scholarly journals Envelope Protein Glycosylation Mediates Zika Virus Pathogenesis

2019 ◽  
Vol 93 (12) ◽  
Author(s):  
Derek L. Carbaugh ◽  
Ralph S. Baric ◽  
Helen M. Lazear
Keyword(s):  
Zika Virus ◽  
Envelope Protein ◽  
Glycosylation Site ◽  
Viral Envelope ◽  
Protein Glycosylation ◽  
Guillain Barre Syndrome ◽  
Viral Loads ◽  
E Protein ◽  
Guillain Barré Syndrome ◽  
Guillain Barré

ABSTRACT Zika virus (ZIKV) is an emerging mosquito-borne flavivirus. Recent ZIKV outbreaks have produced serious human disease, including neurodevelopmental malformations (congenital Zika syndrome) and Guillain-Barré syndrome. These outcomes were not associated with ZIKV infection prior to 2013, raising the possibility that viral genetic changes could contribute to new clinical manifestations. All contemporary ZIKV isolates encode an N-linked glycosylation site in the envelope (E) protein (N154), but this glycosylation site is absent in many historical ZIKV isolates. Here, we investigated the role of E protein glycosylation in ZIKV pathogenesis using two contemporary Asian-lineage strains (H/PF/2013 and PRVABC59) and the historical African-lineage strain (MR766). We found that glycosylated viruses were highly pathogenic in Ifnar1−/− mice. In contrast, nonglycosylated viruses were attenuated, producing lower viral loads in the serum and brain when inoculated subcutaneously but remaining neurovirulent when inoculated intracranially. These results suggest that E glycosylation is advantageous in the periphery but not within the brain. Accordingly, we found that glycosylation facilitated infection of cells expressing the lectins dendritic cell-specific intercellular adhesion molecule-3-grabbing nonintegrin (DC-SIGN) or DC-SIGN-related (DC-SIGNR), suggesting that inefficient infection of lectin-expressing leukocytes could contribute to the attenuation of nonglycosylated ZIKV in mice. IMPORTANCE It is unclear why the ability of Zika virus (ZIKV) to cause serious disease, including Guillain-Barré syndrome and birth defects, was not recognized until recent outbreaks. One contributing factor could be genetic differences between contemporary ZIKV strains and historical ZIKV strains. All isolates from recent outbreaks encode a viral envelope protein that is glycosylated, whereas many historical ZIKV strains lack this glycosylation. We generated nonglycosylated ZIKV mutants from contemporary and historical strains and evaluated their virulence in mice. We found that nonglycosylated viruses were attenuated and produced lower viral loads in serum and brains. Our studies suggest that envelope protein glycosylation contributes to ZIKV pathogenesis, possibly by facilitating attachment to and infection of lectin-expressing leukocytes.

scholarly journals Zika Virus Encoding Nonglycosylated Envelope Protein Is Attenuated and Defective in Neuroinvasion

2017 ◽  
Vol 91 (23) ◽  
Author(s):  
Arun S. Annamalai ◽  
Aryamav Pattnaik ◽  
Bikash R. Sahoo ◽  
Ezhumalai Muthukrishnan ◽  
Sathish Kumar Natarajan ◽  
...  
Keyword(s):  
Mouse Model ◽  
Zika Virus ◽  
Glycosylation Site ◽  
Human Diseases ◽  
Guillain Barre Syndrome ◽  
Sequence Motif ◽  
Genetic Changes ◽  
E Protein ◽  
Guillain Barré Syndrome ◽  
Guillain Barré

