Foreign glycoproteins expressed from recombinant vesicular stomatitis viruses are incorporated efficiently into virus particles.
In a previous study we demonstrated that vesicular stomatitis virus (VSV) can be used as a vector to express a soluble protein in mammalian cells. Here we have generated VSV recombinants that express four different membrane proteins: the cellular CD4 protein, a CD4-G hybrid protein containing the ectodomain of CD4 and the transmembrane and cytoplasmic tail of the VSV glycoprotein (G), the measles virus hemagglutinin, or the measles virus fusion protein. The proteins were expressed at levels ranging from 23-62% that of VSV G protein and all were transported to the cell surface. In addition we found that all four proteins were incorporated into the membrane envelope of VSV along with the VSV G protein. The levels of incorporation of these proteins varied from 6-31% of that observed for VSV G. These results suggest that many different membrane proteins may be co-incorporated quite efficiently with VSV G protein into budding VSV virus particles and that specific signals are not required for this co-incorporation process. In fact, the CD4-G protein was incorporated with the same efficiency as wild type CD4. Electron microscopy of virions containing CD4 revealed that the CD4 molecules were dispersed throughout the virion envelope among the trimeric viral spike glycoproteins. The recombinant VSV-CD4 virus particles were about 18% longer than wild type virions, reflecting the additional length of the helical nucleocapsid containing the extra gene. Recombinant VSVs carrying foreign antigens on the surface of the virus particle may be useful for viral targeting, membrane protein purification, and for generation of immune responses.
Schnell
M J
MJ
Department of Pathology, Yale University School of Medicine, New Haven, CT 06510, USA.
Buonocore
L
L
Kretzschmar
E
E
Johnson
E
E
Rose
J K
JK
D000818:Animals; D015703:Antigens, CD / Q000096:biosynthesis; D015704:CD4 Antigens / Q000096:biosynthesis; D002460:Cell Line; D006224:Cricetinae; D005822:Genetic Vectors; D006023:Glycoproteins / Q000096:biosynthesis*; D006389:Hemagglutinins, Viral / Q000096:biosynthesis; D006801:Humans; D007668:Kidney; D008459:Measles virus / Q000235:genetics / Q000378:metabolism; D008562:Membrane Glycoproteins; D016253:Microscopy, Immunoelectron; D011994:Recombinant Proteins / Q000096:biosynthesis; D014162:Transfection / Q000379:methods*; D014721:Vesicular stomatitis Indiana virus / Q000235:genetics* / Q000378:metabolism; D014759:Viral Envelope Proteins / Q000096:biosynthesis*; D014760:Viral Fusion Proteins / Q000096:biosynthesis
D015703:Antigens, CD; D015704:CD4 Antigens; C028391:G protein, vesicular stomatitis virus; D006023:Glycoproteins; D006389:Hemagglutinins, Viral; D008562:Membrane Glycoproteins; D011994:Recombinant Proteins; D014759:Viral Envelope Proteins; D014760:Viral Fusion Proteins