Frumence Etienne, Viranaicken Wildriss, Gadea Gilles, Desprès Philippe
Université de La Réunion, INSERM U1187, CNRS UMR 9192, IRD UMR 249, Unité Mixte Processus Infectieux en Milieu Insulaire Tropical, Plateforme Technologique CYROI, 97491 Sainte-Clotilde, La Réunion, France.
Vaccines (Basel). 2019 Jul 16;7(3):66. doi: 10.3390/vaccines7030066.
Zika virus (ZIKV) is an emerging arthropod-borne virus of major public health concern. ZIKV infection is responsible for congenital Zika disease and other neurological defects. Antibody-mediated virus neutralization is an essential component of protective antiviral immunity against ZIKV. In the present study, we assessed whether our GFP reporter ZIKV derived from African viral strain MR766 could be useful for the development of a flow cytometry neutralization test (FNT), as an alternative to the conventional plaque-reduction neutralization test (PRNT). To improve the efficacy of GFP-expressing MR766, we selected virus variant MR766 showing a high level of GFP signal in infected cells. A MR766-based FNT was assayed with immune sera from adult mice that received ZIKBeHMR-2. The chimeric ZIKV clone ZIKBeHMR-2 comprises the structural protein region of epidemic strain BeH819015 into MR766 backbone. We reported that adult mice inoculated with ZIKBeHMR-2 developed high levels of neutralizing anti-ZIKV antibodies. Comparative analysis between MR766-based FNT and conventional PRNT was performed using mouse anti-ZIKBeHMR-2 immune sera. Indistinguishable neutralization patterns were observed when compared with PRNT and FNT. We consider that the newly developed MR766-based FNT is a valid format for measuring ZIKV-neutralizing antibodies in serum specimens.
寨卡病毒（ZIKV）是一种新出现的节肢动物传播病毒，引起了重大的公共卫生关注。寨卡病毒感染会导致先天性寨卡疾病和其他神经缺陷。抗体介导的病毒中和是针对寨卡病毒的保护性抗病毒免疫的重要组成部分。在本研究中，我们评估了源自非洲病毒株MR766的绿色荧光蛋白（GFP）报告基因寨卡病毒是否可用于开发流式细胞术中和试验（FNT），作为传统蚀斑减少中和试验（PRNT）的替代方法。为了提高表达GFP的MR766的效力，我们选择了在感染细胞中显示高水平GFP信号的病毒变体MR766。基于MR766的FNT用接受ZIKBeHMR - 2的成年小鼠的免疫血清进行检测。嵌合寨卡病毒克隆ZIKBeHMR - 2将流行株BeH819015的结构蛋白区域包含到MR766骨架中。我们报道，接种ZIKBeHMR - 2的成年小鼠产生了高水平的中和抗寨卡病毒抗体。使用小鼠抗ZIKBeHMR - 2免疫血清对基于MR766的FNT和传统PRNT进行了比较分析。与PRNT和FNT相比，观察到了难以区分的中和模式。我们认为新开发的基于MR766的FNT是一种用于测量血清标本中寨卡病毒中和抗体的有效方法。
