Respiratory syncytial virus exhibits differential tropism for distinct human placental cell types with Hofbauer cells acting as a permissive reservoir for infection
Respiratory syncytial virus exhibits differential tropism for distinct human placental cell types with Hofbauer cells acting as a permissive reservoir for infection
复制标题
DOI：
10.1371/journal.pone.0225767
复制
发表时间：
2019-12-02
期刊：
影响因子：
3.7
通讯作者：
Piedimonte, Giovanni
中科院分区：
文献类型：
--
作者：
Bokun, Vladimir;Moore, John J.;Piedimonte, Giovanni
研究背景呼吸道合胞病毒（Respiratory syncytial virus，RSV）可引起短暂的病毒血症和肺外播散.最近，在胎儿脐带血中发现了这种病毒，这表明人类在子宫内获得的可能性。在这里，我们评估的perception和RSV复制的动力学在原代人胎盘细胞，检查他们的潜力，将这种感染转移到邻近的细胞，并测量引起的炎症反应的virus.Methods和findingsHuman胎盘绒毛组织收集后立即交付和胎盘细胞滋养层，成纤维细胞，巨噬细胞（Hofbauer）细胞的分离处理。用表达红色荧光蛋白（RFP）的重组RSV-A2毒株（rrRSV）感染分离的细胞，并通过荧光显微镜、蛋白质印迹和定量PCR（qPCR）进行分析。基于RFP表达，rrRSV表现出三种主要胎盘细胞类型的差异向性。胎盘成纤维细胞和Hofbauer细胞是允许的，并支持生产性rrRSV复制。虽然感染的细胞滋养层细胞表达病毒糖蛋白（G蛋白），但仅检测到有限的RSV复制。重要的是，qPCR和荧光聚焦单元测定显示，病毒子代在感染的Hofbauer细胞内滞留长达30天，没有释放到周围培养基中。然而，Hofbauer细胞将感染传递到覆盖的幼稚16 HBE细胞上，表明接触依赖性的反式感染。最后，通过多重细胞因子测定和常规ELISA在感染的Hofbauer细胞的上清液中测量到促炎细胞因子，特别是IL-6、TNF-α和IFN-γ的显著增加。并激发炎症反应。它还支持了这种呼吸道病毒可以垂直转移到胎儿并可能影响其发育和妊娠结果的假设。
BackgroundRespiratory syncytial virus (RSV) is capable of transient viremia and extrapulmonary dissemination. Recently, this virus has been identified in fetal cord blood, suggesting the possibility of in utero acquisition in humans. Here, we assess permissivity and kinetics of RSV replication in primary human placental cells, examine their potential to transfer this infection to neighboring cells, and measure the inflammatory response evoked by the virus.Methods and findingsHuman placental villus tissue was collected immediately upon delivery and processed for isolation of placental cytotrophoblast, fibroblast, and macrophage (Hofbauer) cells. Isolated cells were infected with a recombinant RSV-A2 strain (rrRSV) expressing red fluorescent protein (RFP) and analyzed by fluorescence microscopy, Western blot, and quantitative PCR (qPCR). Based on RFP expression, rrRSV exhibited differential tropism for the three major placental cell types. Placental fibroblasts and Hofbauer cells were permissive and supported productive rrRSV replication. While infected cytotrophoblast cells expressed viral glycoprotein (G protein), only limited RSV replication was detected. Importantly, qPCR and fluorescence-focused unit assay revealed that the viral progeny remains trapped within infected Hofbauer cells for up to 30 days, with no release into surrounding media. Yet, Hofbauer cells passed the infection onto overlaid naive 16HBE cells, suggesting contactdependent trans-infection. Lastly, a significant increase in proinflammatory cytokines, particularly IL-6, TNF-alpha, and IFN-gamma was measured in the supernatant of infected Hofbauer cells by multiplex cytokine assay and conventional ELISA.ConclusionsThis study demonstrates that RSV can replicate in human placenta, exhibits differential tropism for distinct placental cell types, can be stored and transferred to neighboring cells by Hofbauer cells, and elicits an inflammatory response. It also supports the hypothesis that this respiratory virus can be vertically transferred to the fetus and potentially affect its development and the outcome of pregnancies.