A shared structural solution for neutralizing ebolaviruses

Dias, João M.; Kuehne, Ana I.; Abelson, Dafna M.; Bale, Shridhar; Wong, Anthony; Halfmann, Peter; Muhammad, Majidat; Fusco, Marnie L.; Zak, Samantha E.; Kang, Ellen S.; Kawaoka, Yoshihiro; Chandran, Kartik; Dye, John M.; Saphire, Erica Ollmann

doi:10.1038/nsmb.2150

Cited by 114 publications

(211 citation statements)

References 20 publications

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“…Ebolavirus proteins contain a significant fraction (20%) of structurally disordered regions, and the fraction of variable positions in these regions is significantly higher (p < 0.01) than in the structurally ordered regions. The 3D structures of globular regions are mostly known (Table S2) [54][55][56][57][58][59][60][61][62][63][64][65][66][67][68][69][70][71] except for the N-terminal zinc-finger domain of VP30, the coiled-coil domain of VP35, and protein L. Identification and analysis of structurally characterized homologs allowed us to predict the structure of the zinc-finger domain in VP30, the overall topology of NP, and the structure and catalytic sites for the catalytic domains of protein L.…”

Section: Resultsmentioning

confidence: 99%

Predictive and comparative analysis of Ebolavirus proteins

2015

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Ebolavirus is the pathogen for Ebola Hemorrhagic Fever (EHF). This disease exhibits a high fatality rate and has recently reached a historically epidemic proportion in West Africa. Out of the 5 known Ebolavirus species, only Reston ebolavirus has lost human pathogenicity, while retaining the ability to cause EHF in long-tailed macaque. Significant efforts have been spent to determine the three-dimensional (3D) structures of Ebolavirus proteins, to study their interaction with host proteins, and to identify the functional motifs in these viral proteins. Here, in light of these experimental results, we apply computational analysis to predict the 3D structures and functional sites for Ebolavirus protein domains with unknown structure, including a zinc-finger domain of VP30, the RNA-dependent RNA polymerase catalytic domain and a methyltransferase domain of protein L. In addition, we compare sequences of proteins that interact with Ebolavirus proteins from RESTV-resistant primates with those from RESTV-susceptible monkeys. The host proteins that interact with GP and VP35 show an elevated level of sequence divergence between the RESTV-resistant and RESTV-susceptible species, suggesting that they may be responsible for host specificity. Meanwhile, we detect variable positions in protein sequences that are likely associated with the loss of human pathogenicity in RESTV, map them onto the 3D structures and compare their positions to known functional sites. VP35 and VP30 are significantly enriched in these potential pathogenicity determinants and the clustering of such positions on the surfaces of VP35 and GP suggests possible uncharacterized interaction sites with host proteins that contribute to the virulence of Ebolavirus.

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Section: Resultsmentioning

confidence: 99%

Predictive and comparative analysis of Ebolavirus proteins

2015

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“…For the EBOV study, the fractionated IgG contained an endotoxin level of 7.54 EU/mg of protein and was assessed to be 96% pure by nonreduced and reduced SDS-PAGE. Plaque-reduction neutralization assay was performed as previously described (31).…”

Section: Methodsmentioning

confidence: 99%

Postexposure antibody prophylaxis protects nonhuman primates from filovirus disease

Dye

Herbert²,

Kuehne³

et al. 2012

Proc. Natl. Acad. Sci. U.S.A.

246

215

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Antibody therapies to prevent or limit filovirus infections have received modest interest in recent years, in part because of early negative experimental evidence. We have overcome the limitations of this approach, leveraging the use of antibody from nonhuman primates (NHPs) that survived challenge to filoviruses under controlled conditions. By using concentrated, polyclonal IgG antibody from these survivors, we treated filovirus-infected NHPs with multiple doses administered over the clinical phase of disease. In the first study, Marburg virus (MARV)-infected NHPs were treated 15 to 30 min postexposure with virus-specific IgG, with additional treatments on days 4 and 8 postexposure. The postexposure IgG treatment was completely protective, with no signs of disease or detectable viremia. MARV-specific IgM antibody responses were generated, and all macaques survived rechallenge with MARV, suggesting that they generated an immune response to virus replication. In the next set of studies, NHPs were infected with MARV or Ebola virus (EBOV), and treatments were delayed 48 h, with additional treatments on days 4 and 8 postexposure. The delayed treatments protected both MARV- and EBOV-challenged NHPs. In both studies, two of the three IgG-treated NHPs had no clinical signs of illness, with the third NHP developing mild and delayed signs of disease followed by full recovery. These studies clearly demonstrate that postexposure antibody treatments can protect NHPs and open avenues for filovirus therapies for human use using established Food and Drug Administration-approved polyclonal or monoclonal antibody technologies.

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“…To create viral antigen-specific assays, recombinant filovirus VP40 (rVP40) and NP (rNP) were prepared for antibody production and assay development [12][13][14][15]. Prior to the 2014 emergence of EVD in Guinea, ReEBOV RDT prototypes were developed using rVP40 and rNP antigens and respective polyclonal and monoclonal antibodies [16].…”

Section: Ebola Critical Reagent Developmentmentioning

confidence: 99%

Analytical Validation of the ReEBOV Antigen Rapid Test for Point-of-Care Diagnosis of Ebola Virus Infection

et al. 2016

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Background. Ebola virus disease (EVD) is a severe viral illness caused by Ebola virus (EBOV). The 2013-2016 EVD outbreak inWest Africa is the largest recorded, with >11 000 deaths. Development of the ReEBOV Antigen Rapid Test (ReEBOV RDT) was expedited to provide a point-of-care test for suspected EVD cases.Methods. Recombinant EBOV viral protein 40 antigen was used to derive polyclonal antibodies for RDT and enzyme-linked immunosorbent assay development. ReEBOV RDT limits of detection (LOD), specificity, and interference were analytically validated on the basis of Food and Drug Administration (FDA) guidance.Results. The ReEBOV RDT specificity estimate was 95% for donor serum panels and 97% for donor whole-blood specimens. The RDT demonstrated sensitivity to 3 species of Ebolavirus (Zaire ebolavirus, Sudan ebolavirus, and Bundibugyo ebolavirus) associated with human disease, with no cross-reactivity by pathogens associated with non-EBOV febrile illness, including malaria parasites. Interference testing exhibited no reactivity by medications in common use. The LOD for antigen was 4.7 ng/test in serum and 9.4 ng/test in whole blood. Quantitative reverse transcription-polymerase chain reaction testing of nonhuman primate samples determined the range to be equivalent to 3.0 × 10 5 -9.0 × 10 8 genomes/mL.Conclusions. The analytical validation presented here contributed to the ReEBOV RDT being the first antigen-based assay to receive FDA and World Health Organization emergency use authorization for this EVD outbreak, in February 2015.

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A shared structural solution for neutralizing ebolaviruses

Cited by 114 publications

References 20 publications

Predictive and comparative analysis of Ebolavirus proteins

Predictive and comparative analysis of Ebolavirus proteins

Postexposure antibody prophylaxis protects nonhuman primates from filovirus disease

Analytical Validation of the ReEBOV Antigen Rapid Test for Point-of-Care Diagnosis of Ebola Virus Infection

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