Poliovirus: Generation, Quantification, Propagation, Purification, and Storage

Burrill, Cecily P.; Strings, Vanessa R.; Andino, Raul

doi:10.1002/9780471729259.mc15h01s29

Cited by 23 publications

(34 citation statements)

References 15 publications

(21 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Plasmids were verified with Sanger sequencing for the introduced gatekeeper mutation. Protocols for virus generation from infectious cDNA clone and quantification have been previously described (Burrill et al, 2013a). …”

Section: Star Methods Textmentioning

confidence: 99%

The Evolutionary Pathway to Virulence of an RNA Virus

Stern

Yeh

Zinger

et al. 2017

Cell

Self Cite

120

177

View full text Add to dashboard Cite

Summary Paralytic polio once afflicted almost half a million children each year. The attenuated oral polio vaccine (OPV) has enabled world-wide vaccination efforts, which resulted in nearly complete control of the disease. However, poliovirus eradication is hampered globally by epidemics of vaccine-derived polio. Here, we describe a combined theoretical and experimental strategy that describes the molecular events leading from OPV to virulent strains. We discover that similar evolutionary events occur in most epidemics. The mutations and the evolutionary trajectories driving these epidemics are replicated using a simple cell-based experimental setup where the rate of evolution is intentionally accelerated. Furthermore, mutations accumulating during epidemics increase the replication fitness of the virus in cell culture and increase virulence in an animal model. Our study uncovers the evolutionary strategies by which vaccine strains become pathogenic, and provides a powerful framework for rational design of safer vaccine strains and for forecasting virulence of viruses.

show abstract

Section: Star Methods Textmentioning

confidence: 99%

The Evolutionary Pathway to Virulence of an RNA Virus

Stern

Yeh

Zinger

et al. 2017

Cell

Self Cite

120

177

View full text Add to dashboard Cite

show abstract

“…Poliovirus antibody assay. Available serum samples from infants collected after the third OPV vaccination were assessed for neutralizing antibodies against human poliovirus Sabin 1-3 serotypes, as previously described 23,24 . Poliovirus neutralization was assessed by plaque-reduction neutralization testing (PRNT) using replication-competent poliovirus propagated in HeLa S3 cells over a period of 7 days 20 .…”

Section: Nucleic Acid Extractionmentioning

confidence: 99%

Metagenomic sequencing of stool samples in Bangladeshi infants: virome association with poliovirus shedding after oral poliovirus vaccination

Granados

Bouquet

Hoy-Schulz

et al. 2020

Sci Rep

Self Cite

View full text Add to dashboard Cite

The potential role of enteric viral infections and the developing infant virome in affecting immune responses to the oral poliovirus vaccine (OPV) is unknown. Here we performed viral metagenomic sequencing on 3 serially collected stool samples from 30 Bangladeshi infants following OPV vaccination and compared findings to stool samples from 16 age-matched infants in the United States (US). In 14 Bangladeshi infants, available post-vaccination serum samples were tested for polio-neutralizing antibodies. The abundance (p = 0.006) and richness (p = 0.013) of the eukaryotic virome increased with age and were higher than seen in age-matched US infants (p < 0.001). In contrast, phage diversity metrics remained stable and were similar to those in US infants. Non-poliovirus eukaryotic virus abundance (3.68 log10 vs. 2.25 log10, p = 0.002), particularly from potential viral pathogens (2.78log10 vs. 0.83log10, p = 0.002), and richness (p = 0.016) were inversely associated with poliovirus shedding. Following vaccination, 28.6% of 14 infants tested developed neutralizing antibodies to all three Sabin types and also exhibited higher rates of poliovirus shedding (p = 0.020). No vaccine-derived poliovirus variants were detected. These results reveal an inverse association between eukaryotic virome abundance and poliovirus shedding. Overall gut virome ecology and concurrent viral infections may impact oral vaccine responsiveness in Bangladeshi infants.

show abstract

“…Three out of twenty-one clones exhibited the phenotype. The cDNA from these clones was subjected to PCR, and the entire viral genome was sequenced as described previously (28). A 3D4A P0 stock was subject to limiting dilution, and 16 clones were sequenced as described.…”

Section: Methodsmentioning

confidence: 99%

Global RNA Structure Analysis of Poliovirus Identifies a Conserved RNA Structure Involved in Viral Replication and Infectivity

Burrill

Westesson

Schulte

et al. 2013

J Virol

Self Cite

View full text Add to dashboard Cite

The genomes of RNA viruses often contain RNA structures that are crucial for translation and RNA replication and may play additional, uncharacterized roles during the viral replication cycle. For the picornavirus family member poliovirus, a number of functional RNA structures have been identified, but much of its genome, especially the open reading frame, has remained uncharacterized. We have now generated a global RNA structure map of the poliovirus genome using a chemical probing approach that interrogates RNA structure with single-nucleotide resolution. In combination with orthogonal evolutionary analyses, we uncover several conserved RNA structures in the open reading frame of the viral genome. To validate the ability of our global analyses to identify functionally important RNA structures, we further characterized one of the newly identified structures, located in the region encoding the RNA-dependent RNA polymerase, 3D pol , by site-directed mutagenesis. Our results reveal that the structure is required for viral replication and infectivity, since synonymous mutants are defective in these processes. Furthermore, these defects can be partially suppressed by mutations in the viral protein 3C pro , which suggests the existence of a novel functional interaction between an RNA structure in the 3D pol -coding region and the viral protein(s) 3C pro and/or its precursor 3CD pro .T he genomes of RNA viruses, such as poliovirus (PV), often contain complex RNA secondary and tertiary structures. These structures are crucial for translation and replication of the viral genome and may play additional roles in other processes, such as genome packaging and modulation of the host antiviral response.Poliovirus, the prototypical picornavirus and causative agent of poliomyelitis, is a nonenveloped virus with a single-stranded RNA genome of positive polarity. The virion consists of an icosahedral protein shell, composed of four capsid proteins (VP1, VP2, VP3, and VP4), which encapsidates the RNA genome (1). Poliovirus has a rapid replication cycle, with approximately 8 h elapsing between infection and release of progeny virions upon host cell lysis. During an infection, high yields of both viral proteins and genomes are produced. These yields ensure synthesis of up to 10,000 virions per cell (2), which can be widely disseminated to neighboring cells and/or new hosts. The compact nature of the viral genome (less than 7,500 nucleotides [nt] long) facilitates this rapid exponential growth. The viral genome acts as an mRNA and can be divided into a highly conserved 742-nt 5= untranslated region (5= UTR), a single long open reading frame encoding the viral polyprotein, a 68-nt 3= untranslated region (3= UTR), and a polyadenosine tract of a variable length (see Fig. 2A). A small viral protein of 22 amino acids, VPg (3B), is covalently attached to the 5= end of the RNA. The 5= UTR contains a structure critical for viral translation (the internal ribosome entry site or IRES) (3, 4), as well the 5=-cloverleaf (5=-CL) structure, which i...

show abstract

Poliovirus: Generation, Quantification, Propagation, Purification, and Storage

Cited by 23 publications

References 15 publications

The Evolutionary Pathway to Virulence of an RNA Virus

The Evolutionary Pathway to Virulence of an RNA Virus

Metagenomic sequencing of stool samples in Bangladeshi infants: virome association with poliovirus shedding after oral poliovirus vaccination

Global RNA Structure Analysis of Poliovirus Identifies a Conserved RNA Structure Involved in Viral Replication and Infectivity

Contact Info

Product

Resources

About