Amino Acid Variation at VP1-145 of Enterovirus 71 Determines Attachment Receptor Usage and Neurovirulence in Human Scavenger Receptor B2 Transgenic Mice

Kobayashi, Kanae; Sudaka, Yui; Takashino, Ayako; Imura, Ayumi; Fujii, Ken; Koike, Sachiko

doi:10.1128/jvi.00681-18

Cited by 47 publications

(87 citation statements)

References 49 publications

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“…Therefore, HS is not expressed at high levels in SCARB2-positive neuronal cells. The distributions of HS and SCARB2 in monkeys are similar to those in hSCARB2 tg mice (28), suggesting that the HS attachment receptor does not contribute to productive infection mediated by SCARB2 in neuronal cells in vivo. Production of neutralizing antibody against EV71.…”

mentioning

confidence: 78%

“…Using an hSCARB2 tg mouse model, Kobayashi et al (28) found that VP1-145G viruses bind to HS and that HS is expressed highly in cells that do not express SCARB2. HSs act as decoys that trap the VP1-145G viruses and prevent their spread in vivo.…”

Section: Discussionmentioning

confidence: 99%

“…Vero, RD-A, and COS7 cells were maintained in Dulbecco's modified Eagle's medium (DMEM) (Nissui) supplemented with 5% fetal bovine serum (FBS). Generation of RD-SCARB2 cells was as described in the accompanying paper (28). RD-SCARB2 cells were cultured in DMEM supplemented with 5% FBS and 1 g/ml puromycin.…”

Section: Methodsmentioning

confidence: 99%

“…Distribution of HS in cynomolgus monkeys. We hypothesized that HS acts as a decoy to specifically adsorb VP1-145G viruses (28). The results above suggested that this hypothesis is also true in cynomolgus monkeys.…”

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confidence: 87%

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VP1 Amino Acid Residue 145 of Enterovirus 71 Is a Key Residue for Its Receptor Attachment and Resistance to Neutralizing Antibody during Cynomolgus Monkey Infection

Fujii

Sudaka

Takashino

et al. 2018

J Virol

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Enterovirus 71 (EV71) is a causative agent of hand, foot, and mouth disease and sometimes causes severe or fatal neurological complications. The amino acid at VP1-145 determines virological characteristics of EV71. Viruses with glutamic acid (E) at VP1-145 (VP1-145E) are virulent in neonatal mice and transgenic mice expressing human scavenger receptor B2, whereas those with glutamine (Q) or glycine (G) are not. However, the contribution of this variation to pathogenesis in humans is not fully understood. We compared the virulence of VP1-145E and VP1-145G viruses of Isehara and C7/Osaka backgrounds in cynomolgus monkeys. VP1-145E, but not VP1-145G, viruses induced neurological symptoms. VP1-145E viruses were frequently detected in the tissues of infected monkeys. VP1-145G viruses were detected less frequently and disappeared quickly. Instead, mutants that had a G to E mutation at VP1-145 emerged, suggesting that VP1-145E viruses have a replication advantage in the monkeys. This is consistent with our hypothesis proposed in the accompanying paper that the VP1-145G virus is attenuated due to its adsorption by heparan sulfate. Monkeys infected with both viruses produced neutralizing antibodies before the onset of the disease. Interestingly, VP1-145E viruses were more resistant to neutralizing antibodies than VP1-145G viruses A small amount of neutralizing antibody raised in the early phase of infection may not be sufficient to block the dissemination of VP1-145E viruses. The different resistance of the VP1-145 variants to neutralizing antibodies may be one of the reasons for the difference in virulence. The contribution of VP1-145 variants in humans is not fully understood. In some reports, VP1-145G/Q viruses were more frequently isolated from severely affected than from mildly affected patients, suggesting that VP1-145G/Q viruses are more virulent. In the accompanying paper, we showed that VP1-145E viruses are more virulent than VP1-145G viruses in human SCARB2 transgenic mice. Heparan sulfate acts as a decoy to specifically trap the VP1-145G viruses and leads to abortive infection. Here, we demonstrated that VP1-145G was attenuated in cynomolgus monkeys, suggesting that this hypothesis is also true in a non-human primate model. VP1-145E viruses, but not VP1-145G viruses, were highly resistant to neutralizing antibodies. We propose the difference in resistance against neutralizing antibodies as another mechanism of EV71 virulence. In summary, VP1-145 contributes to virulence determination by controlling attachment receptor usage and antibody sensitivity.

