scholarly journals Transient Host Range Selection for Genetic Engineering of Modified Vaccinia Virus Ankara

BioTechniques ◽  
2000 ◽  
Vol 28 (6) ◽  
pp. 1137-1148 ◽  
Author(s):  
C. Staib ◽  
I. Drexler ◽  
M. Ohlmann ◽  
S. Wintersperger ◽  
V. Erfle ◽  
...  
Keyword(s):  
Genetic Engineering ◽  
Vaccinia Virus ◽  
Host Range ◽  
Modified Vaccinia Virus Ankara ◽  
Selection For ◽  
Range Selection

scholarly journals Improved Host Range Selection for Recombinant Modified Vaccinia Virus Ankara

BioTechniques ◽  
2003 ◽  
Vol 34 (4) ◽  
pp. 694-700 ◽  
Author(s):  
Caroline Staib ◽  
Marianne Löwel ◽  
Volker Erfle ◽  
Gerd Sutter
Keyword(s):  
Vaccinia Virus ◽  
Host Range ◽  
Modified Vaccinia Virus Ankara ◽  
Selection For ◽  
Range Selection

scholarly journals Notes on Transient Host Range Selection for Engineering Vaccinia Virus Strain MVA

BioTechniques ◽  
2002 ◽  
Vol 33 (1) ◽  
pp. 186-188 ◽  
Author(s):  
David C. Tscharke ◽  
Geoffrey L. Smith
Keyword(s):  
Vaccinia Virus ◽  
Host Range ◽  
Virus Strain ◽  
Selection For ◽  
Vaccinia Virus Strain ◽  
Range Selection

Marker gene swapping facilitates recombinant Modified Vaccinia Virus Ankara production by host-range selection

2009 ◽  
Vol 156 (1-2) ◽  
pp. 37-43 ◽  
Author(s):  
Giulia Di Lullo ◽  
Elisa Soprana ◽  
Maddalena Panigada ◽  
Alessio Palini ◽  
Volker Erfle ◽  
...  
Keyword(s):  
Vaccinia Virus ◽  
Host Range ◽  
Marker Gene ◽  
Modified Vaccinia Virus Ankara ◽  
Range Selection

scholarly journals Repair of a previously uncharacterized second host-range gene contributes to full replication of modified vaccinia virus Ankara (MVA) in human cells

2020 ◽  
Vol 117 (7) ◽  
pp. 3759-3767 ◽  
Author(s):  
Chen Peng ◽  
Bernard Moss
Keyword(s):  
Vaccinia Virus ◽  
Host Range ◽  
Cell Lines ◽  
Chicken Embryo ◽  
A549 Cells ◽  
Human Cells ◽  
Modified Vaccinia Virus Ankara ◽  
Core Proteins ◽  
Embryo Fibroblasts ◽  
Chicken Embryo Fibroblasts

Modified vaccinia virus Ankara (MVA), a widely used vaccine vector for expression of genes of unrelated pathogens, is safe, immunogenic, and can incorporate large amounts of added DNA. MVA was derived by extensively passaging the chorioallantois vaccinia virus Ankara (CVA) vaccine strain in chicken embryo fibroblasts during which numerous mutations and deletions occurred with loss of replicative ability in most mammalian cells. Restoration of the deleted C12L gene, encoding serine protease inhibitor 1, enhances replication of MVA in human MRC-5 cells but only slightly in other human cells. Here we show that repair of the inactivated C16L/B22R gene of MVA enhances replication in numerous human cell lines. This previously uncharacterized gene is present at both ends of the genome of many orthopoxviruses and is highly conserved in most, including smallpox and monkeypox viruses. The C16L/B22R gene is expressed early in infection from the second methionine of the previously annotated Copenhagen strain open reading frame (ORF) as a 17.4-kDa protein. The C16/B22 and C12 proteins together promote MVA replication in human cells to levels that are comparable to titers in chicken embryo fibroblasts. Both proteins enhance virion assembly, but C16/B22 increases proteolytic processing of core proteins in A549 cells consistent with higher infectious virus titers. Furthermore, human A549 cells expressing codon-optimized C16L/B22R and C12L genes support higher levels of MVA replication than cell lines expressing neither or either alone. Identification of the genes responsible for the host-range defect of MVA may allow more rational engineering of vaccines for efficacy, safety, and propagation.

scholarly journals Marker Rescue of the Host Range Restriction Defects of Modified Vaccinia Virus Ankara

Virology ◽  
1998 ◽  
Vol 251 (2) ◽  
pp. 334-342 ◽  
Author(s):  
Linda S. Wyatt ◽  
Miles W. Carroll ◽  
Claus-Peter Czerny ◽  
Michael Merchlinsky ◽  
Jerry R. Sisler ◽  
...  
Keyword(s):  
Vaccinia Virus ◽  
Host Range ◽  
Range Restriction ◽  
Marker Rescue ◽  
Modified Vaccinia Virus Ankara ◽  
Host Range Restriction

scholarly journals Genomic sequence of chorioallantois vaccinia virus Ankara, the ancestor of modified vaccinia virus Ankara