ABSTRACT Zika virus (ZIKV), a mosquito-transmitted flavivirus responsible for sporadic outbreaks of mild and febrile illness in Africa and Asia, reemerged in the last decade causing serious human diseases, including microcephaly, congenital malformations, and Guillain-Barré syndrome. Although genomic and phylogenetic analyses suggest that genetic evolution may have led to the enhanced virulence of ZIKV, experimental evidence supporting the role of specific genetic changes in virulence is currently lacking. One sequence motif, VNDT, containing an N-linked glycosylation site in the envelope (E) protein, is polymorphic; it is absent in many of the African isolates but present in all isolates from the recent outbreaks. In the present study, we investigated the roles of this sequence motif and glycosylation of the E protein in the pathogenicity of ZIKV. We first constructed a stable full-length cDNA clone of ZIKV in a novel linear vector from which infectious virus was recovered. The recombinant ZIKV generated from the infectious clone, which contains the VNDT motif, is highly pathogenic and causes lethality in a mouse model. In contrast, recombinant viruses from which the VNDT motif is deleted or in which the N-linked glycosylation site is mutated by single-amino-acid substitution are highly attenuated and nonlethal. The mutant viruses replicate poorly in the brains of infected mice when inoculated subcutaneously but replicate well following intracranial inoculation. Our findings provide the first evidence that N-linked glycosylation of the E protein is an important determinant of ZIKV virulence and neuroinvasion. IMPORTANCE The recent emergence of Zika virus (ZIKV) in the Americas has caused major worldwide public health concern. The virus appears to have gained significant pathogenicity, causing serious human diseases, including microcephaly and Guillain-Barré syndrome. The factors responsible for the emergence of pathogenic ZIKV are not understood at this time, although genetic changes have been shown to facilitate virus transmission. All isolates from the recent outbreaks contain an N-linked glycosylation site within the viral envelope (E) protein, whereas many isolates of the African lineage virus lack this site. To elucidate the functional significance of glycosylation in ZIKV pathogenicity, recombinant ZIKVs from infectious clones with or without the glycan on the E protein were generated. ZIKVs lacking the glycan were highly attenuated for the ability to cause mortality in a mouse model and were severely compromised for neuroinvasion. Our studies suggest glycosylation of the E protein is an important factor contributing to ZIKV pathogenicity.

scholarly journals Isolation of infective Zika virus from urine and saliva of patients in Brazil

2016 ◽  
Author(s):  
Myrna C. Bonaldo ◽  
Ieda P. Ribeiro ◽  
Noemia S. Lima ◽  
Alexandre A. C. dos Santos ◽  
Lidiane S. R. Menezes ◽  
...  
Keyword(s):  
Acute Phase ◽  
Zika Virus ◽  
Sexual Transmission ◽  
Critical Factor ◽  
Transmission Efficiency ◽  
Guillain Barre Syndrome ◽  
Molecular Diagnostic ◽  
Viral Loads ◽  
Guillain Barré Syndrome ◽  
Guillain Barré

ABSTRACTBACKGROUNDZika virus (ZIKV) is an emergent threat provoking a worldwide explosive outbreak. Since January 2015, 41 countries reported autochthonous cases. In Brazil, an increase in Guillain-Barré syndrome and microcephaly cases was linked to ZIKV infections. A recent report describing low experimental transmission efficiency of its main putative vector, Ae. aegypti, in conjunction with apparent sexual transmission notifications prompted the investigation of other potential sources of viral dissemination. Urine and saliva have been previously established as useful tools in ZIKV diagnosis. However, no evidence regarding the infectivity of ZIKV particles present in saliva and urine has been obtained yet.METHODOLOGY/PRINCIPAL FINDINGSNine urine and five saliva samples from nine patients from Rio de Janeiro presenting rash and other typical Zika acute phase symptoms were inoculated in Vero cell culture and submitted to specific ZIKV RNA detection and quantification through, respectively, NAT-Zika, RT-PCR and RT-qPCR. Two ZIKV isolates were achieved, one from urine and one from saliva specimens. ZIKV nucleic acid was identified by all methods in four patients. Whenever both urine and saliva samples were available from the same patient, urine viral loads were higher, corroborating the general sense that it is a better source for ZIKV molecular diagnostic. In spite of this, from the two isolated strains, each from one patient, only one derived from urine, suggesting that other factors, like the acidic nature of this fluid, might interfere with virion infectivity. The complete genome of both ZIKV isolates was obtained. Phylogenetic analysis revealed similarity with strains previously isolated during the South America outbreak.CONCLUSIONS/SIGNIFICANCEThe detection of infectious ZIKV particles in urine and saliva of patients during the acute phase may represent a critical factor in the spread of virus. The epidemiological relevance of this finding, regarding the contribution of alternative non vectorial ZIKV transmission routes, needs further investigation.AUTHOR SUMMARYThe American continent has recently been the scene of a devastating epidemic of Zika virus and its severe manifestations, such as microcephaly in newborns and Guillain-Barré Syndrome. Zika virus, first detected in 1947 in Africa, only from 2007 started provoking outbreaks. Zika, dengue and chikungunya viruses are primarily transmitted by Aedes mosquitoes. Dengue is endemic in Brazil for almost 30 years, and the country is largely infested by its main vector, Aedes aegypti. Chikungunya virus entered the country in late 2014 and Zika presence was confirmed eight months later. Nevertheless, Zika notifications multiplied and spread across the country with unprecedented speed, raising the possibility of other transmission routes. This hypothesis was strengthened by some recent reports of Zika sexual transmission in Ae. aegypti-free areas and by the description of a low transmission efficiency to Zika virus in local Ae. aegypti. We found Zika active particles in both urine and saliva of acute phase patients, and a finding that was promptly announced by Fiocruz via Press Conference on February 5, 2016. In this work, we bring up the potential alternative person-to-person infection routes beyond the vectorial transmission, that might have epidemiological relevance.

