Department of Biosciences and Biomedical Engineering, Indian Institute of Technology Indore, Indore 453552, India.
Department of Microbiology and Immunology, National University of Singapore, Singapore 117545, Singapore.
Pathog Dis. 2021 Jan 9;79(1). doi: 10.1093/femspd/ftaa080.
Plasmodium falciparum infection-mediated Epstein-Barr virus (EBV) reactivation is well established in malaria-endemic countries. We hypothesize that, during malaria onset, the reactivated EBV can infect human brain microvascular endothelial cells (HBECs). This may cause severe cerebral manifestations. We infected HBECs with EBV in vitro. The subsequent gene expression pattern of EBV, inflammatory and endothelial markers was analysed using qRT-PCR. Further, a wound-healing assay for cells maintaining blood-brain barrier (BBB) integrity was performed to investigate the effect of EBV-infected HBECs secretions. The RBC adhesion assay was conducted to assess RBC attachment onto HBECs during EBV and P. falciparum mono- and co-infection. Our experiments revealed that EBV infection of HBECs significantly elevated several inflammatory (TNFα, CCL2) and endothelial (integrin β3, PECAM, VEGFA, VWF, claudin-5, cx37) markers. The EBV-infected HBECs secretion significantly reduced migration of HBECs, glial and neuronal cells. Additionally, EBV-P. falciparum co-infection significantly (P < 0.05) enhanced RBC adhesion to HBECs compared to mono-infection scenarios. Conclusively, the EBV infection of HBECs led to endothelial activation and modulated the BBB microenvironment. The EBV-P. falciparum co-infection scenario increased RBC adhesion on ECs which is a hallmark of cerebral malaria. Together with malaria, EBV infection can aid in exacerbation of cerebral malaria pathology.
疟原虫感染介导的 EBV（Epstein-Barr virus）再激活在疟疾流行国家已得到充分证实。我们假设，在疟疾发病期间，再激活的 EBV 可以感染人类脑微血管内皮细胞（HBECs），从而导致严重的脑表现。我们在体外感染 HBECs 中的 EBV。然后使用 qRT-PCR 分析 EBV、炎症和内皮标志物的后续基因表达模式。此外，还进行了维持血脑屏障（BBB）完整性的细胞划痕愈合试验，以研究 EBV 感染的 HBECs 分泌液的作用。进行 RBC 黏附试验，以评估 EBV 和疟原虫单感染及共感染期间 RBC 黏附到 HBECs 的情况。我们的实验表明，HBECs 中的 EBV 感染显著增加了几种炎症（TNFα、CCL2）和内皮（整合素β3、PECAM、VEGFA、VWF、claudin-5、cx37）标志物。EBV 感染的 HBECs 分泌显著降低了 HBECs、神经胶质细胞和神经元细胞的迁移。此外，与单感染情况相比，EBV-疟原虫共感染显著（P < 0.05）增加了 RBC 黏附到 HBECs 的情况。总之，EBV 感染 HBECs 导致内皮细胞激活并调节 BBB 微环境。EBV-疟原虫共感染情况增加了 RBC 黏附到 ECs 的情况，这是脑型疟疾的一个标志。与疟疾一起，EBV 感染可能有助于加重脑型疟疾的病理。
