scholarly journals Inhibition of infectious bursal disease virus infection by artificial microRNAs targeting chicken heat-shock protein 90

2012 ◽  
Vol 93 (4) ◽  
pp. 876-879 ◽  
Author(s):  
Weifeng Yuan ◽  
Xinyu Zhang ◽  
Xiaoli Xia ◽  
Huaichang Sun
Keyword(s):  
Heat Shock ◽  
Heat Shock Protein ◽  
Infectious Bursal Disease Virus ◽  
Heat Shock Protein 90 ◽  
Infectious Bursal Disease ◽  
Cellular Receptor ◽  
Receptor Complex ◽  
Rt Pcr ◽  
Transfected Cells ◽  
Bursal Disease

Infectious bursal disease virus (IBDV) causes an important disease in young chickens. Chicken heat-shock protein 90 (cHsp90) has been shown to be a functional component of the cellular receptor complex for IBDV infection. This study demonstrates the inhibitory effect of vector-expressed anti-cHsp90α microRNA (miRNA) on IBDV infection. The reporter vectors pcHsp90α-EGFP and pcHsp90β-EGFP were constructed to facilitate effective miRNA selection. Two anti-cHsp90α and one anti-cHsp90β miRNA-expression vectors were constructed for a stable transfection study. Poly(A)-tailed RT-PCR detected sequence-specific miRNA transcription in transfected cells. Semiquantitative RT-PCR showed inhibition of cHsp90 transcription in transfected cells. A virus-titration assay showed that the anti-cHsp90α miRNA, but not the anti-cHsp90β miRNA, had inhibitory effects on IBDV infection. These results suggest that cHsp90α is a functional component of the cellular receptor complex for IBDV infection, and that anti-cHsp90α miRNA could be used as an anti-IBDV reagent.

scholarly journals Chicken Heat Shock Protein 90 Is a Component of the Putative Cellular Receptor Complex of Infectious Bursal Disease Virus

2007 ◽  
Vol 81 (16) ◽  
pp. 8730-8741 ◽  
Author(s):  
Ta-Wei Lin ◽  
Chi-Wen Lo ◽  
Su-Yuan Lai ◽  
Ruey-Jane Fan ◽  
Chao-Jung Lo ◽  
...  
Keyword(s):  
Heat Shock ◽  
Heat Shock Protein ◽  
Infectious Bursal Disease Virus ◽  
Heat Shock Protein 90 ◽  
Infectious Bursal Disease ◽  
Cellular Receptor ◽  
Dependent Manner ◽  
Receptor Complex ◽  
Bursal Disease ◽  
Dose Dependent

ABSTRACT Infectious bursal disease virus (IBDV) causes a highly contagious disease in young chicks and leads to significant economic losses in the poultry industry. The capsid protein VP2 of IBDV plays an important role in virus binding and cell recognition. VP2 forms a subviral particle (SVP) with immunogenicity similar to that of the IBDV capsid. In the present study, we first showed that SVP could inhibit IBDV infection to an IBDV-susceptible cell line, DF-1 cells, in a dose-dependent manner. Second, the localizations of the SVP on the surface of DF-1 cells were confirmed by fluorescence microscopy, and the specific binding of the SVP to DF-1 cells occurred in a dose-dependent manner. Furthermore, the attachment of SVP to DF-1 cells was inhibited by an SVP-induced neutralizing monoclonal antibody against IBDV but not by denatured-VP2-induced polyclonal antibodies. Third, the cellular factors in DF-1 cells involved in the attachment of SVP were purified by affinity chromatography using SVP bound on the immobilized Ni2+ ions. A dominant factor was identified as being chicken heat shock protein 90 (Hsp90) (cHsp90) by mass spectrometry. Results of biotinylation experiments and indirect fluorescence assays indicated that cHsp90 is located on the surface of DF-1 cells. Virus overlay protein binding assays and far-Western assays also concluded that cHsp90 interacts with IBDV and SVP, respectively. Finally, both Hsp90 and anti-Hsp90 can inhibit the infection of DF-1 cells by IBDV. Taken together, for the first time, our results suggest that cHsp90 is part of the putative cellular receptor complex essential for IBDV entry into DF-1 cells.

scholarly journals Chicken Heat Shock Protein 70 Is an Essential Host Protein for Infectious Bursal Disease Virus Infection In Vitro

Pathogens ◽  
2021 ◽  
Vol 10 (6) ◽  
pp. 664
Author(s):  
Yufang Meng ◽  
Xiaoxue Yu ◽  
Chunxue You ◽  
Wenjuan Zhang ◽  
Yingfeng Sun ◽  
...  
Keyword(s):  
Heat Shock ◽  
Heat Shock Protein ◽  
Disease Virus ◽  
Heat Shock Protein 70 ◽  
Infectious Bursal Disease Virus ◽  
Host Protein ◽  
Infectious Bursal Disease ◽  
Economic Losses ◽  
Small Interfering Rnas ◽  
Bursal Disease

