Integrated Research Facility, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Ft. Detrick, Frederick, Maryland, United States of America.
Department of Microbiology and Immunology, The University of Maryland School of Medicine, Baltimore, MD, United States of America.
PLoS One. 2018 Mar 22;13(3):e0194868. doi: 10.1371/journal.pone.0194868. eCollection 2018.
Middle East respiratory syndrome coronavirus (MERS-CoV) presents an emerging threat to public health worldwide by causing severe respiratory disease in humans with high virulence and case fatality rate (about 35%) since 2012. Little is known about the pathogenesis and innate antiviral response in primary human monocyte-derived macrophages (MDMs) and dendritic cells (MDDCs) upon MERS-CoV infection. In this study, we assessed MERS-CoV replication as well as induction of inflammatory cytokines and chemokines in MDMs and immature and mature MDDCs. Immature MDDCs and MDMs were permissive for MERS-CoV infection, while mature MDDCs were not, with stimulation of proinflammatory cytokine and chemokine upregulation in MDMs, but not in MDDCs. To further evaluate the antiviral activity of well-defined drugs in primary antigen presenting cells (APCs), three compounds (chloroquine, chlorpromazine and toremifine), each with broad-spectrum antiviral activity in immortalized cell lines, were evaluated in MDMs and MDDCs to determine their antiviral effect on MERS-CoV infection. While chloroquine was not active in these primary cells, chlorpromazine showed strong anti-MERS-CoV activity, but it was associated with high cytotoxicity narrowing the potential window for drug utilization. Unlike in established cells, toremifene had marginal activity when tested in antigen presenting cells, with high apparent cytotoxicity, also limiting its potential as a therapeutic option. These results demonstrate the value of testing drugs in primary cells, in addition to established cell lines, before initiating preclinical or clinical studies for MERS treatment and the importance of carefully assessing cytotoxicity in drug screen assays. Furthermore, these studies also highlight the role of APCs in stimulating a robust protective immune response to MERS-CoV infection.
中东呼吸综合征冠状病毒(MERS-CoV)自 2012 年以来在人类中引起严重的呼吸道疾病,具有高毒力和高病死率(约 35%),对全球公共卫生构成了新的威胁。目前人们对原发性人单核细胞衍生的巨噬细胞(MDMs)和树突状细胞(MDDCs)中 MERS-CoV 感染的发病机制和先天抗病毒反应知之甚少。在本研究中,我们评估了 MERS-CoV 在 MDMs 和未成熟及成熟 MDDCs 中的复制以及诱导的炎症细胞因子和趋化因子。未成熟的 MDDCs 和 MDMs 允许 MERS-CoV 感染,而成熟的 MDDCs 则不允许,这会刺激 MDMs 中促炎细胞因子和趋化因子的上调,但不会在 MDDCs 中上调。为了进一步评估在原代抗原呈递细胞(APCs)中明确药物的抗病毒活性,评估了三种化合物(氯喹、氯丙嗪和托瑞米芬),它们在永生化细胞系中均具有广谱抗病毒活性,以确定它们对 MERS-CoV 感染的抗病毒作用。虽然氯喹在这些原代细胞中无效,但氯丙嗪显示出很强的抗 MERS-CoV 活性,但它与高细胞毒性相关,缩小了药物利用的潜在窗口。与在已建立的细胞中不同,在抗原呈递细胞中测试时,托瑞米芬的活性很微弱,同时具有高细胞毒性,这也限制了它作为治疗选择的潜力。这些结果表明,在开始进行 MERS 治疗的临床前或临床研究之前,除了建立细胞系外,在原代细胞中测试药物非常有价值,并且在药物筛选试验中仔细评估细胞毒性非常重要。此外,这些研究还强调了 APCs 在刺激针对 MERS-CoV 感染的强大保护性免疫反应中的作用。