Type II Grass Carp Reovirus Infects Leukocytes but Not Erythrocytes and Thrombocytes in Grass Carp (Ctenopharyngodon idella)

Yang, Ling; Su, Jian

doi:10.3390/v13050870

Cited by 9 publications

(5 citation statements)

References 48 publications

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“…To examine mRNA distribution in blood cells, grass carp blood was collected by caudal venipuncture with a syringe that contain 5–10% 0.5‐M EDTA (pH 8.0). The peripheral blood erythrocytes, leukocytes, and thrombocytes were isolated using a fish peripheral blood platelet separation kit (P1740, Solarbio, Beijing, China) according to manufacturer instructions (Yang & Su, 2021) ( n = 4). Total RNA was isolated and reverse transcribed for qRT–PCR.…”

Section: Methodsmentioning

confidence: 99%

Cyprinid‐specific duplicated membrane TLR5 senses dsRNA as functional homodimeric receptors

et al. 2022

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Membrane‐embedded Toll‐like receptor 5 (TLR5) functions as a homodimer to detect bacterial flagellin. Cyprinid grass carp (Ctenopharyngodon idella) encodes two TLR5 genes, CiTLR5a and CiTLR5b. Here, we show that cyprinid TLR5a and TLR5b homodimers unexpectedly bind the dsRNA analog poly(I:C) and regulate interferon (IFN) response in early endosomes and lysosomes. Although TLR5 homodimers also bind flagellin, an immune response to flagellin is only triggered by TLR5a/b heterodimer. Moreover, we demonstrate that two TLR5 paralogs have opposite effects on antiviral response: CiTLR5a slightly promotes and powerfully maintains, whereas CiTLR5b remarkably inhibits virus replication. We show that the ectodomain of CiTLR5 is required for dsRNA‐induced IFN signaling, and we map the key poly(I:C) binding sites to G240 for CiTLR5a and to N547 for CiTLR5b. Furthermore, we reveal that differential N‐glycosylation of CiTLR5a/b affects dsRNA‐IFN signaling but has no role in flagellin‐mediated NF‐κB induction, with paralog‐specific roles for CiTLR5a‐T101 and corresponding CiTLR5b‐I99. Moreover, we provide evidence that the ability to sense dsRNA represents a neofunctionalization specific for membrane‐bound TLR5 in cyprinid, bridging viral and bacterial immune responses.

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Section: Methodsmentioning

confidence: 99%

Cyprinid‐specific duplicated membrane TLR5 senses dsRNA as functional homodimeric receptors

et al. 2022

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“…GCRV-II easily breaks through the thin monolayer flattened epithelial cells in branchial lamella and enters the blood circulation, which is fast in gill, thus, virus is transported to other tissues, so virus can be detectable at 5 min by semi-qRT-PCR but detectable at 45 min by WB. Previous study indicated that GCRV-II can infect leukocytes [22]. Thus, GCRV-II can pass the blood-brain barrier.…”

Section: Discussionmentioning

confidence: 96%

“…Nostril, olfactory bulb, olfactory tract, and brain tissues of immersion infected grass carp were fixed with 2.5% glutaraldehyde in a 0.1 M phosphate buffer (pH 7.2) at 4°C for 24 h to observe the virus particles. Ultrathin sections were prepared as previously described [22]. Images were viewed on an HT-7700 TEM (Hitachi, Japan).…”

Section: Transmission Electron Microscopymentioning

confidence: 99%

Type II Grass Carp Reovirus Rapidly Invades Grass Carp (<em>Ctenopharyngodon idella</em>) via Nostril-Olfactory System -Brain Axis, Gill, and Skin on Head

