Valdés-López Juan Felipe, Tamayo-Molina Yordi Sebastián, Fernandez Geysson J, Hernández-Sarmiento Lady Johana, Velilla Paula A, Urcuqui-Inchima Silvio
Grupo Inmunovirología, Facultad de Medicina, UdeA, Medellín, Colombia.
PLoS Negl Trop Dis. 2025 Sep 11;19(9):e0013366. doi: 10.1371/journal.pntd.0013366. eCollection 2025 Sep.
A Cytokine storm is critical in severe dengue, significantly contributing to disrupted endothelial integrity, plasma leakage, and haemorrhage manifestations in affected patients. Various reports have demonstrated that mononuclear phagocytes, including monocytes, dendritic cells, and macrophages, are target cells of DENV infection. They contribute to viral spread into tissues and promote robust inflammatory responses and immunopathology. However, it remains unclear whether the early events of DENV infection play a role in triggering cytokine storms in infected mononuclear phagocytes. To address this knowledge gap, we conducted a comprehensive analysis of the transcriptomic profile of in vitro DENV-2-infected human monocyte-derived macrophages (MDMs) based on the kinetics of viral replication through a standard growth curve. To verify the accuracy of our approach, we used RT-qPCR, ELISA, and transcriptomic data from in vitro DENV-2-infected monocyte-derived dendritic cells (MDDCs) and monocytes obtained from acute dengue patients. RNA-Seq analysis revealed dynamic changes in the transcriptional profile of DENV-2-infected MDMs throughout the viral growth curve. Two waves of differentially expressed genes were observed: the first occurred during the eclipse period of viral replication (3 to 5.5 h.p.i) and was associated with the induction of NF-kB-dependent pro-inflammatory factors, while the second wave at 24 h.p.i coincided with peaks of DENV-2 replication and induction of both NF-kB- and STAT1-dependent pro-inflammatory responses. Additionally, DENV-2 infection promoted the dynamic activation of Toll-like receptors, RIG-like receptors, inflammasomes, and inflammatory pathways, triggering innate pro-inflammatory and antiviral responses. A robust multi-type IFN-dependent antiviral response was also observed at the late stage of infection. A similar transcriptomic profile was found in DENV-2-infected MDDCs and monocyte subsets from acute dengue patients, further confirming the reliability of our in vitro model of DENV-infected MDMs. Together, results suggest that recognizing viral PAMPs during the eclipse period of DENV-2 infection promotes a robust NF-kB-dependent pro-inflammatory response in MDMs. In addition, at later stages of infection, recognizing structural DENV-PAMPs and/or viral replication intermediates induces both NF-kB- and STAT1-dependent pro-inflammatory responses, leading to a cytokine storm. These findings highlight the critical role of monocytes, macrophages, and dendritic cells in detecting DENV infection and triggering a cytokine storm in vitro and in vivo. This suggests that these cell populations could be potential targets for future immunotherapies to modulate the inflammatory response to DENV infection.
细胞因子风暴在严重登革热中至关重要，在受影响患者中显著导致内皮完整性破坏、血浆渗漏和出血表现。各种报告表明，包括单核细胞、树突状细胞和巨噬细胞在内的单核吞噬细胞是登革病毒感染的靶细胞。它们有助于病毒扩散到组织中，并促进强烈的炎症反应和免疫病理学。然而，登革病毒感染的早期事件是否在触发受感染单核吞噬细胞中的细胞因子风暴中起作用仍不清楚。为了填补这一知识空白，我们基于病毒复制动力学通过标准生长曲线对体外登革病毒2型感染的人单核细胞衍生巨噬细胞（MDM）的转录组谱进行了全面分析。为了验证我们方法的准确性，我们使用了来自体外登革病毒2型感染的单核细胞衍生树突状细胞（MDDC）和急性登革热患者单核细胞的RT-qPCR、ELISA和转录组数据。RNA测序分析揭示了登革病毒2型感染的MDM在整个病毒生长曲线中的转录谱动态变化。观察到两波差异表达基因：第一波发生在病毒复制的隐蔽期（感染后3至5.5小时），与NF-κB依赖性促炎因子的诱导有关，而第二波在感染后24小时与登革病毒2型复制峰值以及NF-κB和STAT1依赖性促炎反应的诱导同时出现。此外，登革病毒2型感染促进了Toll样受体、RIG样受体、炎性小体和炎症途径的动态激活，触发先天性促炎和抗病毒反应。在感染后期还观察到强大的多类型IFN依赖性抗病毒反应。在登革病毒2型感染的MDDC和急性登革热患者的单核细胞亚群中发现了类似的转录组谱，进一步证实了我们的登革病毒感染MDM体外模型的可靠性。总之，结果表明在登革病毒2型感染的隐蔽期识别病毒PAMP可促进MDM中强大的NF-κB依赖性促炎反应。此外，在感染后期，识别结构性登革病毒PAMP和/或病毒复制中间体可诱导NF-κB和STAT1依赖性促炎反应，导致细胞因子风暴。这些发现突出了单核细胞、巨噬细胞和树突状细胞在检测登革病毒感染以及在体外和体内触发细胞因子风暴中的关键作用。这表明这些细胞群体可能是未来免疫疗法调节对登革病毒感染的炎症反应的潜在靶点。
