Attenuation of virulence of flaviviruses following passage in HeLa cells

Dunster, L. M.; Gibson, C.A.; Stephenson, J. R.; Minor, Philip D.; Barrett, Alan D.T.

doi:10.1099/0022-1317-71-3-601

Cited by 16 publications

(9 citation statements)

References 13 publications

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“…Comparison between these virus strains did not resolve differences in serological reactions with both immune serum and monoclonal antibodies, in surface charge measured by calcium phosphate column chromatography (data not shown), and in the oligonucleotide fingerprints of their RNA genome. Also, they grew to comparable titers at sub-and supraoptimal temperatures (32 °C and 39 °C, respectively) in baby hamster kidney (BHK) cell monolayers (data not shown), indicating that WN25 strain was not a temperature-sensitive (ts) mutant, as was reported for another WN strain [11]. In addition to IP infectivity, wild type WNV and WN25 differed in the mobility of their E protein, probably due to a difference in glycan content, and in the higher infectivity of WNV towards cultures of mouse peritoneal macrophages [17].…”

Section: Discussionsupporting

confidence: 50%

Loss of active neuroinvasiveness in attenuated strains of West Nile virus: pathogenicity in immunocompetent and SCID mice

Halevy

Akov

Ben‐Nathan

et al. 1994

Archives of Virology

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The neuropathogenicity of West Nile virus (WNV) and two derived attenuated strains WN25 and WN25A, was studied in young adult ICR mice and in severe combined immunodeficient (SCID) mice. Similarity in serology and RNA fingerprints were found between WNV and WN25. The viral envelope proteins of the attenuates differed from WNV in their slower mobility in SDS-PAGE due probably to the presence of N-linked glycan. The three strains were lethal to ICR mice by intracerebral (IC) inoculation, but when inoculated intraperitoneally (IP), WNV caused viremia, invaded the CNS and was lethal, whereas the attenuates showed no viremia or invasion of the CNS. The attenuates elicited antibodies to comparable levels as WNV in IP-infected mice, conferring upon them immunity to IC challenge with the wild type. In IP-inoculated SCID mice the three strains exhibited similar high viremiae that lasted until death of the animals. All strains invaded the CNS and proliferated in the mouse brain to similar high titers, but differed largely in the time of invasion: WNV invaded the CNS of SCID mice (and two other mouse strains) much earlier than the attenuates, which showed large intervals in their time of invasion into individual mouse brains within the group. The data presented for SCID mice indicate that WN25 and WN25A have truly lost the neuroinvasive property, and that this property materialized by a prescribed, active process specific for WNV.

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Section: Discussionsupporting

confidence: 50%

Loss of active neuroinvasiveness in attenuated strains of West Nile virus: pathogenicity in immunocompetent and SCID mice

Halevy

Akov

Ben‐Nathan

et al. 1994

Archives of Virology

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“…Studies demonstrating attenuation of flavivirus neurovirulence have also been reported, but this appears to be a less frequent response to passaging. Dunster et al (1990) demonstrated attenuation of WN virus neurovirulence after six passages in HeLa cells; the passaged virus showed altered reactivity to an E protein MAb compared to parental virus, suggesting that attenuation was due to mutation in the E protein.…”

Section: Discussionmentioning

confidence: 99%

Neurovirulence and neuroinvasiveness of Murray Valley encephalitis virus mutants selected by passage in a monkey kidney cell line

McMinn

Marshall

Dalgarno

1995

Journal of General Virology

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Variants of the prototype Murray Valley encephalitisvirus (MVE-1-51) were selected by serial plaque purification and amplification in monkey kidney (Vero) cells.Four clones (C1-C4) at passage levels two and nine (P2 and P9) were examined in 21-day-old Swiss outbred mice for neuroinvasiveness ( Genomic RNA of the cloned virus stocks was sequenced through the structural protein genes (E, prM/M and C) and the 5' untranslated region. Clone C2P2 was of high neuroinvasiveness whereas C2P9 was of low neuroinvasiveness; there were also decreased yields of C2P9 in C6/36 cells compared to C2P2 and MVE-1-51. These changes were associated with the substitution of valine for phenylalanine at amino acid position 141 of the C2P9 E protein. Clone C4P2 was of high neurovirulence and low neuroinvasiveness; C4P9 was of low neurovirulence, a change accompanied by a further reduction in neuroinvasiveness. Concomitantly, C4P9 showed a pronounced reduction in growth rates and yields in 21-day-old Swiss mouse brain, in mouse neuroblastoma cells and in C6/36 cells compared to parental virus. The phenotypic changes in clone 4 appear to be due to mutation(s) within non-structural protein genes.

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“…Such analyses should be on low passage virus, since laboratory adaptation of dengue virus and other flaviviruses, in cell culture and in suckling mice, can alter the antigenic, biologic and genetic properties of the virus [5,15,16,18,19,24].…”

Section: Introductionmentioning

confidence: 99%

Genetic and biological differentiation of dengue 3 isolates obtained from clinical cases in Java, Indonesia, 1976?1978

Lee

Gubler

Weir

et al. 1993

Archives of Virology

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Previous epidemiological, virological and clinical studies have documented a series of outbreaks of dengue fever and dengue haemorrhagic fever/dengue shock syndrome which occurred in Java, Indonesia in 1976-1978. In the current study we compare growth characteristics in cell culture, and nucleotide sequence data for the viral prM and E genes, of five low passage DEN-3 isolates obtained during these epidemics from clinically defined cases. All isolates had the same passage history: human sera were passed twice in mosquitoes and three times in a mosquito cell line (Aedes albopictus, C 6/36 cells). Growth differences were observed between individual isolates in Vero cells; growth differences were not observed in C 6/36 cells. Nucleotide sequencing of the prM and E gene region indicated that no two isolates were identical (sequence divergence ranged from 0.4 to 1.6% in pairwise comparisons) but that they were closely enough related to present a single genetic type. There were one or two differences in deduced amino acid sequence in E between isolates. Differences were at residues 65, 187, 298 or 443. One isolate differed from all others at residue 16 in the M protein. No relationship was apparent between the amino acid sequence of M or E and the nature of the disease profile, the year of isolation or the geographic region of isolation. The isolates showed 3.5 to 4.4% nucleotide sequence divergence from the highly-adapted H 87 prototype, isolated in the Philippines in 1956. The isolates showed a total of twelve common amino acid differences in prM and E proteins from H 87. Ten of these twelve residues were at positions which differed between the four dengue serotypes. Two differences (at residues 37 in M and 293 in E) were at positions which are conserved in sequence between the four dengue serotypes. The data are discussed in relation to the dengue outbreaks in Java in the period 1976-1978.

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Attenuation of virulence of flaviviruses following passage in HeLa cells

Cited by 16 publications

References 13 publications

Loss of active neuroinvasiveness in attenuated strains of West Nile virus: pathogenicity in immunocompetent and SCID mice

Loss of active neuroinvasiveness in attenuated strains of West Nile virus: pathogenicity in immunocompetent and SCID mice

Neurovirulence and neuroinvasiveness of Murray Valley encephalitis virus mutants selected by passage in a monkey kidney cell line

Genetic and biological differentiation of dengue 3 isolates obtained from clinical cases in Java, Indonesia, 1976?1978

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