High-level eucaryotic in vivo expression of biologically active measles virus hemagglutinin by using an adenovirus type 5 helper-free vector system.
The entire measles virus (MV) hemagglutinin (HA)-coding region was reconstructed from cloned cDNAs and used as part of a hybrid transcription unit to replace a region of the adenovirus type 5 genome corresponding to the entire E1a transcription unit and most of the E1b transcription unit. The resulting recombinant virus was stable and able to replicate to high titers in 293 cells (which constitutively express the complementary E1a-E1b functions) in the absence of helper virus. During infection of 293 cells, the hybrid virus expressed MV HA protein which was indistinguishable from that expressed in MV-infected cells in terms of immunoreactivity, gel mobility, glycosylation, subcellular localization, and biologic activity. Infection of 293 cells with the hybrid virus led to high-level synthesis of the MV HA protein (equivalent to 65 to 130% of the level seen in MV-infected cells). At late times after high-multiplicity hybrid virus infection of HeLa and Vero cells (which do not express E1 functions), the level of HA protein synthesis was at least 35% of that seen in 293 cells. This MV-adenovirus recombinant will be useful in the study of the biologic properties of the MV HA protein and in assessment of the potential usefulness of hybrid adenoviruses as live-virus vaccine vectors.
Alkhatib
G
G
Department of Microbiology and Immunology, McGill University, Montreal, Quebec, Canada.
Briedis
D J
DJ
D000256:Adenoviridae / Q000235:genetics; D000818:Animals; D001681:Biological Assay; D002451:Cell Compartmentation; D002462:Cell Membrane / Q000502:physiology; D002478:Cells, Cultured; D003001:Cloning, Molecular; D004274:DNA, Recombinant; D005455:Fluorescent Antibody Technique; D005822:Genetic Vectors; D006389:Hemagglutinins, Viral / Q000235:genetics*; D008459:Measles virus / Q000235:genetics*; D012333:RNA, Messenger / Q000235:genetics; D012367:RNA, Viral / Q000235:genetics
D004274:DNA, Recombinant; D006389:Hemagglutinins, Viral; D012333:RNA, Messenger; D012367:RNA, Viral