Dulwich Katherine L, Giotis Efstathios S, Gray Alice, Nair Venugopal, Skinner Michael A, Broadbent Andrew J
The Pirbright Institute, Ash Road, Woking, GU24 0NF, UK.
Section of Virology, Faculty of Medicine, Imperial College London, St. Mary's Campus, Norfolk Place, London W2 1PG, UK.
J Gen Virol. 2017 Dec;98(12):2918-2930. doi: 10.1099/jgv.0.000979. Epub 2017 Nov 20.
Infectious bursal disease virus (IBDV) belongs to the family Birnaviridae and is economically important to the poultry industry worldwide. IBDV infects B cells in the bursa of Fabricius (BF), causing immunosuppression and morbidity in young chickens. In addition to strains that cause classical Gumboro disease, the so-called 'very virulent' (vv) strain, also in circulation, causes more severe disease and increased mortality. IBDV has traditionally been controlled through the use of live attenuated vaccines, with attenuation resulting from serial passage in non-lymphoid cells. However, the factors that contribute to the vv or attenuated phenotypes are poorly understood. In order to address this, we aimed to investigate host cell-IBDV interactions using a recently described chicken primary B-cell model, where chicken B cells are harvested from the BF and cultured ex vivo in the presence of chicken CD40L. We demonstrated that these cells could support the replication of IBDV when infected ex vivo in the laboratory. Furthermore, we evaluated the gene expression profiles of B cells infected with an attenuated strain (D78) and a very virulent strain (UK661) by microarray. We found that key genes involved in B-cell activation and signalling (TNFSF13B, CD72 and GRAP) were down-regulated following infection relative to mock, which we speculate could contribute to IBDV-mediated immunosuppression. Moreover, cells responded to infection by expressing antiviral type I IFNs and IFN-stimulated genes, but the induction was far less pronounced upon infection with UK661, which we speculate could contribute to its virulence.
传染性法氏囊病病毒(IBDV)属于双RNA病毒科,对全球家禽业具有重要经济意义。IBDV感染法氏囊(BF)中的B细胞,导致幼鸡免疫抑制和发病。除了引起经典甘博罗病的毒株外,所谓的“超强毒”(vv)毒株也在传播,会导致更严重的疾病和更高的死亡率。传统上,IBDV是通过使用减毒活疫苗来控制的,减毒是通过在非淋巴细胞中连续传代实现的。然而,导致vv或减毒表型的因素尚不清楚。为了解决这个问题,我们旨在使用最近描述的鸡原代B细胞模型研究宿主细胞与IBDV的相互作用,该模型中从BF收获鸡B细胞,并在鸡CD40L存在的情况下进行体外培养。我们证明,这些细胞在实验室中体外感染时能够支持IBDV的复制。此外,我们通过微阵列评估了感染减毒株(D78)和超强毒株(UK661)的B细胞的基因表达谱。我们发现,与未感染对照组相比,感染后参与B细胞活化和信号传导的关键基因(TNFSF13B、CD72和GRAP)表达下调,我们推测这可能导致IBDV介导的免疫抑制。此外,细胞通过表达抗病毒I型干扰素和干扰素刺激基因对感染做出反应,但感染UK661后这种诱导作用远不明显,我们推测这可能与其毒力有关。