Assay of Defective-interfering Semliki Forest Virus by the Inhibition of Synthesis of Virus-specified RNAs

Barrett, Alan D.T.; Crouch, Colin; Dimmock, Nigel J.

doi:10.1099/0022-1317-54-2-273

Cited by 29 publications

(26 citation statements)

References 24 publications

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“…Barrett et al (1981) reported that interference by SF virus DI particles varies depending upon the cell line used for assay. Stark & Kennedy (1978) could not generate SF virus DI particles in HeLa cells, even after 200 serial passages.…”

Section: Discussionmentioning

confidence: 99%

“…Defective interfering (DI) particles are a class of virus deletion mutant generated during infection initiated at both high and low multiplicities (for reviews, see Perrault, 1981 ;Barrett et al, 1981). They have a number of common properties, which include the inability to propagate in the absence of homologous standard virus, and the ability to decrease the yield of standard virus and increase their proportion of the yield from cells coinfected with standard virus (Huang & Baltimore, 1970).…”

Section: Introductionmentioning

confidence: 99%

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In vitro homotypic and heterotypic interference by defective interfering particles of West Nile virus

Debnath

Tiernery

Sil

et al. 1991

Journal of General Virology

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Defective interfering (DI) particles of the flavivirus West Nile (WN) were generated after as few as two high multiplicity serial passages in Vero and LLC-MK2 cells. Six cell lines (Vero, LLC-MK2, L929, HeLa, BHK-21 and SW13) were used to assay interference by DI particles in a yield reduction assay. Interference was found to vary depending on the cell type used. The highest levels of interference were obtained in LLC-MK2 cells, whereas no detectable effect was observed in BHK-21 and SW13 cells. The ability of DI virus to be propagated varied depending on the cell line used; no detectable propagation of DI virus was observed in SW 13 cells. Optimum interference was obtained following co-infection of cells with DI virus and standard virus at a multiplicity of 5. Interference between DI and standard viruses occurred only when they were co-infected or when cells were infected with DI virus 1 h before standard virus. Investigation of heterotypic interference by DI particles of WN virus strains from Sarawak, India and Egypt revealed that interference was dependent on the strain of WN virus or flavivirus used as standard virus. A measure of the similarity between five strains of WN virus and other flaviviruses was made on the basis of interference by DI viruses, and was found to be similar to that based on haemagglutination inhibition tests using a panel of monoclonal antibodies.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

In vitro homotypic and heterotypic interference by defective interfering particles of West Nile virus

Debnath

Tiernery

Sil

et al. 1991

Journal of General Virology

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“…Since the standard virus genome represents about 6.3 ~o of the particle mass (Laine et al, 1973) and DI particles have smaller RNA species, this difference should be detected by estimates of particle density. However, the data are conflicting: Shenk & Stollar (1973) and have reported that DI virus particles are more dense than standard virus particles while others found no difference in their densities (Weiss & Schlesinger, 1973;Guild & Stollar, 1975;Logan, 1979 Detailed studies were made of a DI virus preparation which had been propagated for 13 undiluted passages (called DI SFV pl 3g) in BHK cells as described by Barrett et at. (1981).…”

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confidence: 99%

“…(1981). DI virus and standard virus (Barrett et al, 1981) were concentrated by centrifugation at 90 000 g for 3 h at 4 °C and resuspended in 50 mM-Tris-HC1, 100 mM-NaC1, 1 mM-EDTA, pH 7.4, but were not purified further. Virus was prepared for electron microscopy by placing I0 ~tl aliquots onto 2~ agarose in phosphate-buffered saline (PBS) in the wells of a microtitre plate.…”

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confidence: 99%

“…We have previously described a bioassay for measuring interference by DI SFV (the RNA synthesis inhibition assay; Barrett et al, 1981) from which, assuming that interference requires at least one particle per cell, a minimum estimate of the number of biologically active DI virus particles (DI units) can be obtained. This was checked against the number of small particles by determining their concentration by electron microscopy with reference to latex spheres of known concentration; there were approximately 130 small particles for each DI unit.…”

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Defective Interfering Particles of Semliki Forest Virus Are Smaller than Particles of Standard Virus