scholarly journals Zika E Glycan Loop Region and Guillain–Barré Syndrome-Related Proteins: A Possible Molecular Mimicry to Be Taken in Account for Vaccine Development

Vaccines ◽  
2021 ◽  
Vol 9 (3) ◽  
pp. 283
Author(s):  
Grégorie Lebeau ◽  
Etienne Frumence ◽  
Jonathan Turpin ◽  
Floran Begue ◽  
Jean-Jacques Hoarau ◽  
...  
Keyword(s):  
Heat Shock ◽  
Autoimmune Diseases ◽  
Vaccine Development ◽  
Molecular Mimicry ◽  
Viral Envelope ◽  
Guillain Barre Syndrome ◽  
Loop Region ◽  
E Protein ◽  
Guillain Barré Syndrome ◽  
Guillain Barré

The neurological complications of infection by the mosquito-borne Zika virus (ZIKV) include Guillain–Barré syndrome (GBS), an acute inflammatory demyelinating polyneuritis. GBS was first associated with recent ZIKV epidemics caused by the emergence of the ZIKV Asian lineage in South Pacific. Here, we hypothesize that ZIKV-associated GBS relates to a molecular mimicry between viral envelope E (E) protein and neural proteins involved in GBS. The analysis of the ZIKV epidemic strains showed that the glycan loop (GL) region of the E protein includes an IVNDT motif which is conserved in voltage-dependent L-type calcium channel subunit alpha-1C (Cav1.2) and Heat Shock 70 kDa protein 12A (HSP70 12A). Both VSCC-alpha 1C and HSP70 12A belong to protein families which have been associated with neurological autoimmune diseases in central nervous system. The purpose of our in silico analysis is to point out that IVNDT motif of ZIKV E-GL region should be taken in consideration for the development of safe and effective anti-Zika vaccines by precluding the possibility of adverse neurologic events including autoimmune diseases such as GBS through a potent mimicry with Heat Shock 70 kDa protein 12A (HSP70 12A).

scholarly journals Site-specific N-glycosylation analysis of animal cell culture-derived Zika virus proteins

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Alexander Pralow ◽  
Alexander Nikolay ◽  
Arnaud Leon ◽  
Yvonne Genzel ◽  
Erdmann Rapp ◽  
...  
Keyword(s):  
Zika Virus ◽  
Animal Cell ◽  
Gradient Centrifugation ◽  
Viral Envelope ◽  
Protein Glycosylation ◽  
High Yield ◽  
Structural Protein ◽  
Site Specific ◽  
E Protein ◽  
The Impact

AbstractHere, we present for the first time, a site-specific N-glycosylation analysis of proteins from a Brazilian Zika virus (ZIKV) strain. The virus was propagated with high yield in an embryo-derived stem cell line (EB66, Valneva SE), and concentrated by g-force step-gradient centrifugation. Subsequently, the sample was proteolytically digested with different enzymes, measured via a LC–MS/MS-based workflow, and analyzed in a semi-automated way using the in-house developed glyXtoolMS software. The viral non-structural protein 1 (NS1) was glycosylated exclusively with high-mannose structures on both potential N-glycosylation sites. In case of the viral envelope (E) protein, no specific N-glycans could be identified with this method. Nevertheless, N-glycosylation could be proved by enzymatic de-N-glycosylation with PNGase F, resulting in a strong MS-signal of the former glycopeptide with deamidated asparagine at the potential N-glycosylation site N444. This confirmed that this site of the ZIKV E protein is highly N-glycosylated but with very high micro-heterogeneity. Our study clearly demonstrates the progress made towards site-specific N-glycosylation analysis of viral proteins, i.e. for Brazilian ZIKV. It allows to better characterize viral isolates, and to monitor glycosylation of major antigens. The method established can be applied for detailed studies regarding the impact of protein glycosylation on antigenicity and human pathogenicity of many viruses including influenza virus, HIV and corona virus.

scholarly journals Zika Virus and the Guillain–Barré Syndrome — Case Series from Seven Countries