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confidence: 78%

Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

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confidence: 87%

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VP1 Amino Acid Residue 145 of Enterovirus 71 Is a Key Residue for Its Receptor Attachment and Resistance to Neutralizing Antibody during Cynomolgus Monkey Infection

Fujii

Sudaka

Takashino

et al. 2018

J Virol

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“…This gives rise to a low viremia level with a substantial reduction of virulence in mice. Kobayashi and colleagues reported that EGK virus is less virulent compared to EEK due to the former harbouring the VP1-G145 residue, enabling the virus to adsorb more strongly to HS, resulting in attenuated virulence in SCARB2-expressing transgenic mice [56]. Our findings that a strong heparin-binding phenotype attenuates virulence is also observed in other viruses, including Sindbis virus [33], Venezuelan equine encephalitis virus [55], West Nile virus [30], yellow fever virus [31] and Japanese encephalitis virus [57].…”

Section: Discussionmentioning

confidence: 99%

Electrostatic interactions at the five-fold axis alter heparin-binding phenotype and drive EV-A71 virulence in mice

Tee

Tan

Yogarajah

et al. 2019

Preprint

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19Enterovirus A71 (EV-A71) causes hand, foot and mouth disease epidemics with neurological 20 complications and fatalities. However, the neuropathogenesis of EV-A71 remains poorly 21 understood. In mice, adaptation and virulence determinants have been mapped to mutations at 22 VP1-145, VP1-244 and VP2-149. We hypothesized that heparin-binding phenotype shapes 23 EV-A71 virulence in mice. We constructed six viruses with varying residues at VP1-98, VP1-24 145 (which are both heparin-binding determinants) and VP2-149 (based on the wild type 25 98E/145Q/149K, termed EQK) to generate KQK, KEK, EEK, EEI and EQI variants. We 26 demonstrated that the weak heparin-binder EEI was highly lethal in mice. The initially strong 27 heparin-binding EQI variant acquired an additional mutation VP1-K244E, which confers weak 28 heparin-binding phenotype resulting in elevated viremia and increased brain inflammation and 29 virus antigens in mice, with subsequent high virulence. EEI and EQI-K244E variants 30 inoculated into mice disseminated efficiently and displayed high viremia. Increasing 31 polymerase fidelity and impairing recombination of EQI attenuated virulence, suggesting the 32 importance of population diversity in EV-A71 pathogenesis in vivo. Combining in silico 33 docking and deep sequencing approaches, we inferred that virus population diversity is shaped 34 by electrostatic interactions at the five-fold axis of the virus surface. Electrostatic surface 35 charges facilitate virus adaptation by generating poor heparin-binding variants for better in vivo 36 dissemination in mice, likely due to reduced adsorption to heparin-rich peripheral tissues, 37 which ultimately results in increased neurovirulence. The dynamic switching from heparin-38 binding to weak heparin-binding phenotype in vivo explained the neurovirulence of EV-A71. 39 40 Author summary 41 Enterovirus A71 (EV-A71) is the primary cause of hand, foot and mouth disease, and it can 42 also infect the central nervous system and cause fatal outbreaks in young children. EV-A71 43 pathogenesis remains elusive. In this study, we demonstrated that EV-A71 variants with strong 44 affinity to heparan sulfate (heparin) have a growth advantage in tissue culture, but are 45 disadvantageous in vivo. When inoculated into mice, strong heparin-binding virus variants are 46 more likely to be adsorbed to peripheral tissues, resulting in impaired ability to disseminate 47 and being cleared from the bloodstream rapidly. The lower viremia level resulted in no 48 neuroinvasion. In contrast, weak heparin-binding variants show greater levels of viremia, 49 dissemination and subsequent neurovirulence in mice. We also provide evidence that the ability 50 of EV-A71 to bind heparin is mediated by electrostatic surface charges due to amino acids on 51 the virus capsid surface. In mice, EV-A71 undergoes adaptive mutation to acquire greater 52 negative surface charges, thus generating new virulent variants with weak heparin-binding 53 which allows greater viral spread. Our study underlines the...

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Recent Progress on Functional Genomics Research of Enterovirus 71

Wang

2018

Virol. Sin.

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Enterovirus 71 (EV71) is one of the main pathogens that causes hand-foot-and-mouth disease (HFMD). HFMD caused by EV71 infection is mostly self-limited; however, some infections can cause severe neurological diseases, such as aseptic meningitis, brain stem encephalitis, and even death. There are still no effective clinical drugs used for the prevention and treatment of HFMD. Studying EV71 protein function is essential for elucidating the EV71 replication process and developing anti-EV71 drugs and vaccines. In this review, we summarized the recent progress in the studies of EV71 noncoding regions (5 0 UTR and 3 0 UTR) and all structural and nonstructural proteins, especially the key motifs involving in viral infection, replication, and immune regulation. This review will promote our understanding of EV71 virus replication and pathogenesis, and will facilitate the development of novel drugs or vaccines to treat EV71.

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Amino Acid Variation at VP1-145 of Enterovirus 71 Determines Attachment Receptor Usage and Neurovirulence in Human Scavenger Receptor B2 Transgenic Mice

Cited by 47 publications

References 49 publications

VP1 Amino Acid Residue 145 of Enterovirus 71 Is a Key Residue for Its Receptor Attachment and Resistance to Neutralizing Antibody during Cynomolgus Monkey Infection

VP1 Amino Acid Residue 145 of Enterovirus 71 Is a Key Residue for Its Receptor Attachment and Resistance to Neutralizing Antibody during Cynomolgus Monkey Infection

Electrostatic interactions at the five-fold axis alter heparin-binding phenotype and drive EV-A71 virulence in mice

Recent Progress on Functional Genomics Research of Enterovirus 71

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