae, Iridoviridae, Birnaviridae, Togaviridae, and Rhabdoviridae families.

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Length-dependent innate antiviral effects of double-stranded RNA in the rainbow trout (Oncorhynchus mykiss) cell line, RTG-2

Cited by 30 publications

References 40 publications

Understanding Viral dsRNA-Mediated Innate Immune Responses at the Cellular Level Using a Rainbow Trout Model

Understanding Viral dsRNA-Mediated Innate Immune Responses at the Cellular Level Using a Rainbow Trout Model

Xela DS2 and Xela VS2: two novel skin epithelial-like cell lines from adult African clawed frog (Xenopus laevis) and their response to an extracellular viral dsRNA analogue

Invitromatics, invitrome, and invitroomics: introduction of three new terms for in vitro biology and illustration of their use with the cell lines from rainbow trout

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