Correlation of RNA secondary structure and attenuation of Sabin vaccine strains of poliovirus in tissue culture

Macadam, Andrew; Ferguson, Geraldine; Burlison, J.; Stone, David M.; Skuce, Robin; Almond, Jeffrey W.; Minor, Philip D.

doi:10.1016/0042-6822(92)90565-7

Cited by 67 publications

(64 citation statements)

References 32 publications

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“…In both structures, the distal stem of IRES SLD V critically influences neuronal viral gene expression and particle propagation, most likely due to secondary structure formation interfering with efficient IRES function (22,35,51). This scenario suggests involvement of RNA-binding proteins with affinity for IRES secondary structural features.…”

Section: Discussionmentioning

confidence: 99%

“…SLD V mutations were proposed to influence neurovirulence through their effect on IRES secondary structure (21,35,51). This is underscored by the isolation of PV1(S) revertants with increased neurovirulence that acquired a transition at nt 525 (U3C), restoring the stem structure disrupted by the attenuating nt 480 (G) mutation (8).…”

mentioning

confidence: 99%

“…Similarly, reversion of IRES SLD V mutations at nt 480 (G3A) of PV1(S) and nt 481 (G3A) of PV2(S) occurred in patients with VAPP (20), with corresponding effects on neurovirulence in nonhuman primates (29,34). SLD V mutations were proposed to influence neurovirulence through their effect on IRES secondary structure (21,35,51). This is underscored by the isolation of PV1(S) revertants with increased neurovirulence that acquired a transition at nt 525 (U3C), restoring the stem structure disrupted by the attenuating nt 480 (G) mutation (8).…”

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confidence: 99%

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Genetic Determinants of Cell Type-Specific Poliovirus Propagation in HEK 293 Cells

Campbell

Lin

Dobrikova

et al. 2005

J Virol

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The ability of poliovirus to propagate in neuronal cells can be reduced by introducing appropriate nucleotide substitutions into the viral genome. Specific mutations scattered throughout the poliovirus genome yielded the live attenuated vaccine strains of poliovirus. Neuron-specific propagation deficits of the Sabin strains are partially encrypted within a confined region of the internal ribosomal entry site (IRES), which carries attenuating point mutations in all three serotypes. Recently, high levels of neurovirulence attenuation were achieved with genetically engineered polioviruses containing heterologous IRES elements. This is exemplified with poliovirus recombinants replicating under control of a human rhinovirus type 2 (HRV2) IRES element. We have carried out experiments delineating the genetic basis for neuronal IRES function. Neuronal dysfunction of the HRV2 IRES is determined mainly by IRES stem-loop domain V, the locus for attenuating point mutations within the Sabin strains. Neuronal incompetence associated with HRV2 IRES domain V is substantially more pronounced than that observed with the attenuating IRES point mutation of the Sabin serotype 1 vaccine strain. Mix-and-match recombination of polio and HRV2 IRES domain V suggests that the attenuation phenotype correlates with overall structural features rather than primary sequence. Our experiments have identified HEK 293 cells as a novel system for the study of neuron-specific replication phenotypes of poliovirus. This cell line, originally derived from embryonic human kidney, has recently been described to display neuronal characteristics. We report propagation properties in HEK 293 cells for poliovirus recombinants with attenuated neurovirulence in experimental animals that corroborate this observation.Poliovirus (PV), a positive-strand RNA virus of approximately 7,500 nucleotides (nt), is the prototypic member of the genus Enterovirus in the family Picornaviridae. PV is the causative agent of poliomyelitis, a paralytic condition complicating ϳ1% of all PV infections. The incidence of poliomyelitis has been drastically reduced since the introduction of two effective vaccines-the live attenuated (Sabin) strains and the formalininactivated (Salk) vaccine. Serial passage of wild-type (wt) PV in various culture systems yielded the serotype 1 and 3 Sabin strains, while serotype 2 is a naturally occurring variant with inherently reduced neurovirulence (48).It is generally assumed that the excellent safety record of the Sabin strains (the rate of vaccine-associated paralytic poliomyelitis [VAPP] is estimated at 0.14 per 1,000,000 doses) (60) is a direct reflection of tissue type-specific deficiencies that affect their propagation in the human central nervous system (CNS). Nevertheless, despite over 40 years of successful application, the molecular basis for neuroattenuation of the Sabin strains is not completely understood (16).Elucidation of the genomic sequences of PV type 1 (Sabin) [PV1(S)] (38) and PV3(S) (53, 58) and comparison to their wt progenitor...

