Hantaan virus can infect human keratinocytes and activate an interferon response through the nuclear translocation of IRF-3

Ye, Wei; Xu, Yongni; Wang, Yuan; Dong, Yangchao; Xi, Qianqian; Cao, Mengyuan; Lan, Yu; Zhang, Liang; Cheng, Long; Wu, Xingan; Zhang, Xu; Lei, Yingfeng; Zhang, Fanglin

doi:10.1016/j.meegid.2014.11.009

Cited by 16 publications

(11 citation statements)

References 78 publications

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“…HTNV titers are commonly measured by TCID50 with Reed and Muench's formula through ELISA (Ye et al, 2015b). The positive or negative infection wells in the ELISA are determined by the P (positive)/N (negative) value of HTNV NP, which indicates that the negative wells have P/N values ≤ 2.1 and the positive wells have P/N values ≥ 2.1.…”

Section: Resultsmentioning

confidence: 99%

“…The improved plaque formation test is dependent on the low pH-induced cytopathic effects of hantavirus but is time-consuming and has low reproducibility. The most widely adopted approach to test hantavirus titers (especially for HTNV) is the TCID50 (50% tissue culture infective dose) calculation using ELISA as previously reported by our group (Xu et al, 2002; Cheng et al, 2014; Jiang et al, 2015; Ye et al, 2015a,b; Ying et al, 2016); however, virus propagation in Vero E6 cells takes at least 10 days. All the reported detective measurements have insurmountably objective drawbacks, such as high demanding experimental conditions for qRT-PCR and expensive apparatus and labware for FCM, which limits their applicability (Wan et al, 2010).…”

Section: Introductionmentioning

confidence: 99%

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In-Cell Western Assays to Evaluate Hantaan Virus Replication as a Novel Approach to Screen Antiviral Molecules and Detect Neutralizing Antibody Titers

Chen

et al. 2017

Front. Cell. Infect. Microbiol.

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Hantaviruses encompass rodent-borne zoonotic pathogens that cause severe hemorrhagic fever disease with high mortality rates in humans. Detection of infectious virus titer lays a solid foundation for virology and immunology researches. Canonical methods to assess viral titers rely on visible cytopathic effects (CPE), but Hantaan virus (HTNV, the prototype hantavirus) maintains a relatively sluggish life cycle and does not produce CPE in cell culture. Here, an in-cell Western (ICW) assay was utilized to rapidly measure the expression of viral proteins in infected cells and to establish a novel approach to detect viral titers. Compared with classical approaches, the ICW assay is accurate and time- and cost-effective. Furthermore, the ICW assay provided a high-throughput platform to screen and identify antiviral molecules. Potential antiviral roles of several DExD/H box helicase family members were investigated using the ICW assay, and the results indicated that DDX21 and DDX60 reinforced IFN responses and exerted anti-hantaviral effects, whereas DDX50 probably promoted HTNV replication. Additionally, the ICW assay was also applied to assess NAb titers in patients and vaccine recipients. Patients with prompt production of NAbs tended to have favorable disease outcomes. Modest NAb titers were found in vaccinees, indicating that current vaccines still require improvements as they cannot prime host humoral immunity with high efficiency. Taken together, our results indicate that the use of the ICW assay to evaluate non-CPE Hantaan virus titer demonstrates a significant improvement over current infectivity approaches and a novel technique to screen antiviral molecules and detect NAb efficacies.

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Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

In-Cell Western Assays to Evaluate Hantaan Virus Replication as a Novel Approach to Screen Antiviral Molecules and Detect Neutralizing Antibody Titers

Chen

et al. 2017

Front. Cell. Infect. Microbiol.

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“…The mRNA expression level of each target gene was normalized to the respective β-actin and analyzed. The qRT-PCR primer sequences for NEAT1, NEAT1-2, IFN-β, HTNV S segment, RIG-I, DDX60, β-actin, and GAPDH were obtained from previous reports (24, 45). The methods used to quantify HTNV RNA load have been described by our group previously (46).…”

Section: Methodsmentioning

confidence: 99%

The Long Noncoding RNA NEAT1 Exerts Antihantaviral Effects by Acting as Positive Feedback for RIG-I Signaling