2016 ◽  
Vol 375 (16) ◽  
pp. 1598-1601 ◽  
Author(s):  
Thais dos Santos ◽  
Angel Rodriguez ◽  
Maria Almiron ◽  
Antonio Sanhueza ◽  
Pilar Ramon ◽  
...  
Keyword(s):  
Zika Virus ◽  
Case Series ◽  
Guillain Barre Syndrome ◽  
Guillain Barré Syndrome ◽  
Guillain Barré

scholarly journals Zika Virus Infection as a Cause of Congenital Brain Abnormalities and Guillain–Barré Syndrome: Systematic Review

PLoS Medicine ◽  
2017 ◽  
Vol 14 (1) ◽  
pp. e1002203 ◽  
Author(s):  
Fabienne Krauer ◽  
Maurane Riesen ◽  
Ludovic Reveiz ◽  
Olufemi T. Oladapo ◽  
Ruth Martínez-Vega ◽  
...  
Keyword(s):  
Systematic Review ◽  
Virus Infection ◽  
Zika Virus ◽  
Guillain Barre Syndrome ◽  
Brain Abnormalities ◽  
Zika Virus Infection ◽  
Guillain Barré Syndrome ◽  
Guillain Barré

scholarly journals The re-emergence of Zika in Brazil in 2020: a case of Guillain Barré Syndrome during the low season for arboviral infections

2020 ◽  
Vol 27 (7) ◽  
Author(s):  
Kevan M Akrami ◽  
Betania Mara Freitas de Nogueira ◽  
Mateus Santana do Rosário ◽  
Laise de Moraes ◽  
Marli Tenório Cordeiro ◽  
...  
Keyword(s):  
Zika Virus ◽  
Guillain Barre Syndrome ◽  
Guillain Barré Syndrome ◽  
Guillain Barré

Zika virus cases in Brazil have diminished since emergence in 2015. We report Guillain Barré Syndrome caused by Zika and possible Chikungunya co-infection during an expected low arboviral season. This case highlights the importance of clinical vigilance for Zika in those with neurological syndromes outside typical arboviral season.

scholarly journals Zika virus infection as a cause of congenital brain abnormalities and Guillain-Barré syndrome: From systematic review to living systematic review

F1000Research ◽  
2018 ◽  
Vol 7 ◽  
pp. 196 ◽  
Author(s):  
Michel Jacques Counotte ◽  
Dianne Egli-Gany ◽  
Maurane Riesen ◽  
Million Abraha ◽  
Teegwendé Valérie Porgo ◽  
...  
Keyword(s):  
Systematic Review ◽  
Zika Virus ◽  
Congenital Abnormalities ◽  
Causal Link ◽  
The Body ◽  
Guillain Barre Syndrome ◽  
Sufficient Evidence ◽  
Zikv Infection ◽  
Guillain Barré Syndrome ◽  
Guillain Barré

Background. The Zika virus (ZIKV) outbreak in the Americas has caused international concern due to neurological sequelae linked to the infection, such as microcephaly and Guillain-Barré syndrome (GBS). The World Health Organization stated that there is “sufficient evidence to conclude that Zika virus is a cause of congenital abnormalities and is a trigger of GBS”. This conclusion was based on a systematic review of the evidence published until 30.05.2016. Since then, the body of evidence has grown substantially, leading to this update of that systematic review with new evidence published from 30.05.2016 – 18.01.2017, update 1. Methods. We review evidence on the causal link between ZIKV infection and adverse congenital outcomes and the causal link between ZIKV infection and GBS or immune-mediated thrombocytopaenia purpura. We also describe the transition of the review into a living systematic review, a review that is continually updated. Results. Between 30.05.2016 and 18.01.2017, we identified 2413 publications, of which 101 publications were included. The evidence added in this update confirms the conclusion of a causal association between ZIKV and adverse congenital outcomes. New findings expand the evidence base in the dimensions of biological plausibility, strength of association, animal experiments and specificity. For GBS, the body of evidence has grown during the search period for update 1, but only for dimensions that were already populated in the previous version. There is still a limited understanding of the biological pathways that potentially cause the occurrence of autoimmune disease following ZIKV infection. Conclusions. This systematic review confirms previous conclusions that ZIKV is a cause of congenital abnormalities, including microcephaly, and is a trigger of GBS. The transition to living systematic review techniques and methodology provides a proof of concept for the use of these methods to synthesise evidence about an emerging pathogen such as ZIKV.

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