Efficient generation of vesicular stomatitis virus (VSV)-pseudotypes bearing morbilliviral glycoproteins and their use in quantifying virus neutralising antibodies.
Efficient generation of vesicular stomatitis virus (VSV)-pseudotypes bearing morbilliviral glycoproteins and their use in quantifying virus neutralising antibodies.
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DOI：
10.1016/j.vaccine.2015.12.006
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发表时间：
2016-02-03
期刊：
影响因子：
5.5
通讯作者：
Willett BJ
中科院分区：
文献类型：
--
作者：
Logan N;McMonagle E;Drew AA;Takahashi E;McDonald M;Baron MD;Gilbert M;Cleaveland S;Haydon DT;Hosie MJ;Willett BJ
麻疹病毒中和抗体传统上使用空斑减少中和试验（PRNT）或活病毒微中和试验（micro-NT）来测量。虽然这两种测试形式都可以对体液反应的强度和特异性进行可靠的评估，但它们受到可研究的病毒株数量有限的限制，并且常常给操作者带来重大的生物安全问题。在这项研究中，我们描述了基于复制缺陷型水泡性口炎病毒（VSVΔG）的假型系统的适应，用于测量麻疹病毒中和抗体。通过在带有荧光素酶标记基因的 VSVΔG 假型上表达犬瘟热病毒 (CDV) 的血凝素 (H) 和融合 (F) 蛋白，可以快速、高灵敏度地测量中和抗体滴度。此外，通过交换糖蛋白表达构建体，可以测量针对不同病毒株或物种的反应。使用这种技术，我们证明了 CDV 和小反刍兽疫病毒 (PPRV) 之间的交叉中和作用。作为该技术价值的一个例子，我们证明了英国狗在 CDV 疫苗接种诱导的免疫广度上存在差异；在一些狗中，中和反应是 CDV 特异性的，而在另一些狗中，中和反应延伸到反刍动物麻疹病毒 PPRV。该技术将有助于对麻疹病毒进行交叉中和的全面比较。
Morbillivirus neutralising antibodies are traditionally measured using either plaque reduction neutralisation tests (PRNTs) or live virus microneutralisation tests (micro-NTs). While both test formats provide a reliable assessment of the strength and specificity of the humoral response, they are restricted by the limited number of viral strains that can be studied and often present significant biological safety concerns to the operator. In this study, we describe the adaptation of a replication-defective vesicular stomatitis virus (VSVΔG) based pseudotyping system for the measurement of morbillivirus neutralising antibodies. By expressing the haemagglutinin (H) and fusion (F) proteins of canine distemper virus (CDV) on VSVΔG pseudotypes bearing a luciferase marker gene, neutralising antibody titres could be measured rapidly and with high sensitivity. Further, by exchanging the glycoprotein expression construct, responses against distinct viral strains or species may be measured. Using this technique, we demonstrate cross neutralisation between CDV and peste des petits ruminants virus (PPRV). As an example of the value of the technique, we demonstrate that UK dogs vary in the breadth of immunity induced by CDV vaccination; in some dogs the neutralising response is CDV-specific while, in others, the neutralising response extends to the ruminant morbillivirus PPRV. This technique will facilitate a comprehensive comparison of cross-neutralisation to be conducted across the morbilliviruses.