Han

et al. 2017

J Virol

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155

167

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Hantavirus infection, which causes zoonotic diseases with a high mortality rate in humans, has long been a global public health concern. Over the past decades, accumulating evidence suggests that long noncoding RNAs (lncRNAs) play key regulatory roles in innate immunity. However, the involvement of host lncRNAs in hantaviral control remains uncharacterized. In this study, we identified the lncRNA NEAT1 as a vital antiviral modulator. NEAT1 was dramatically upregulated after Hantaan virus (HTNV) infection, whereas its downregulation in vitro or in vivo delayed host innate immune responses and aggravated HTNV replication. Ectopic expression of NEAT1 enhanced beta interferon (IFN-β) production and suppressed HTNV infection. Further investigation suggested that NEAT1 served as positive feedback for RIG-I signaling. HTNV infection activated NEAT1 transcription through the RIG-I–IRF7 pathway, whereas NEAT1 removed the transcriptional inhibitory effects of the splicing factor proline- and glutamine-rich protein (SFPQ) by relocating SFPQ to paraspeckles, thus promoting the expression of RIG-I and DDX60. RIG-I and DDX60 had synergic effects on IFN production. Taken together, our findings demonstrate that NEAT1 modulates the innate immune response against HTNV infection, providing another layer of information about the role of lncRNAs in controlling viral infections.IMPORTANCE Hantaviruses have attracted worldwide attention as archetypal emerging pathogens. Recently, increasing evidence has highlighted long noncoding RNAs (lncRNAs) as key regulators of innate immunity; however, their roles in hantavirus infection remain unknown. In the present work, a new unexplored function of lncRNA NEAT1 in controlling HTNV replication was found. NEAT1 promoted interferon (IFN) responses by acting as positive feedback for RIG-I signaling. This lncRNA was induced by HTNV through the RIG-I–IRF7 pathway in a time- and dose-dependent manner and promoted HTNV-induced IFN production by facilitating RIG-I and DDX60 expression. Intriguingly, NEAT1 relocated SFPQ and formed paraspeckles after HTNV infection, which might reverse inhibitive effects of SFPQ on the transcription of RIG-I and DDX60. To the best of our knowledge, this is the first study to address the regulatory role of the lncRNA NEAT1 in host innate immunity after HTNV infection. In summary, our findings provide additional insights regarding the role of lncRNAs in controlling viral infections.

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“…Despite a low replication rate and release of infectious virus due to inhibition by IFN-α, monocytes/macrophages from peripheral blood are susceptible to PUUV infection [61,62]. Recently, keratinocytes have also been shown to be permissive to hantavirus infection [63], which is of interest considering that hantaviruses can be transmitted by bites between small mammals.…”

Section: Hantavirus Propagation In Different Cell Typesmentioning

confidence: 99%

“…Proteomics has revealed a high degree of CXCL10 activation in HuH7 cells infected with PUUV, as compared to non-infected control cells or cells infected with nonpathogenic TULV or PHV (our unpublished data). CCL5 and CXCL10 were also up-regulated in keratinocytes derived cells that are susceptible to HTNV [63]. …”

Section: Activation Of Host Cell Factors By Hantavirusesmentioning

confidence: 99%

What Do We Know about How Hantaviruses Interact with Their Different Hosts?

Ermonval

Baychelier

Tordo

2016

Viruses

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Hantaviruses, like other members of the Bunyaviridae family, are emerging viruses that are able to cause hemorrhagic fevers. Occasional transmission to humans is due to inhalation of contaminated aerosolized excreta from infected rodents. Hantaviruses are asymptomatic in their rodent or insectivore natural hosts with which they have co-evolved for millions of years. In contrast, hantaviruses cause different pathologies in humans with varying mortality rates, depending on the hantavirus species and its geographic origin. Cases of hemorrhagic fever with renal syndrome (HFRS) have been reported in Europe and Asia, while hantavirus cardiopulmonary syndromes (HCPS) are observed in the Americas. In some cases, diseases caused by Old World hantaviruses exhibit HCPS-like symptoms. Although the etiologic agents of HFRS were identified in the early 1980s, the way hantaviruses interact with their different hosts still remains elusive. What are the entry receptors? How do hantaviruses propagate in the organism and how do they cope with the immune system? This review summarizes recent data documenting interactions established by pathogenic and nonpathogenic hantaviruses with their natural or human hosts that could highlight their different outcomes.

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Hantaan virus can infect human keratinocytes and activate an interferon response through the nuclear translocation of IRF-3

Cited by 16 publications

References 78 publications

In-Cell Western Assays to Evaluate Hantaan Virus Replication as a Novel Approach to Screen Antiviral Molecules and Detect Neutralizing Antibody Titers

In-Cell Western Assays to Evaluate Hantaan Virus Replication as a Novel Approach to Screen Antiviral Molecules and Detect Neutralizing Antibody Titers

The Long Noncoding RNA NEAT1 Exerts Antihantaviral Effects by Acting as Positive Feedback for RIG-I Signaling

What Do We Know about How Hantaviruses Interact with Their Different Hosts?

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