Modified H5 promoter improves stability of insert genes while maintaining immunogenicity during extended passage of genetically engineered MVA vaccines

Abstract We have engineered recombinant (r) Modified Vaccinia Ankara (MVA) to express multiple antigens under the control of either of two related vaccinia synthetic promoters (pSyn) with early and late transcriptional activity or the modified H5 (mH5) promoter which has predominant early activity....

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Bibliographic Details
Published in:Vaccine 2010-02, Vol.28 (6), p.1547-1557
Main Authors: Wang, Zhongde, Martinez, Joy, Zhou, Wendi, La Rosa, Corinna, Srivastava, Tumul, Dasgupta, Anindya, Rawal, Ravindra, Li, Zhongqui, Britt, William J, Diamond, Don
Format: Article
Language:English
Subjects:
Allergy and Immunology
Animals
Applied microbiology
Biological and medical sciences
Cancer
Cellular immunity
Cloning
Deoxyribonucleic acid
DNA
DNA, Viral - chemistry
DNA, Viral - genetics
Fundamental and applied biological sciences. Psychology
Gene expression
Genetic engineering
Genetic Vectors
Genomic Instability
Herpesviridae - genetics
Herpesvirus
Humans
Immunogenicity
Infections
Infectious diseases
Kinases
Laboratories
Mice
Microbiology
Miscellaneous
Modified Vaccinia Ankara
Molecular Sequence Data
Plasmids
Polymerase Chain Reaction
Poxvirus
Promoter Regions, Genetic
Protein expression
Proteins
Sequence Analysis, DNA
Serial Passage
Vaccine
Vaccines
Vaccines - genetics
Vaccines - immunology
Vaccines, antisera, therapeutical immunoglobulins and monoclonal antibodies (general aspects)
Vaccines, Synthetic - genetics
Vaccines, Synthetic - immunology
Vaccinia virus - genetics
Virology
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Summary:Abstract We have engineered recombinant (r) Modified Vaccinia Ankara (MVA) to express multiple antigens under the control of either of two related vaccinia synthetic promoters (pSyn) with early and late transcriptional activity or the modified H5 (mH5) promoter which has predominant early activity. We sequentially passaged these constructs and analyzed their genetic stability by qPCR, and concluded that rMVA expressing multiple antigens using the mH5 promoter exhibit remarkable genetic stability and maintain potent immunogenicity after serial passage. In contrast, rMVA expressing antigens using engineered vaccinia synthetic E/L (pSyn I or II) promoters are genetically unstable. Progressive accumulation of antigen loss variants resulted in a viral preparation with lower immunogenicity after serial passage. Metabolic labeling, followed by cold chase revealed little difference in stability of proteins expressed from mH5 or pSyn promoter constructs. We conclude that maintenance of genetic stability which is achieved using mH5, though not with pSyn promoters, is linked to timing, not the magnitude of expression levels of foreign antigen, which is more closely associated with immunogenicity of the vaccine.
ISSN:0264-410X
1873-2518
DOI:10.1016/j.vaccine.2009.11.056