Sindbis-Group Alphavirus Replication in Periosteum and Endosteum of Long Bones in Adult Mice

Heise, Mark T.; Simpson, Dennis A.; Johnston, Robert E.

doi:10.1128/jvi.74.19.9294-9299.2000

Cited by 55 publications

(48 citation statements)

References 26 publications

(29 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Alphavirus replicons derived from SIN, SFV, and Venezuelan equine encephalitis virus have been successfully used to express a variety of foreign genes at high efficiency for various purposes such as vaccine development (9), reporter gene expression (19), and packaging of heterologous viral RNAs (22). To express WNV structural proteins, the WNV structural coding region encoding C, prM, and E was placed under transcriptional control of the subgenomic 26S promoter of an SR containing a mutation in the nsP2 gene, rendering it noncytopathic (726P3G mutation) (12).…”

Section: Resultsmentioning

confidence: 99%

trans -Packaged West Nile Virus-Like Particles: Infectious Properties In Vitro and in Infected Mosquito Vectors

Scholle¹,

Girard²,

Zhao³

et al. 2004

J Virol

119

View full text Add to dashboard Cite

show abstract

Section: Resultsmentioning

confidence: 99%

trans -Packaged West Nile Virus-Like Particles: Infectious Properties In Vitro and in Infected Mosquito Vectors

Scholle¹,

Girard²,

Zhao³

et al. 2004

J Virol

119

View full text Add to dashboard Cite

show abstract

“…First, 2A mediates cleavage at its own C terminus and therefore can remove itself and all upstream residues from polyproteins in which it is incorporated in a site-specific manner. This property of 2A has been exploited previously to construct recombinant picornavirus-based (20,24,44) and influenza A virus-based (23) expression vectors and alphavirus-based (14,39) and Kunjin virus-based (43) replicons. Here we used the 2A protease to construct live Sindbis virus expression vectors that express GFP/2A and VP7/2A fusion proteins as cleavable components of the viral structural polyprotein.…”

Section: Discussionmentioning

confidence: 99%

Sindbis Virus Vectors Designed To Express a Foreign Protein as a Cleavable Component of the Viral Structural Polyprotein

Thomas

Klimstra

Ryman

et al. 2003

J Virol

View full text Add to dashboard Cite

Alphavirus-based expression vectors commonly use a duplicated 26S promoter to drive expression of a foreign gene. Here we describe an expression strategy in which the foreign sequences are linked to the gene encoding the 2A protease of foot-and-mouth disease virus and then inserted in frame between the capsid and E3 genes of Sindbis virus. During replication, the 2A fusion protein is synthesized as a component of the viral structural polyprotein that is then released by intramolecular cleavages mediated by the capsid and 2A proteases. Recombinant Sindbis viruses that expressed fusion proteins composed of 2A linked to the green fluorescent protein (GFP) and to the VP7 protein of bluetongue virus were constructed. Viruses engineered to express GFP and VP7 from a duplicate 26S promoter were also constructed. All four viruses expressed the transgene and grew to similar titers in cultured cells. However, the GFP/2A-and VP7/2A-expressing viruses displayed greater expression stability and were less attenuated in newborn mice than the cognate doublesubgenomic promoter-based viruses. By combining the two expression strategies, we constructed bivalent viruses that incorporated and expressed both transgenes. The bivalent viruses grew to lower titers in cultured cells and were essentially avirulent in newborn mice. Groups of mice were vaccinated with each VP7-and VP7/2A-expressing virus, and antibody responses to native VP7 were measured in an indirect enzyme-linked immunosorbent assay. Despite their genetic and phenotypic differences, all viruses induced similarly high titers of VP7-specific antibodies. These results demonstrate that 2A fusion protein-expressing alphaviruses may be particularly well suited for applications that require enduring expression of a single protein or coexpression of two alternative proteins.Alphaviruses are enveloped, single-stranded, positive-sense RNA viruses that belong to the Togaviridae virus family (Alphavirus genus). Upon entry into the host cell, the viral genome is translated into the four nonstructural proteins (nsp1 to -4) that comprise the viral transcriptase-replicase complex. The viral replicase uses the genomic RNA as a template to synthesize a complementary, full-length, negative-sense RNA. The negative-sense RNA in turn serves as a template for the synthesis of two different positive-sense RNAs. Positive-strand synthesis initiating at the 3Ј end of the negative-strand RNA results in the production of full-length genomic RNA. Positivestrand synthesis can also initiate at an internal promoter sequence (26S or subgenomic promoter) to produce a subgenomic mRNA that is colinear with approximately the 3Ј-terminal one-third of the genomic RNA. Subgenomic mRNAs are translated into the viral structural proteins (40).

show abstract

“…For the Girdwood replicon system, the non-structural coding region of the genome from nucleotides 41-6463 was transferred from the full-length infectious clone pG100 [23] into the AR86-based replicon pREP89 [22], using restriction enzymes MfeI and BstBI. All non-structural coding differences between AR86 and GirdwoodS.A.…”

Section: Replicon Productionmentioning

confidence: 99%

An alphavirus replicon-derived candidate vaccine against Rift Valley fever virus

Heise

Whitmore²,

Thompson

et al. 2009

Epidemiol. Infect.

View full text Add to dashboard Cite

SUMMARY Rift Valley fever virus (RVFV) is a mosquito-transmitted bunyavirus (genus Phlebovirus)associated with severe disease in livestock and fatal encephalitis or haemorrhagic fever in a proportion of infected humans. Although live attenuated and inactivated vaccines have been used in livestock, and on a limited scale in humans, there is a need for improved anti-RVFV vaccines. Towards this goal, Sindbis virus replicon vectors expressing the RVFV Gn and Gc glycoproteins, as well as the non-structural nsM protein, were constructed and evaluated for their ability to induce protective immune responses against RVFV. These replicon vectors were shown to produce the RVFV glycoproteins to high levels in vitro and to induce systemic anti-RVFV antibody responses in immunized mice, as determined by RVFV-specific ELISA, fluorescent antibody tests, and demonstration of a neutralizing antibody response. Replicon vaccination also provided 100% protection against lethal RVFV challenge by either the intraperitoneal or intranasal route. Furthermore, preliminary results indicate that the replicon vectors elicit RVFV-specific neutralizing antibody responses in vaccinated sheep. These results suggest that alphavirus-based replicon vectors can induce protective immunity against RVFV, and that this approach merits further investigation into its potential utility as a RVFV vaccine.

show abstract

Sindbis-Group Alphavirus Replication in Periosteum and Endosteum of Long Bones in Adult Mice

Cited by 55 publications

References 26 publications

trans -Packaged West Nile Virus-Like Particles: Infectious Properties In Vitro and in Infected Mosquito Vectors

trans -Packaged West Nile Virus-Like Particles: Infectious Properties In Vitro and in Infected Mosquito Vectors

Sindbis Virus Vectors Designed To Express a Foreign Protein as a Cleavable Component of the Viral Structural Polyprotein

An alphavirus replicon-derived candidate vaccine against Rift Valley fever virus

Contact Info

Product

Resources

About