Abstract
BackgroundInfectious bursal disease (IBD) is a highly contagious infectious disease that causes severe immunosuppression and damage to the bursa of Fabricius in chickens. Several proteins involved in IBD virus (IBDV) infection, such as surface immunoglobulin M, integrin, annexin A2 and chicken heat shock protein 90, have been identified. However, the main protein that plays key roles in virus infection has not yet been confirmed.MethodsDF-1 cell line was transfected with the pcDNA-VP2 plasmid and analyzed by immunofluorescence assay. The proteins reacted with VP2 of IBDV in DF-1 cells were pulldown with the monoclonal antibody and identified by mass spectrometry. Heat shock cognate protein 70 (HSC70), one of these proteins, was selected to be investigated in the function in IBDV infection by specific antibody and its inhibitor.ResultsThe DF-1 cell line was transfected with the pcDNA-VP2 plasmid, and expression of IBDV VP2 in DF-1 cells was confirmed by immunofluorescence assays. Heat shock cognate protein 70 (HSC70) was one of the proteins identified by coimmunoprecipitation using a monoclonal antibody (2H11) against VP2 and mass spectrometry analysis. IBDV infection in DF-1 cells was strongly inhibited by both an anti-HSC70 antibody and a HSC70 inhibitor (VER155008).ConclusionThese results suggest that HSC70 may be an essential factor for IBDV infection.
Highlights
Infectious bursal disease (IBD) is a highly contagious infectious disease that causes severe immunosuppression and damage to the bursa of Fabricius in chickens
Many proteins react with IBD virus (IBDV) VP2 The proteins of DF-1 cells transfected with the plasmid pcDNA-VP2 or DF-1 cells transfected with the pcDNA vector were extracted and reacted with a monoclonal antibody against IBDV VP2
Heat shock cognate protein 70 (HSC70) interacts with VP2 according to mass spectrometry analysis Mass spectrometry results (Fig. 2) revealed nearly 45 types of protein molecules able to bind to the VP2 protein
Summary
Infectious bursal disease (IBD) is a highly contagious infectious disease that causes severe immunosuppression and damage to the bursa of Fabricius in chickens. Several proteins involved in IBD virus (IBDV) infection, such as surface immunoglobulin M, integrin, annexin A2 and chicken heat shock protein 90, have been identified. Infectious bursal disease virus (IBDV), a birnavirus belonging to the Birnaviridae family, is a non-enveloped virus that consists of 5 proteins, namely, VP1, VP2, VP3, VP4, and VP5 [1]. As a severe immunosuppressive disease prevalent in chickens, IBDV has resulted in high economic loss in the poultry industry [4]. Hirai et al found that surface immunoglobulin M is a receptor on LSCCBK3 cells [5], and Lin et al showed that chicken HSP90.
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