An Infectious Bursal Disease Virus (IBDV) Reverse Genetics Rescue System and Neutralization Assay in Chicken B Cells.
An Infectious Bursal Disease Virus (IBDV) Reverse Genetics Rescue System and Neutralization Assay in Chicken B Cells.
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DOI：
10.1002/cpz1.639
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发表时间：
2023-01
期刊：
影响因子：
--
通讯作者：
Broadbent, Andrew J.
中科院分区：
文献类型：
--
作者：
Reddy, Vishwanatha R. A. P.;Nazki, Salik;Asfor, Amin;Broadbent, Andrew J.
传染性法氏囊病病毒（IBDV）是家禽业生产力的主要威胁，这是由于发病率、死亡率和免疫抑制，其加剧继发感染并降低疫苗接种程序的效力。IBDV的田间毒株对鸡B细胞具有优选的嗜性，其中大部分存在于法氏囊（BF）中。IBDV对贴壁细胞培养的适应性与改变衣壳蛋白高变区（HVR）中氨基酸的突变有关，其影响免疫原性和毒力。直到最近，这限制了反向遗传学系统在工程分子克隆中的应用，以及体外中和试验在IBDV细胞培养适应株中的应用。在这里，我们描述了救援的IBDV分子克隆含有HVR从不同的领域株，沿着中和试验，以量化抗体反应对获救的病毒，都使用鸡B细胞。这些方法很容易适用于任何具有分子生物学专业知识的实验室，并且不需要获得野生型菌株。版权所有© 2023作者。由Wiley Periodicals LLC出版。 基本方案2：IBDV的鸡B细胞中和试验
Infectious bursal disease virus (IBDV) is a major threat to the productivity of the poultry industry due to morbidity, mortality, and immunosuppression that exacerbates secondary infections and reduces the efficacy of vaccination programs. Field strains of IBDV have a preferred tropism for chicken B cells, the majority of which reside in the bursa of Fabricius (BF). IBDV adaptation to adherent cell culture is associated with mutations altering amino acids in the hypervariable region (HVR) of the capsid protein, which affects immunogenicity and virulence. Until recently, this has limited both the application of reverse genetics systems for engineering molecular clones, and the use of in vitro neutralization assays, to cell‐culture‐adapted strains of IBDV. Here, we describe the rescue of molecular clones of IBDV containing the HVR from diverse field strains, along with a neutralization assay to quantify antibody responses against the rescued viruses, both using chicken B cells. These methods are readily adaptable to any laboratory with molecular biology expertise and negate the need to obtain wild‐type strains. © 2023 The Authors. Current Protocols published by Wiley Periodicals LLC. Basic Protocol 1: A chicken B‐cell rescue system for IBDV Basic Protocol 2: A chicken B‐cell neutralization assay for IBDV
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