ABSTRACT

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.

中文翻译：

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EB病毒感染调节血脑屏障细胞及其与恶性疟原虫的共同感染诱导红细胞粘附

 抽象的

恶性疟原虫感染介导的 Epstein-Barr 病毒（EBV）再激活在疟疾流行国家已得到证实。我们假设，在疟疾发作期间，重新激活的 EBV 可以感染人脑微血管内皮细胞 (HBEC)。这可能会导致严重的脑部症状。我们在体外用 EBV 感染 HBEC。使用 qRT-PCR 分析随后的 EBV、炎症和内皮标记物的基因表达模式。此外，对维持血脑屏障 (BBB) 完整性的细胞进行伤口愈合测定，以研究 EBV 感染的 HBEC 分泌物的影响。进行红细胞粘附测定以评估 EBV 和恶性疟原虫单一感染和共同感染期间红细胞附着在 HBEC 上。我们的实验表明，HBEC 的 EBV 感染显着升高了多种炎症（TNFα、CCL2）和内皮（整合素 β3、PECAM、VEGFA、VWF、claudin-5、cx37）标记物。 EBV感染的HBEC分泌显着减少了HBEC、神经胶质细胞和神经元细胞的迁移。此外，与单一感染情况相比，EBV-恶性疟原虫共同感染显着（ P < 0.05）增强红细胞对 HBEC 的粘附。总之，HBEC 的 EBV 感染导致内皮激活并调节 BBB 微环境。 EBV-恶性疟原虫共同感染情况增加了红细胞对 EC 的粘附，这是脑型疟疾的一个标志。与疟疾一起，EBV 感染可能会加剧脑型疟疾病理学。