Mapping of mutations contributing to the temperature sensitivity of the Sabin 1 vaccine strain of poliovirus

Abstract

The temperature-sensitive and attenuated phenotypes of the Sabin type 1 vaccine strain of poliovirus result from numerous point mutations which occurred in the virulent Mahoney virus parent. One of these mutations is located in a 3D polymerase (3D pol ) codon (U-62033C, Tyr-733His) and is involved in attenuation in common mice (M. Tardy-Panit, B. Blondel, A. Martin, F. Tekaia, F. Horaud, and F. Delpeyroux, J. Virol. 67: [4630][4631][4632][4633][4634][4635][4636][4637][4638] 1993). This mutation also appears to contribute to temperature sensitivity, in association with at least 1 other of the 10 mutations of the 3-terminal part of the genome including the 3D pol coding and 3 noncoding regions. To map the other mutation(s), we constructed poliovirus mutants by mutagenesis and recombination of Mahoney and Sabin 1 cDNAs. Characterization of these poliovirus mutants showed that a second mutation in a 3D pol codon (C-70713U, Thr-3623Ile) contributes to temperature sensitivity. A mutation in the 3 noncoding region of the genome (A-74413G), alone or linked to another mutation (U-74103C), also appeared to be involved in this phenotype. The temperature-sensitive effect associated with the 3-terminal part of the Sabin 1 genome results from the cumulative and/or synergistic effects of at least three genetic determinants, i.e., the His-73 and Ile-362 codons of 3D pol and nucleotide G-7441. Sequence analysis of strains isolated from patients with vaccine-associated paralytic poliomyelitis showed that these genetic determinants are selected against in vivo, although the Ile-362 codon appeared to be more stable than either the His-73 codon or G-7441. These genetic determinants may contribute to the safety of Sabin 1 in vaccinees.Poliovirus (PV), an enterovirus of the Picornaviridae family, is the causative agent of poliomyelitis and is classified into three distinct serotypes. It replicates in the human digestive tract and may induce paralysis by infecting and destroying motor neurons (5, 10). Poliomyelitis has been effectively controlled by the use of inactivated or live attenuated vaccines. Attenuated strains of all three serotypes have been selected by numerous passages of wild-type strains in monkey tissues in vivo and in vitro (43). These strains (Sabin 1, 2, and 3), which replicate in the human gut and induce a strong immunity, have a good safety record. However, in a small number of cases, vaccination is associated with paralytic poliomyelitis (34). Vaccine-associated paralytic poliomyelitis (VAPP) may result from the genetic variability (point mutations and recombination) of the Sabin strains. Indeed, neurovirulent vaccine-derived strains are found in the gut of healthy vaccinees and in the central nervous system of patients with VAPP (16, 31). VAPP is most frequently associated with Sabin 2 and Sabin 3 and rarely with Sabin 1 (3, 37). Analysis of the attenuation determinants of Sabin 1 could lead to a better understanding of the safety of this strain in vaccinees.PV is composed of an icosahedral protein shell which cont...

Vaccine-derived Mutation in Motif D of Poliovirus RNA-dependent RNA Polymerase Lowers Nucleotide Incorporation Fidelity

“…This finding suggests that there may be communication pathways linking the Sabin sites, and modifications at these sites may act cooperatively to change RdRp function; previous MD simulations of WT RdRp have also indicated that the motions of the ␣-helix containing Tyr-73 are anti-correlated with the motions of motif D containing Thr-362 (42). The Sabin amino acid substitutions may also contribute to different RdRp functions; the Y73H substitution has been shown to interfere with RdRp initiation (62), K250E may interfere with protein structure and/or long-range interactions with RNA through changes to electrostatic interactions, and T362I, in addition to its role in RdRp fidelity shown here, has been shown to contribute to temperature sensitivity of the Sabin I vaccine (59). The other Sabin substitutions may even compensate for the effects observed here with T362I.…”

Genetic evolution of enterovirus 71: epidemiological and pathological implications

“…This is supported by the identification of a VP3-Phen91 mutation, near a promoter interface in poliovirus Sabin-3 strain, which affects poliovirus assembly sufficiently to act as an attenuation determinant [45]. In addition, the latter study showed that known ts producing mutations located in the 3D pol-coding region and 3 0 -UTR of poliovirus Sabin-1 [46,47] could be transferred to EV71 producing similar ts phenotypes. This is a clear indication that single mutations may be sufficient to change growth and neurovirulence phenotypes in wild-type EV71 isolates.…”

Generation and Characterization of Temperature Resistant Mutant of Recombinant PJ156/CAV-17 Virus

“…Identification of Amino Acids Determined Temperature Sensitivity. Previous studies revealed that two amino acids in 3D region (His-73 and Ile-362) were determinants of temperature sensitivity (Bouchard et al 1995;Georgescu et al 1995). A sequence analysis of 3D region of PJ156/CAV-17 showed that amino acid in 3D-73 and -362 were Tyr and Ile, respectively, similar with the parental CAV-17 sequence.…”

“…Previous studies revealed that mutations in the 5'NCR, base 2438 of VP3, bases 2741, and 2795 of VP1, base 6203 of 3Dpol, and base 7441 of the 3'NCR were associated with phenotypic reversion of the temperature sensitivity of PV, particularly Sabin 1 strain (Bouchard et al 1995). In addition, two amino acids in 3D region (His-73 and Ile-362) were determinants for temperature sensitivity of PV Sabin 1 (Georgescu et al 1995). Although it is reported that a chimeric plasmid from cDNA clones of poliovirus and coxsackievirus produces a recombinant virus that is temperature-sensitive (Semler et al 1986), it is not known whether those amino acids affect the temperature sensitivity of CAV, including CAV-17.…”

“…These suggest the correlation between temperature sensitivity and attenuation of the virus. From the study of PV Sabin 1 strain, it is suggested that the temperature sensitive effect associated with 3'-terminal part of the Sabin 1 genome are results from the cumulative and/or synergistic effects of at least three genetic determinants, i.e., the His-73 and Ile-362 codons of 3D, which coding RNA polymerase, and nucleotide G-7441 of 3'-untranslated region (Bouchard et al 1995;Georgescu et al 1995). Since the 3'-terminal part of the PJ156/CAV-17 was originated from CAV-17, the temperature sensitivity of this recombinant virus is supposed to be due to the sequence in 3'-terminal part of CAV-17.…”