Association of glycosylphosphatidylinositol-anchored protein with retroviral particles.
We describe for the first time the association of glycosylphosphatidylinositol (GPI) -anchored proteins with retroviral and lentiviral particles, similar to a process well established for cells, termed "painting." The aim of the study was to assess the feasibility of modification of retroviral vectors by exogenous addition of recombinant protein, removing the need for genetic engineering of virus producer cell lines. The recombinant GPI protein CD59his was purified via fast protein liquid chromatography and associated with concentrated virus stock in a controlled incubation procedure. Reaction mixtures were purified in order to remove nonassociated GPI protein and endogenous protein. Analysis of samples by immunoblotting revealed that CD59his was only detectable in the presence of viral particles. From this, we conclude that CD59his could be stably associated with retroviral particles. In addition, we demonstrated by flow cytometry that virus particles remain infectious after these procedures. As well as suggesting a novel possibility for interaction between enveloped virus and host, we believe that the stable association of recombinant GPI proteins to retroviral particles can be developed into an important tool for both research and clinical applications, especially in the fields of gene therapy and vaccine development.
Metzner
Christoph
C
Institute of Virology, University of Veterinary Medicine, Veterinärplatz 1, A-1210 Vienna, Austria. christoph.metzner@vu-wien.ac.at
Mostegl
Meike M
MM
Günzburg
Walter H
WH
Salmons
Brian
B
Dangerfield
John A
JA
D000818:Animals; D001483:Base Sequence; D018957:CD59 Antigens / Q000235:genetics / Q000378:metabolism*; D002460:Cell Line; D017931:DNA Primers / Q000235:genetics; D017261:Glycosylphosphatidylinositols / Q000378:metabolism*; D006367:HeLa Cells; D006801:Humans; D016086:Lentivirus / Q000378:metabolism; D009052:Leukemia Virus, Murine / Q000378:metabolism; D051379:Mice; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D012190:Retroviridae / Q000378:metabolism*; D014771:Virion / Q000378:metabolism; D014773:Virology / Q000379:methods
D018957:CD59 Antigens; D017931:DNA Primers; D017261:Glycosylphosphatidylinositols; D011994:Recombinant Proteins; C051590:CD59 protein, human