Quantifying Antigenic Relationships among the Lyssaviruses

Horton, Daniel L.; McElhinney, L. M.; Marston, Denise A.; Wood, James; Russell, Colin A.; Lewis, Nicola S.; Kuzmin, Ivan V.; Fouchier, Ron A. M.; Osterhaus, Albert D. M. E.; Smith, Derek J.

doi:10.1128/jvi.01153-10

Cited by 80 publications

(108 citation statements)

References 48 publications

(57 reference statements)

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“…Indeed, for RV20 the total amount of RNA was too low to obtain 200 ng RNA for fragmentation. The virus titer of RV1787 and RV20 has been calculated previously [23,24] with RV1787 (EBLV-2) approximately 1 log lower than RV20, therefore the difference in the percentage of viral reads is likely to be a reflection of this. Despite the marked difference between the percentage of viral reads of RV20 and RV1787, the difference within samples regarding whether the RNA was depleted or not, is not so obvious.…”

Section: Resultsmentioning

confidence: 99%

Next generation sequencing of viral RNA genomes

et al. 2013

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BackgroundWith the advent of Next Generation Sequencing (NGS) technologies, the ability to generate large amounts of sequence data has revolutionized the genomics field. Most RNA viruses have relatively small genomes in comparison to other organisms and as such, would appear to be an obvious success story for the use of NGS technologies. However, due to the relatively low abundance of viral RNA in relation to host RNA, RNA viruses have proved relatively difficult to sequence using NGS technologies. Here we detail a simple, robust methodology, without the use of ultra-centrifugation, filtration or viral enrichment protocols, to prepare RNA from diagnostic clinical tissue samples, cell monolayers and tissue culture supernatant, for subsequent sequencing on the Roche 454 platform.ResultsAs representative RNA viruses, full genome sequence was successfully obtained from known lyssaviruses belonging to recognized species and a novel lyssavirus species using these protocols and assembling the reads using de novo algorithms. Furthermore, genome sequences were generated from considerably less than 200 ng RNA, indicating that manufacturers’ minimum template guidance is conservative. In addition to obtaining genome consensus sequence, a high proportion of SNPs (Single Nucleotide Polymorphisms) were identified in the majority of samples analyzed.ConclusionsThe approaches reported clearly facilitate successful full genome lyssavirus sequencing and can be universally applied to discovering and obtaining consensus genome sequences of RNA viruses from a variety of sources.

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Section: Resultsmentioning

confidence: 99%

Next generation sequencing of viral RNA genomes

et al. 2013

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“…Of the 15 lyssavirus species, RABV, EBLV-1 and -2, ABLV, DUVV, IRKV (all phylogroup 1) and MOKV, (phylogroup 2) have caused human deaths 1,15-20 . Studies have been conducted investigating cross-neutralization between sera against divergent members of the lyssavirus genus 21,22 . Human rabies vaccines, which contain inactivated RABV strains, have been shown to elicit adequate rabies virus neutralizing antibody (VNA) concentrations, not only against the classical RABV but also against other species of phylogroup 1, i.e., EBLV-1 and -2 and ABLV 23-25 .…”

Section: Introductionmentioning

confidence: 99%

Cross-neutralization of antibodies induced by vaccination with Purified Chick Embryo Cell Vaccine (PCECV) against differentLyssavirusspecies

Malerczyk

Freuling

Gniel

et al. 2014

Human Vaccines & Immunotherapeutics

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Background: Rabies is a neglected zoonotic disease caused by viruses belonging to the genus lyssavirus. In endemic countries of Asia and Africa, where the majority of the estimated 60,000 human rabies deaths occur, it is mainly caused by the classical rabies virus (RABV) transmitted by dogs. Over the last decade new species within the genus lyssavirus have been identified. Meanwhile 15 (proposed or classified) species exist, including Australian bat lyssavirus (ABLV), European bat lyssavirus (EBLV-1 and -2), Duvenhage virus (DUVV), as well as Lagos bat virus (LBV) and Mokola virus (MOKV) and recently identified novel species like Bokeloh bat lyssavirus (BBLV), Ikoma bat lyssavirus (IKOV) or Lleida bat lyssavirus (LLBV). The majority of these lyssavirus species are found in bat reservoirs and some have caused human infection and deaths. Previous work has demonstrated that Purified Chick Embryo Cell Rabies Vaccine (PCECV) not only induces immune responses against classical RABV, but also elicits cross-neutralizing antibodies against ABLV, EBLV-1 and EBLV-2.Material & Methods: Using the same serum samples as in our previous study, this study extension investigated cross-neutralizing activities of serum antibodies measured by rapid fluorescent focus inhibition test (RFFIT) against selected other non-classical lyssavirus species of interest, namely DUVV and BBLV, as well as MOKV and LBV.Results: Antibodies developed after vaccination with PCECV have neutralizing capability against BBLV and DUVV in the same range as against ABLV and EBLV-1 and -2. As expected, for the phylogenetically more distant species LBV no cross-neutralizing activity was found. Interestingly, 15 of 94 serum samples (16%) with a positive neutralizing antibody titer against RABV displayed specific cross-neutralizing activity (65-fold lower than against RABV) against one specific MOKV strain (Ethiopia isolate), which was not seen against a different strain (Nigeria isolate).Conclusion: Cross-neutralizing activities partly correlate with the phylogenetic distance of the virus species. Cross-neutralizing activities against the species BBLV and DUVV of phylogroup 1 were demonstrated, in line with previous results of cross-neutralizing activities against ABLV and EBLV-1 and -2. Potential partial cross-neutralizing activities against more distant lyssavirus species like selected MOKV strains need further research.

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“…This is particularly important as several virus-neutralizing monoclonal antibodies (MAbs) were offered for replacement of conventional HRIG in human rabies PEP (21)(22)(23)(24)(25)(26)(27)(28). For example, a linear epitope with the key residues LCGV within antigenic site I serves for binding of MAbs CR57 and 62071-3 (22,23).…”

Section: Discussionmentioning

confidence: 99%