Identification of a potential neutralizing linear epitope of hemagglutinin-neuraminidase in Newcastle disease virus.
Identification of a potential neutralizing linear epitope of hemagglutinin-neuraminidase in Newcastle disease virus.
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DOI：
10.1186/s12985-020-01483-y
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发表时间：
2021-01-06
期刊：
影响因子：
4.8
通讯作者：
Xiao S
中科院分区：
文献类型：
--
作者：
Jin Z;Wei Q;Bi Y;Li Y;Huo N;Mou S;Wang W;Liu H;Yang Z;Chen H;Xiao S
纽卡斯尔病毒（NDV）的血凝素-神经氨酸酶（HN）蛋白是一种能诱导家禽产生保护性抗体的主要抗原。然而，其抗原表位尚未完全阐明。因此，确定HN的线性表位，特别是中和表位，将有助于揭示其抗原特性。在这项研究中，我们分析了B细胞免疫优势表位（IDEs）的HN蛋白从疫苗株LaSota使用pepscan技术与LaSota特异性的鸡超免疫抗血清。我们构建了IDEs-RFP质粒，制备了抗IDEs多肽小鼠血清，通过免疫学试验鉴定IDEs。最后，将不同稀释度的抗IDE抗血清用于BHK-21细胞进行中和试验。通过间接免疫荧光试验、斑点印迹和Western印迹法，用NDV和IDES特异性抗血清筛选并进一步验证了HN的5个IDES。所有5种IDE均显示出良好的免疫原性。IDE 5（328-342 aa）仅识别Ⅱ类NDV，与Ⅰ类NDV无反应。大多数IDE在不同菌株之间是高度保守的。体外中和试验表明，IDE 4（242-256 aa）和HN 341 -355（已报道的中和线性表位）的肽特异性鼠抗血清能部分中和LaSota无毒力株和强毒株，中和水平相近，提示IDE 4可能是一种潜在的中和线性表位。HN蛋白是NDV的主要保护性抗原，可在动物体内诱导中和抗体。我们确定了五个IDE的HN使用pepscan方法与NDV特异性鸡超免疫抗血清。5种IDE均能诱导小鼠产生特异性抗体。IDE 4（242-256 aa）被鉴定为新的潜在中和线性表位。这些结果将有助于阐明HN蛋白的抗原表位，为NDV疫苗的研制提供参考。
The hemagglutinin-neuraminidase (HN) protein of Newcastle disease virus (NDV) is a major antigen that can induce protective antibodies in poultry. However, its antigenic epitopes have not been fully elucidated. Therefore, defining the linear epitopes of HN, especially neutralizing epitopes, will be useful for revealing its antigenic characterization. In this study, we analyzed B-cell immunodominant epitopes (IDEs) of the HN protein from the vaccine strain LaSota using pepscan technology with LaSota-specific chicken hyperimmune antisera. We constructed IDEs-RFP plasmids and prepared anti-IDEs peptide mouse sera to identify IDEs through immunological tests. At last, the different diluted anti-IDE antisera were used in BHK-21 cells to perform the neutralization test. Five IDEs of the HN were screened and further verified by indirect immunofluorescence assays, dot blots and Western blots with NDV- and IDEs-specific antisera. All five IDEs showed good immunogenicity. IDE5 (328–342 aa) could recognize only class II NDV but did not react with the class I strain. Most of the IDEs are highly conserved among the different strains. A neutralization test in vitro showed that the peptide-specific mouse antisera of IDE4 (242–256 aa) and HN341-355, a reported neutralizing linear epitope, could partially neutralize avirulent LaSota as well as virulent strains at similar levels, suggesting that IDE4 might be a potential neutralizing linear epitope. The HN protein is a major protective antigen of NDV that can induce neutralizing antibodies in animals. We identified five IDEs of the HN using a pepscan approach with NDV-specific chicken hyperimmune antisera. The five IDEs could elicit specific antibodies in mice. IDE4 (242–256 aa) was identified as a novel potential neutralizing linear epitope. These results will help elucidate the antigenic epitopes of the HN and facilitate the development of NDV vaccines.
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