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Section: Discussionmentioning

confidence: 99%

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confidence: 99%

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confidence: 99%

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Genetic Determinants of Cell Type-Specific Poliovirus Propagation in HEK 293 Cells

Campbell

Lin

Dobrikova

et al. 2005

J Virol

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“…Derivation of the hybrid type 3/type 2 virus Leon/Lansing (L3L2) and construction of L3L2 472/537 UG and L3L2 472/537 UA have been described previously (20,38). All mutants listed in Table 2 were constructed in a similar way.…”

Section: Methodsmentioning

confidence: 99%

“…BGM cells were used for temperature sensitivity assays as previously described (22). L20B cells were used for temperature sensitivity assays as described previously for CM-1 cells (20). Briefly, viruses were assayed by plaque formation at different temperatures.…”

Section: Methodsmentioning

confidence: 99%

Rational Design of Genetically Stable, Live-Attenuated Poliovirus Vaccines of All Three Serotypes: Relevance to Poliomyelitis Eradication

Macadam

Ferguson

Stone

et al. 2006

J Virol

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The global eradication of poliomyelitis caused by wild-type virus is likely to be completed within the next few years, despite immense logistic and political difficulties, and may ultimately be followed by the cessation of vaccination. However, the existing live-attenuated vaccines have the potential to revert to virulence, causing occasional disease, and viruses can be shed by immunocompromised individuals for prolonged periods of time. Moreover, several outbreaks of poliomyelitis have been shown to be caused by viruses derived from the Sabin vaccine strains. The appearance of such strains depends on the prevailing circumstances but poses a severe obstacle to strategies for stopping vaccination. Vaccine strains that are incapable of reversion at a measurable rate would provide a possible solution. Here, we describe the constructions of strains of type 3 poliovirus that are stabilized by the introduction of four mutations in the 5 noncoding region compared to the present vaccine. The strains are genetically and phenotypically stable under conditions where the present vaccine loses the attenuating mutation in the 5 noncoding region completely. Type 1 and type 2 strains in which the entire 5 noncoding regions of Sabin 1 and Sabin 2 were replaced exactly with that of one of the type 3 strains were also constructed. The genetic stability of 5 noncoding regions of these viruses matched that of the type 3 strains, but significant phenotypic reversion occurred, illustrating the potential limitations of a rational approach to the genetic stabilization of live RNA virus vaccines.Poliomyelitis is caused by poliovirus, a member of the picornavirus family that occurs in three serotypes such that immunity to one serotype does not protect against the other two.

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Role for poliovirus protease 2A in cap independent translation.

et al. 1994

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Viral protein synthesis in poliovirus infected cells was found to be influenced by mutations in part of the viral 5′‐non‐coding region (NCR) in a temperature dependent manner. At elevated temperatures these mutations resulted in virus titre reductions that allowed selection of revertant viruses. Some revertants were found to have retained the 5′‐NCR mutations but had compensating mutations in the 2A protease gene that were responsible for the suppression of the temperature sensitive phenotypes. The mutations in 2A enhanced viral protein synthesis at a stage when cap dependent translation was already abolished, suggesting that the virally encoded protein 2A is directly involved in the process of cap independent translation in addition to its role in abolishing cap dependent translation.

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Correlation of RNA secondary structure and attenuation of Sabin vaccine strains of poliovirus in tissue culture

Cited by 67 publications

References 32 publications

Genetic Determinants of Cell Type-Specific Poliovirus Propagation in HEK 293 Cells

Genetic Determinants of Cell Type-Specific Poliovirus Propagation in HEK 293 Cells

Rational Design of Genetically Stable, Live-Attenuated Poliovirus Vaccines of All Three Serotypes: Relevance to Poliomyelitis Eradication

Role for poliovirus protease 2A in cap independent translation.

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