Infectious bursal disease virus (IBDV) infection causes pathogenicity and mortality in chickens, leading to huge economic losses in the poultry industry worldwide. Studies of host-virus interaction can help us to better understand the viral pathogenicity. As a highly conservative host factor, heat shock protein 70 (Hsp70) is observed to be involved in numerous viral infections. However, there is little information about the role of chicken Hsp70 (cHsp70) in IBDV infection. In the present study, the increased expression of cHsp70 was observed during IBDV-infected DF-1 cells. Further studies revealed that Hsp70 had similar locations with the viral double-stranded RNA (dsRNA), and the result of pull-down assay showed the direct interaction between cHsp70 with dsRNA, viral proteins (vp)2 and 3, indicating that maybe cHsp70 participates in the formation of the replication and transcription complex. Furthermore, overexpression of cHsp70 promoted IBDV production and knockdown of cHsp70 using small interfering RNAs (siRNA) and reducedviral production, implying the necessity of cHsp70 in IBDV infection. These results reveal that cHsp70 is essential for IBDV infection in DF-1 cells, suggesting that targeting cHsp70 may be applied as an antiviral strategy.

A Practical Tissue Sampling Method Using Ordinary Paper for Molecular Detection of Infectious Bursal Disease Virus RNA by RT-PCR

2006 ◽  
Vol 50 (4) ◽  
pp. 556-560 ◽  
Author(s):  
Min Thein Maw ◽  
Tsuyoshi Yamaguchi ◽  
Christopher J. Kasanga ◽  
Kaori Terasaki ◽  
Hideto Fukushi
Keyword(s):  
Disease Virus ◽  
Infectious Bursal Disease Virus ◽  
Sampling Method ◽  
Molecular Detection ◽  
Infectious Bursal Disease ◽  
Tissue Sampling ◽  
Rt Pcr ◽  
Virus Rna ◽  
Bursal Disease

Some characteristics of a cellular receptor for virulent infectious bursal disease virus by using flow cytometry

1998 ◽  
Vol 143 (12) ◽  
pp. 2327-2341 ◽  
Author(s):  
M. Ogawa ◽  
T. Yamaguchi ◽  
A. Setiyono ◽  
T. Ho ◽  
H. Matsuda ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Disease Virus ◽  
Infectious Bursal Disease Virus ◽  
Infectious Bursal Disease ◽  
Cellular Receptor ◽  
Bursal Disease

scholarly journals Effective inhibition of infectious bursal disease virus replication by recombinant avian adeno-associated virus-delivered microRNAs

2009 ◽  
Vol 90 (6) ◽  
pp. 1417-1422 ◽  
Author(s):  
Yongjuan Wang ◽  
Huaichang Sun ◽  
Pengpeng Shen ◽  
Xinyu Zhang ◽  
Xiaoli Xia
Keyword(s):  
Disease Virus ◽  
Infectious Bursal Disease Virus ◽  
Viral Vector ◽  
Infectious Bursal Disease ◽  
Inhibitory Effects ◽  
Rt Pcr ◽  
Adeno Associated Virus ◽  
Rnai Technology ◽  
Bursal Disease ◽  
In Cells

RNA interference (RNAi) is a novel antiviral strategy against a variety of virus infections. Infectious bursal disease virus (IBDV) causes an economically important disease in young chickens. This study demonstrated efficient inhibition of IBDV replication by recombinant avian adeno-associated virus (rAAAV)-delivered anti-VP1 and anti-VP2 microRNAs (miRNAs). In the viral vector-transduced cells, sequence-specific miRNA expression was detected by poly(A)-tailed RT-PCR. Reporter assays using a pVP2-EGFP vector showed significant and long-lasting inhibition of VP2–EGFP expression in cells transduced with anti-VP2 miRNA-expressing rAAAV-RFPmiVP2E, but not with the control miRNA-expressing rAAAV-RFPmiVP2con or anti-VP1 miRNA-expressing rAAAV-RFPmiVP1. Semi-quantitative RT-PCR and/or virus titration assays showed a significant inhibitory effect on homologous IBDV replication in cells transduced with rAAAV-RFPmiVP1 or rAAAV-RFPmiVP2E. For two heterologous IBDV isolates, transduction with rAAAV-RFPmiVP1 led to slightly weaker but similar inhibitory effects, whereas transduction with rAAAV-RFPmiVP2E resulted in significantly weaker and different inhibitory effects. These results suggest that rAAAV could act as an efficient vector for miRNA delivery into avian cells and that VP1 is the more suitable target for interfering with IBDV replication using RNAi technology.

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