Identification of Host Factors Differentially Induced by Clinically Diverse Strains of Tick-Borne Encephalitis Virus

Goonawardane, Niluka; Upstone, Laura; Harris, Mark; Jones, Ian M.

doi:10.1128/jvi.00818-22

Cited by 6 publications

(14 citation statements)

References 73 publications

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“…coli DH5α (Promega) and purified with commercially available plasmid DNA isolation kits as per the manufacturer’s protocols (Qiagen). Point mutations and nucleotide substitutions were introduced into the infectious clone (IC) or TBEV replicon [ 17 ], using Q5 site-directed mutagenesis kits (NEB). All primers are shown in Table 1 .…”

Section: Methodsmentioning

confidence: 99%

“…The construction of the TBEV replicons has been described elsewhere [ 17 ]. Briefly, most structural genes (nts 183–2386) were removed to make them non-infectious.…”

Section: Methodsmentioning

confidence: 99%

“…The 5’UTR sequence and Stem-loops (SLs) 3 and 4 were retained in the C-terminus of the C-protein due to their requirement during the formation of secondary RNA structures essential for genome cyclization. Luciferase was cloned downstream of the 5’-end of the C-protein to provide a marker of both replication and translation as previously described [ 17 ]. Dinucleotides were modified (no CpGs and low UpA frequency[cu]) to enhance replication capacity [ 17 , 27 ].…”

Section: Methodsmentioning

confidence: 99%

“…Luciferase was cloned downstream of the 5’-end of the C-protein to provide a marker of both replication and translation as previously described [ 17 ]. Dinucleotides were modified (no CpGs and low UpA frequency[cu]) to enhance replication capacity [ 17 , 27 ]. Firefly luciferase contained no CpGs and low UpAs (termed luc-cu).…”

Section: Methodsmentioning

confidence: 99%

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Functional characterization of 5′ untranslated region (UTR) secondary RNA structures in the replication of tick-borne encephalitis virus in mammalian cells

et al. 2023

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Tick-borne Encephalitis Virus (TBEV) is an emerging flavivirus that causes neurological disorders including viral encephalitis of varying severity. Whilst secondary RNA structures within the 5′ untranslated regions (UTRs) of many flaviviruses determine both virus replication and pathogenic outcomes in humans, these elements have not been systematically investigated for TBEV. In this study, we investigated the role of predicted RNA secondary elements of the first 107 nucleotides (nts) of the viral genome forming the stem-loop A (SLA). Experiments were performed in replicons and infectious TBEV system. This region comprises three distinct structures: 5’ stem 0 (S0), stem-loop 1 (SL1) and stem-loop 2 (SL2). S0 was found to be essential for virus infection as mutations in the lower stem of this region significantly reduced virus replication. Point mutations in SL1 that preserved the Y-shape confirmation delayed viral RNA replication but did not abolish virus infectivity. Deletion of SL2 did not abolish infectivity but had a negligible effect on virus propagation. No correlation was observed between in vitro translation efficiency and virus infectivity, suggesting that the 5’UTR functions independently to virus translation. Together, these findings reveal distinct RNA elements within the 5′UTR that are essential for the stability and replication of viral RNA. We further identify changes in RNA folding that lead to altered TBEV infectivity and pathogenesis.

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Section: Methodsmentioning

confidence: 99%

“…The construction of the TBEV replicons has been described elsewhere [ 17 ]. Briefly, most structural genes (nts 183–2386) were removed to make them non-infectious.…”

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

See 2 more Smart Citations

Functional characterization of 5′ untranslated region (UTR) secondary RNA structures in the replication of tick-borne encephalitis virus in mammalian cells

et al. 2023

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“…It has been shown in many studies that the determinants of viral neuropathogenesis were located in the E protein [ 12 , 13 ]. In addition, mutations in the NS5 protein have been shown to contribute to the attenuation of virulence and host–cell interactions [ 13 , 19 , 20 ]. Amino acid substitutions in NS3 are known to affect the budding of viral particles, thus leading to diminished pathogenicity [ 11 , 21 ].…”

Section: Introductionmentioning

confidence: 99%

Activation of Early Proinflammatory Responses by TBEV NS1 Varies between the Strains of Various Subtypes

Starodubova

Tuchynskaya

Kuzmenko

et al. 2023

IJMS

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Tick-borne encephalitis (TBE) is an emerging zoonosis that may cause long-term neurological sequelae or even death. Thus, there is a growing interest in understanding the factors of TBE pathogenesis. Viral genetic determinants may greatly affect the severity and consequences of TBE. In this study, nonstructural protein 1 (NS1) of the tick-borne encephalitis virus (TBEV) was tested as such a determinant. NS1s of three strains with similar neuroinvasiveness belonging to the European, Siberian and Far-Eastern subtypes of TBEV were studied. Transfection of mouse cells with plasmids encoding NS1 of the three TBEV subtypes led to different levels of NS1 protein accumulation in and secretion from the cells. NS1s of TBEV were able to trigger cytokine production either in isolated mouse splenocytes or in mice after delivery of NS1 encoding plasmids. The profile and dynamics of TNF-α, IL-6, IL-10 and IFN-γ differed between the strains. These results demonstrated the involvement of TBEV NS1 in triggering an immune response and indicated the diversity of NS1 as one of the genetic factors of TBEV pathogenicity.

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Autoantibodies neutralizing type I IFNs underlie severe tick-borne encephalitis in ∼10% of patients

Gervais,

Marchal,

Fortova

et al. 2024

Journal of Experimental Medicine

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Tick-borne encephalitis (TBE) virus (TBEV) is transmitted to humans via tick bites. Infection is benign in >90% of the cases but can cause mild (<5%), moderate (<4%), or severe (<1%) encephalitis. We show here that ∼10% of patients hospitalized for severe TBE in cohorts from Austria, Czech Republic, and France carry auto-Abs neutralizing IFN-α2, -β, and/or -ω at the onset of disease, contrasting with only ∼1% of patients with moderate and mild TBE. These auto-Abs were found in two of eight patients who died and none of 13 with silent infection. The odds ratios (OR) for severe TBE in individuals with these auto-Abs relative to those without them in the general population were 4.9 (95% CI: 1.5–15.9, P < 0.0001) for the neutralization of only 100 pg/ml IFN-α2 and/or -ω, and 20.8 (95% CI: 4.5–97.4, P < 0.0001) for the neutralization of 10 ng/ml IFN-α2 and -ω. Auto-Abs neutralizing type I IFNs accounted for ∼10% of severe TBE cases in these three European cohorts.

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Identification of Host Factors Differentially Induced by Clinically Diverse Strains of Tick-Borne Encephalitis Virus

Abstract: Tick-borne encephalitis virus (TBEV) is an emerging virus of the flavivirus family that is spread by ticks and causes neurological disease of various severity. No specific therapeutic treatments are available for TBE, and control in areas of endemicity is limited to vaccination.

Cited by 6 publications

References 73 publications

Functional characterization of 5′ untranslated region (UTR) secondary RNA structures in the replication of tick-borne encephalitis virus in mammalian cells

Functional characterization of 5′ untranslated region (UTR) secondary RNA structures in the replication of tick-borne encephalitis virus in mammalian cells

Activation of Early Proinflammatory Responses by TBEV NS1 Varies between the Strains of Various Subtypes

Autoantibodies neutralizing type I IFNs underlie severe tick-borne encephalitis in ∼10% of patients

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