Synthesis of the allergen ovomucoid by a replicating Mengo virus.
Interferons induced by viral infections can have powerful immuno- modulatory effects, and several epidemiologic studies have found an association between certain viral infections and reduced prevalence of allergy. We hypothesized that allergenic proteins could be synthesized by a replicating virus, and this construct could be useful as an immunomodulator. To test this hypothesis, we cloned an allergenic protein (ovomucoid [OVM]) into a murine picornavirus (Mengo virus) vector. This plasmid has a multicloning site surrounded by auto-catalytic sequences so that a foreign protein will be cleaved from viral proteins during replication. OVM sequences were cloned in the context of full-length viral genome cDNA, T7 RNA transcripts of this plasmid were transfected into HeLa cells, and recombinant virus plaques appeared on the second passage. Sequence analysis of recombinant viruses derived from individual plaques demonstrated that three viral isolates contained up to 2/3 of the OVM coding sequence, which was retained by the viruses after 5 additional passages in HeLa cells. The experiments verify the stable expression of immunoreactive OVM subunits by replicating viruses. These virus/allergen constructs could provide a tool to evaluate whether intracellular presentation of allergenic proteins in the context of a viral infection could prevent allergic sensitization upon re-challenge.
Amineva
S P
SP
Department of Pediatrics, University of Wisconsin-Madison, Madison, WI, USA. spamineva@wisc.edu
Mosser
A G
AG
Binder
J J
JJ
Aminev
A G
AG
Palmenberg
A C
AC
Gern
J E
JE
D000485:Allergens / Q000096:biosynthesis* / Q000235:genetics; D000818:Animals; D018188:Cardiovirus Infections / Q000821:virology; D005822:Genetic Vectors / Q000378:metabolism / Q000502:physiology; D016679:Genome, Viral / Q000235:genetics; D006367:HeLa Cells / Q000378:metabolism; D006801:Humans; D008574:Mengovirus / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D051379:Mice; D010059:Ovomucin / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology; D010957:Plasmids / Q000235:genetics; D016865:Reassortant Viruses / Q000378:metabolism / Q000502:physiology*; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000276:immunology; D011995:Recombination, Genetic; D012692:Serial Passage; D013045:Species Specificity; D014162:Transfection; D014779:Virus Replication
D000485:Allergens; D011994:Recombinant Proteins; D010059:Ovomucin