Isolation of Recombinant MVA Using F13L Selection

Sánchez-Puig, Juana M.; Lorenzo, Marı́a M.; Blasco, Rafael

doi:10.1007/978-1-61779-876-4_5

Cited by 10 publications

(15 citation statements)

References 21 publications

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“…In this sense, it has been described that genome location and TK function can contribute to the relative immunogenicity of antigens when expressed from rMVA 45 . In addition, in previous rMVA-GnGc vaccine construct 10 , the heterologous gene was cloned under the control of the vaccinia 7.5 k early/late promoter, while in the MVA-GnGc-NS1 describe here the RVFV glycoproteins genes were placed under the control of an optimized strong early/ late promoter 46,47 . The different locus and promoters where GnGc was located in the MVA could explain the differences between rMVA-GnGc and MVA-GnGc-NS1 observed in protection against RVFV.…”

Section: Discussionmentioning

confidence: 99%

A protective bivalent vaccine against Rift Valley fever and bluetongue

et al. 2020

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Rift Valley fever (RVF) and bluetongue (BT) are two important ruminant diseases transmitted by arthropods. Both viruses have shown important geographic spread leading to endemicity of BT virus (BTV) in Africa and Europe. In this work, we report a dual vaccine that simultaneously induces protective immune responses against BTV and RVFV based on modified vaccinia Ankara virus (MVA) expressing BTV proteins VP2, NS1, or a truncated form of NS1 (NS1-Nt), and RVFV Gn and Gc glycoproteins. IFNAR (−/−) mice immunized with two doses of MVA-GnGc-VP2 developed a significant neutralizing antibody response against BTV-4 and RVFV. Furthermore, the homologous prime-boost immunization with MVA-GnGc-NS1 or MVA-GnGc-NS1-Nt triggered neutralizing antibodies against RVFV and NS1-specific cytotoxic CD8+ T cells in mice. Moreover, all mice immunized with MVA-GnGc-NS1 or MVA-GnGc-NS1-Nt remained healthy after lethal challenge with RVFV or BTV-4. The homologous prime-boost vaccination with MVA-GnGc-NS1, which was the best immunization strategy observed in mice, was assayed in sheep. Clinical signs and viremia were absent or highly reduced in vaccinated sheep after challenge with BTV-4 or RVFV. These results indicate that MVA-GnGc-NS1 vaccination elicits immune protection against RVFV and BTV in sheep.

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Section: Discussionmentioning

confidence: 99%

A protective bivalent vaccine against Rift Valley fever and bluetongue

et al. 2020

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“…Poxvirus recombinants expressing BFP were obtained by transfecting plasmid pRB-LF-TagBFP (containing a TagBFP cassette cloned downstream of the F13L gene in pRB21 plasmid, MML, manuscript in preparation) into cells infected with VACV vRB12 [18], MVAΔF13L [19, 20], vRB10 [21], CPXV Brighton red, or CPXV EP4 to isolate WR-BFP, MVA-BFP, IHDJ-BFP, BR-BFP and EP4-BFP, respectively. Virus isolation was accomplished by plaque identification aided by BFP fluorescence and plaque purification.…”

Section: Methodsmentioning

confidence: 99%

Vaccinia virus and Cowpox virus are not susceptible to the interferon-induced antiviral protein MxA

2017

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MxA protein is expressed in response to type I and type III Interferon and constitute an important antiviral factor with broad antiviral activity to diverse RNA viruses. In addition, some studies expand the range of MxA antiviral activity to include particular DNA viruses like Monkeypox virus (MPXV) and African Swine Fever virus (ASFV). However, a broad profile of activity of MxA to large DNA viruses has not been established to date. Here, we investigated if some well characterized DNA viruses belonging to the Poxviridae family are sensitive to human MxA. A cell line inducibly expressing MxA to inhibitory levels showed no anti-Vaccinia virus (VACV) virus activity, indicating either lack of susceptibility of the virus, or the existence of viral factors capable of counteracting MxA inhibition. To determine if VACV resistance to MxA was due to a virus-encoded anti-MxA activity, we performed coinfections of VACV and the MxA-sensitive Vesicular Stomatitis virus (VSV), and show that VACV does not protect VSV from MxA inhibition in trans. Those results were extended to several VACV strains and two CPXV strains, thus confirming that those Orthopoxviruses do not block MxA action. Overall, these results point to a lack of susceptibility of the Poxviridae to MxA antiviral activity.

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“…Virus infections were carried out in medium containing 2% FBS. Plaque assays and crystal violet staining was carried out as described [30] [31].…”

Section: Methodsmentioning

confidence: 99%

A Vaccinia Virus Recombinant Transcribing an Alphavirus Replicon and Expressing Alphavirus Structural Proteins Leads to Packaging of Alphavirus Infectious Single Cycle Particles

2013

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Poxviruses and Alphaviruses constitute two promising viral vectors that have been used extensively as expression systems, or as vehicles for vaccine purposes. Poxviruses, like vaccinia virus (VV) are well-established vaccine vectors having large insertion capacity, excellent stability, and ease of administration. In turn, replicons derived from Alphaviruses like Semliki Forest virus (SFV) are potent protein expression and immunization vectors but stocks are difficult to produce and maintain. In an attempt to demonstrate the use of a Poxvirus as a means for the delivery of small vaccine vectors, we have constructed and characterized VV/SFV hybrid vectors. A SFV replicon cDNA was inserted in the VV genome and placed under the control of a VV early promoter. The replicon, transcribed from the VV genome as an early transcript, was functional, and thus capable of initiating its own replication and transcription. Further, we constructed a VV recombinant additionally expressing the SFV structural proteins under the control of a vaccinia synthetic early/late promoter. Infection with this recombinant produced concurrent transcription of the replicon and expression of SFV structural proteins, and led to the generation of replicon-containing SFV particles that were released to the medium and were able to infect additional cells. This combined VV/SFV system in a single virus allows the use of VV as a SFV delivery vehicle in vivo. The combination of two vectors, and the possibility of generating in vivo single-cycle, replicon containing alphavirus particles, may open new strategies in vaccine development or in the design of oncolytic viruses.

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Isolation of Recombinant MVA Using F13L Selection

Cited by 10 publications

References 21 publications

A protective bivalent vaccine against Rift Valley fever and bluetongue

A protective bivalent vaccine against Rift Valley fever and bluetongue

Vaccinia virus and Cowpox virus are not susceptible to the interferon-induced antiviral protein MxA

A Vaccinia Virus Recombinant Transcribing an Alphavirus Replicon and Expressing Alphavirus Structural Proteins Leads to Packaging of Alphavirus Infectious Single Cycle Particles

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