2007 ◽  
Vol 88 (12) ◽  
pp. 3249-3259 ◽  
Author(s):  
Christine Meisinger-Henschel ◽  
Michaela Schmidt ◽  
Susanne Lukassen ◽  
Burkhard Linke ◽  
Lutz Krause ◽  
...  
Keyword(s):  
Vaccinia Virus ◽  
Host Range ◽  
Genomic Sequence ◽  
Vaccine Virus ◽  
Open Reading Frames ◽  
Detailed Knowledge ◽  
Synonymous Mutations ◽  
Modified Vaccinia Virus Ankara ◽  
Vacv Strain ◽  
Chicken Embryo Fibroblasts

Chorioallantois vaccinia virus Ankara (CVA) is the parental virus of modified vaccinia virus Ankara (MVA), which was derived from CVA by more than 570 passages in chicken embryo fibroblasts (CEF). MVA became severely host-cell-restricted to avian cells and has strongly diminished virulence in mammalian hosts, while maintaining good immunogenicity. We determined the complete coding sequence of the parental CVA and mapped the exact positions of the six major deletions that emerged in the MVA genome. All six major deletions occurred in regions of the CVA genome where one or more truncated or fragmented open reading frames (ORFs) pre-existed. The CVA genome contains 229 ORFs of which 51 are fragments of full-length orthopoxvirus (OPV) genes, including fragmented orthologues of C9L and M1L (encoding two well-conserved ankyrin-like proteins), A39R (encoding a semaphorin-like protein) and A55R (encoding a kelch-like protein). Phylogenetic analysis demonstrated that MVA was most closely related to CVA, followed by the vaccinia virus (VACV) strain DUKE, a patient-derived isolate of the Dryvax vaccine virus. Loss or mutation of genes outside the six major deletions are assumed to contribute to the restricted host range phenotype of MVA. In support of this notion, deletions, insertions and non-synonymous mutations were found in 122 of the 195 ORFs remaining in MVA when compared with their CVA counterparts. Thus, detailed knowledge of the CVA genomic sequence is a prerequisite to further dissect the genetic basis of the MVA host range phenotype as well as the particular immunological properties of MVA.

scholarly journals Introduction of the Six Major Genomic Deletions of Modified Vaccinia Virus Ankara (MVA) into the Parental Vaccinia Virus Is Not Sufficient To Reproduce an MVA-Like Phenotype in Cell Culture and in Mice

2010 ◽  
Vol 84 (19) ◽  
pp. 9907-9919 ◽  
Author(s):  
Christine Meisinger-Henschel ◽  
Michaela Späth ◽  
Susanne Lukassen ◽  
Michael Wolferstätter ◽  
Heike Kachelriess ◽  
...  
Keyword(s):  
Cell Culture ◽  
Vaccinia Virus ◽  
Host Range ◽  
Cell Lines ◽  
Moderate Degree ◽  
Range Restriction ◽  
Genomic Deletions ◽  
Artificial Chromosomes ◽  
Modified Vaccinia Virus Ankara

ABSTRACT Modified vaccinia virus Ankara (MVA) has a highly restricted host range in cell culture and is apathogenic in vivo. MVA was derived from the parental chorioallantois vaccinia virus Ankara (CVA) by more than 570 passages in chicken embryo fibroblast (CEF) cells. During CEF cell passaging, six major deletions comprising 24,668 nucleotides occurred in the CVA genome. We have cloned both the MVA and the parental CVA genome as bacterial artificial chromosomes (BACs) and have sequentially introduced the six major MVA deletions into the cloned CVA genome. Reconstituted mutant CVA viruses containing up to six major MVA deletions showed no detectable replication restriction in 12 of 14 mammalian cell lines tested; the exceptions were rabbit cell lines RK13 and SIRC. In mice, CVA mutants with up to three deletions showed slightly enhanced virulence, suggesting that gene deletion in replicating vaccinia virus (VACV) can result in gain of fitness in vivo. CVA mutants containing five or all six deletions were still pathogenic, with a moderate degree of attenuation. Deletion V was mainly responsible for the attenuated phenotype of these mutants. In conclusion, loss or truncation of all 31 open reading frames in the six major deletions is not sufficient to reproduce the specific MVA phenotype of strong attenuation and highly restricted host range. Mutations in viral genes outside or in association with the six major deletions appear to contribute significantly to this phenotype. Host range restriction and avirulence of MVA are most likely a cooperative effect of gene deletions and mutations involving the major deletions.

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