Introduction. Zika virus (ZIKV) emerged as a public health concern on the American continent during late 2015. As the number of infected grew so did the concerns about its capability to cause long-term damage especially with the appearance of the congenital Zika syndrome (CZS). Proteins from the TAM family of receptor tyrosine kinases (RTKs) were proposed as the cellular receptors, however, due to the ability of the virus to infect a variety of cell lines different strategies to elucidate the tropism of the virus should be investigated. Hypothesis. Pseudotyping is a powerful tool to interrogate the ability of the glycoprotein (GP) to permit entry of viruses. Aim. We aimed to establish a highly tractable pseudotype model using lenti- and retro-viral backbones to investigate the entry pathway of ZIKV. Methodology. We used different glycoprotein constructs and different lenti- or retro-viral backbones, in a matrix of ratios to investigate production of proteins and functional pseudotypes. Results. Varying the ratio of backbone and glycoprotein plasmids did not yield infectious pseudotypes. Moreover, the supplementation of the ZIKV protease or the substitution of the backbone had no positive impact on the infectivity. We showed production of the proteins in producer cells implying the lack of infectious pseudotypes is due to a lack of successful glycoprotein incorporation, rather than lack of protein production. Conclusion. In line with other reports, we were unable to successfully produce infectious pseudotypes using the variety of methods described. Other strategies may be more suitable in the development of an efficient pseudotype model for ZIKV and other flaviviruses.

中文翻译：

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开发寨卡糖蛋白假型分析面临的挑战


介绍。 2015 年底，寨卡病毒 (ZIKV) 成为美洲大陆的一个公共卫生问题。随着感染人数的增加，人们对其造成长期损害的能力的担忧也随之增加，尤其是先天性寨卡综合症 (CZS) 的出现。来自受体酪氨酸激酶 (RTK) 的 TAM 家族的蛋白质被提议作为细胞受体，然而，由于病毒能够感染多种细胞系，因此应该研究不同的策略来阐明病毒的向性。假设。假型分析是检测糖蛋白 (GP) 允许病毒进入的能力的强大工具。目的。我们的目标是使用慢病毒和逆转录病毒骨架建立高度易处理的假型模型来研究 ZIKV 的进入途径。方法论。我们在比例矩阵中使用不同的糖蛋白构建体和不同的慢病毒或逆转录病毒主链来研究蛋白质和功能假型的产生。结果。改变骨架和糖蛋白质粒的比例并没有产生感染性假型。此外，补充 ZIKV 蛋白酶或替换主链对感染性没有积极影响。我们展示了生产细胞中蛋白质的产生，这意味着缺乏感染性假型是由于缺乏成功的糖蛋白掺入，而不是缺乏蛋白质产生。结论。与其他报告一致，我们无法使用所描述的各种方法成功地产生感染性假型。 其他策略可能更适合开发 ZIKV 和其他黄病毒的有效假型模型。