Zhu¹,

Qiao²,

Hu³

et al. 2023

Preprint

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Type II grass carp reovirus (GCRV-II) with high pathogenicity and infectivity causes severe hemorrhagic disease, which leads to extensive death in the grass carp and black carp aquaculture. However, the early invasion portal remains unclear. In this study, we explored the invasion portal, time, and pathway of GCRV-II by immersion infection in grass carp. Through the detection of the infected grass carp external body surface tissues, most of them could be detectable to carry GCRV-II within 45 minutes except for the skin covered by scales. Further shortening the duration of infection, we proved that GCRV-II rapidly invades through the nostril (especially), gill, and skin on head at only 5 min post immersion, rather than merely adhesion. Subsequently, visual localization investigations of GCRV-II were conducted on the nostril, olfactory system (olfactory bulb and olfactory tract), and brain by immunofluorescence microscopy and transmission electron microscopy. We found that few viruses were located in the nostril at 5 min post immersion infection, while significantly increased viruses were distributed in all these tissues at 45 min. Furthermore, semi-qRT-PCR and western blotting results of different infection time confirmed that GCRV-II invades grass carp via nostril-olfactory system-brain axis and then unfolds viral replication. These results revealed the infection mechanism of GCRV-II on the invasion portal, time, and pathway in grass carp. This study aims to understand the invasion mode of GCRV-II in grass carp, which provides theoretical support for the prevention and control strategies of hemorrhagic disease.

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“…GCRV-II easily breaks through the thin monolayer flattened epithelial cells in branchial lamella and enters blood circulation, which is fast in gills, thus, the virus is transported to other tissues; hence, the virus can be detectable at 5 min via semi-qRT-PCR but detectable at 45 min via WB. Previous study has indicated that GCRV-II can infect leukocytes [ 24 ]. Thus, GCRV-II can pass the blood–brain barrier.…”

Section: Discussionmentioning

confidence: 99%

“…The nostril, olfactory bulb, olfactory tract, and brain tissues of immersion infected grass carp were fixed with 2.5% glutaraldehyde in a 0.1 M phosphate buffer (pH 7.2) at 4 °C for 24 h to observe the virus particles. Ultrathin sections were prepared as previously described [ 24 ]. Images were viewed on an HT-7700 TEM (Hitachi, Japan).…”

Section: Methodsmentioning

confidence: 99%

Type II Grass Carp Reovirus Rapidly Invades Grass Carp (Ctenopharyngodon idella) via Nostril–Olfactory System–Brain Axis, Gill, and Skin on Head

Zhu

Qiao

et al. 2023

Viruses

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Type II grass carp reovirus (GCRV-II) with high pathogenicity and infectivity causes severe hemorrhagic disease, which leads to extensive death in the grass carp and black carp aquaculture. However, the early invasion portal remains unclear. In this study, we explored the invasion portal, time, and pathway of GCRV-II by immersion infection in grass carp. Through the detection of the infected grass carp external body surface tissues, most of them could be detected to carry GCRV-II within 45 min except for the skin covered by scales. Further shortening the duration of infection, we proved that GCRV-II rapidly invades through the nostril (especially), gill, and skin on head at only 5 min post-immersion, rather than merely by adhesion. Subsequently, visual localization investigations of GCRV-II were conducted on the nostril, olfactory system (olfactory bulb and olfactory tract), and brain via immunofluorescence microscopy and transmission electron microscopy. We found that few viruses were located in the nostril at 5 min post-immersion infection, while a significantly increased quantity of viruses were distributed in all of the examined tissues at 45 min. Furthermore, the semi-qRT-PCR and Western blotting results of different infection times confirmed that GCRV-II invades grass carp via the nostril–olfactory system–brain axis and then viral replication unfolds. These results revealed the infection mechanism of GCRV-II in terms of the invasion portal, time, and pathway in grass carp. This study aims to understand the invasion mode of GCRV-II in grass carp, thus providing theoretical support for the prevention and control strategies of hemorrhagic disease.

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Type II Grass Carp Reovirus Infects Leukocytes but Not Erythrocytes and Thrombocytes in Grass Carp (Ctenopharyngodon idella)

Cited by 9 publications

References 48 publications

Cyprinid‐specific duplicated membrane TLR5 senses dsRNA as functional homodimeric receptors

Cyprinid‐specific duplicated membrane TLR5 senses dsRNA as functional homodimeric receptors

Type II Grass Carp Reovirus Rapidly Invades Grass Carp (<em>Ctenopharyngodon idella</em>) via Nostril-Olfactory System -Brain Axis, Gill, and Skin on Head

Type II Grass Carp Reovirus Rapidly Invades Grass Carp (Ctenopharyngodon idella) via Nostril–Olfactory System–Brain Axis, Gill, and Skin on Head

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