吡丙醚对病毒行为的影响：对杀虫剂诱导毒力和传播机制的影响。
Impact of pyriproxyfen on virus behavior: implications for pesticide-induced virulence and mechanism of transmission.
机构信息
Dr. Kiran C. Patel College of Osteopathic Medicine, Nova Southeastern University, 3400 Gulf to Bay Blvd, Clearwater, FL, 33759, USA.
Department of Biological Sciences, Halmos College of Arts and Sciences, Nova Southeastern University, Fort Lauderdale, FL, 33314, USA.
出版信息
Virol J. 2020 Jul 6;17(1):93. doi: 10.1186/s12985-020-01378-y.
BACKGROUND
More than 3 years since the last Zika virus (ZIKV) outbreak in Brazil, researchers are still deciphering the molecular mechanisms of neurovirulence and vertical transmission, as well as the best way to control spread of ZIKV, a flavivirus. The use of pesticides was the main strategy of mosquito control during the last ZIKV outbreak.
METHODS
We used vesicular stomatitis virus (VSV) tagged with green fluorescent protein (GFP) as our prototypical virus to study the impact of insecticide pyriproxyfen (PPF). VZV-GFP infected and uninfected Jurkat, HeLa and trophoblast cells were treated with PPF and compared to untreated cells (control). Cell viability was determined by the MTT assay. Cell morphology, presence of extracellular vesicles (EVs), virus infection/GFP expression as well as active mitochondrial levels/localization were examined by confocal microscopy.
RESULTS
PPF, which was used to control mosquito populations in Brazil prior to the ZIKV outbreak, enhances VSV replication and has cell membrane-altering properties in the presence of virus. PPF causes enhanced viral replication and formation of large EVs, loaded with virus as well as mitochondria. Treatment of trophoblasts or HeLa cells with increasing concentrations of PPF does not alter cell viability, however, it proportionately increases Jurkat cell viability. Increasing concentrations of PPF followed by VSV infection does not interfere with HeLa cell viability. Both Jurkats and trophoblasts show proportionately increased cell death with increased concentrations of PPF in the presence of virus.
CONCLUSIONS
We hypothesize that PPF disrupts the lipid microenvironment of mammalian cells, thereby interfering with pathways of viral replication. PPF lowers viability of trophoblasts and Jurkats in the presence of VSV, implying that the combination renders immune system impairment in infected individuals as well as enhanced vulnerability of fetuses towards viral vertical transmission. We hypothesize that similar viruses such as ZIKV may be vertically transmitted via EV-to-cell contact when exposed to PPF, thereby bypassing immune detection. The impact of pesticides on viral replication must be fully investigated before large scale use in future outbreaks of mosquito borne viruses.
背景
自巴西上一次寨卡病毒（ZIKV）爆发以来已有 3 年多，研究人员仍在破译神经毒力和垂直传播的分子机制，以及控制寨卡病毒（一种黄病毒）传播的最佳方法。在上一次寨卡病毒爆发期间，使用杀虫剂是控制蚊子的主要策略。
方法
我们使用带有绿色荧光蛋白（GFP）的水疱性口炎病毒（VSV）作为我们的原型病毒来研究杀虫剂吡丙醚（PPF）的影响。用 PPF 处理感染和未感染 VZV-GFP 的 Jurkat、HeLa 和滋养层细胞，并与未处理的细胞（对照）进行比较。通过 MTT 测定法确定细胞活力。通过共聚焦显微镜检查细胞形态、细胞外囊泡（EVs）的存在、病毒感染/GFP 表达以及活性线粒体水平/定位来评估细胞活力。
结果
在寨卡病毒爆发之前，用于控制巴西蚊子种群的杀虫剂吡丙醚在存在病毒的情况下增强了 VSV 的复制并具有改变细胞膜的特性。PPF 导致病毒复制增强和形成大的 EV，这些 EV 装载有病毒和线粒体。用逐渐增加浓度的 PPF 处理滋养层细胞或 HeLa 细胞不会改变细胞活力，但会相应增加 Jurkat 细胞活力。用逐渐增加浓度的 PPF 处理 Jurkats 和滋养层细胞，然后用 VSV 感染，不会干扰 HeLa 细胞活力。在病毒存在的情况下，用逐渐增加浓度的 PPF 处理 Jurkats 和滋养层细胞均会导致细胞死亡比例增加。
结论
我们假设 PPF 破坏了哺乳动物细胞的脂质微环境，从而干扰了病毒复制途径。在 VSV 存在的情况下，PPF 降低了滋养层细胞和 Jurkat 细胞的活力，这意味着在感染个体中，这种组合会导致免疫系统受损，并且胎儿对病毒垂直传播的脆弱性增加。我们假设，当接触 PPF 时，类似寨卡病毒的病毒可能通过 EV 与细胞的接触进行垂直传播，从而逃避免疫检测。在未来的蚊子传播病毒爆发中大规模使用杀虫剂之前，必须充分研究其对病毒复制的影响。
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