Barrett

Cubitt

Dimmock

1984

Journal of General Virology

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SUMMARYBy electron microscopy, particles of defective interfering Semliki Forest virus (DI SFV) had a mean diameter of 46-8 nm compared with 55.9 nm for standard virus particles, a decrease of 16~. The difference was confirmed by measurements of the two-dimensional projected areas of DI and standard virus particles. We examined nine different DI virus preparations produced by four to 13 undiluted passages in BHK cells and all were found to contain a majority of the smaller type of particle. Calculation of the absolute number of small particles showed that there were 130 particles per interfering unit measured by the inhibition of virus RNA synthesis. However, a more sensitive assay based on interference with virus protein synthesis gave a particle :interference ratio of 6.5.Defective interfering (DI) viruses are generated during serial undiluted passage of most, if not all, animal viruses (Huang & Baltimore, 1977) and have a genome which is a deleted form of the infectious (standard) virus genome. Since these viruses are defective and are unable to replicate unaided, propagation is achieved by co-infecting cells with DI and standard virus. The yield from such co-infected cells contains reduced quantities of standard virus, hence the term interference (Holland et al., 1980;Perrault, 1981). DI viruses have been described for the alphaviruses Sindbis and Semliki Forest (SFV) (Schlesinger et al., 1972;) and the genome has been shown to be smaller (often < 1 × 106) than that of standard virus (4-2 x 106) (Shenk & Stollar, 1972;Eaton & Faulkner, 1973; Weiss & Schlesinger, 1973;Eaton, 1975;Guild & Stollar, 1975;Logan, 1979). The RNA of DI SFV contains no open reading frame (Lehtovaara et al., , 1982 and does not synthesize polypeptides either in vivo or in vitro Logan, 1979; A. D. T. Barrett & N. J. Dimmock, unpublished data). Biochemical analysis shows that DI viruses contain the same proteins as standard virus Logan, 1979; A. D. T. Barrett & N. J. Dimmock, unpublished data); thus, it is assumed that DI SFV proteins are synthesized by the standard virus.Examination of the size of alphavirus DI particles has produced conflicting results. Weiss & Schlesinger (1973) and Logan (1979) reported that DI and standard viruses grown in BHK and mosquito cells respectively were the same size, while Johnston et al. (1975) showed that Sindbis DI virus particles were smaller (37 nm) than standard virus (50 nm) and DI virus preparations with high interfering activity had the greatest numbers of the small particles. These smaller particles were similar in size to morphological variants observed by Brown & Gliedman (1973) during Sindbis virus infection of mosquito cells; however, the latter workers did not determine whether or not these variants were DI particles. Since the standard virus genome represents about 6.3 ~o of the particle mass (Laine et al., 1973) and DI particles have smaller RNA species,

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Detection of viral ribonucleic acid and histologic analysis of inflamed synovium in Ross River virus infection

Soden

Vasudevan

Roberts

et al. 2000

Arthritis & Rheumatism

100

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Objective. To document the histology of Ross River virus (RRV) arthritis and to examine inflamed synovium for viral RNA. Methods. Biopsy tissue from the inflamed knees of 12 patients with RRV infection was studied using conventional and immunostaining techniques. Reverse transcriptase-polymerase chain reaction technology was used to probe for the presence of viral RNA in the synovial biopsy samples and in serum. Results. Hyperplasia of the synovial lining layer, vascular proliferation, and mononuclear cell infiltration were the main histologic changes. RRV RNA was found in knee biopsy tissue that was obtained from 2 patients at 5 weeks after the onset of symptoms. Conclusion. RRV RNA was identified in inflamed synovium more than a month after symptoms began. Inflammation was apparent in the absence of detectable virus in the majority of patients.

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Assay of Defective-interfering Semliki Forest Virus by the Inhibition of Synthesis of Virus-specified RNAs

Cited by 29 publications

References 24 publications

In vitro homotypic and heterotypic interference by defective interfering particles of West Nile virus

In vitro homotypic and heterotypic interference by defective interfering particles of West Nile virus

Defective Interfering Particles of Semliki Forest Virus Are Smaller than Particles of Standard Virus

Detection of viral ribonucleic acid and histologic analysis of inflamed synovium in Ross River virus infection

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