Development of a lentiviral vector system to study the role of the Andes virus glycoproteins.
To infect target cells, enveloped viruses use their virion surface proteins to direct cell attachment and subsequent entry via virus-cell membrane fusion. How hantaviruses enter cells has been largely unexplored. To study early steps of Andes virus (ANDV) cell infection, a lentiviral vector system was developed based on a Simian immunodeficiency virus (SIV) vector pseudotyped with the ANDV-Gn/Gc envelope glycoproteins. The incorporation of Gn and Gc onto SIV-derived vector particles was assessed using newly generated monoclonal antibodies against ANDV glycoproteins. In addition, sera of ANDV infected humans were able to block cell entry of the SIV vector pseudotyped with ANDV glycoproteins, suggesting that their antigenic conformation is similar to that in the native virus. The use of such SIV vector pseudotyped with ANDV-Gn/Gc glycoproteins should facilitate studies on ANDV cell entry. Along this line, it was found that depletion of cholesterol from target cells strongly diminished cell infection, indicating a possible role of lipid rafts in ANDV cell entry. The Gn/Gc pseudotyped SIV vector has several advantages, notably high titer vector production and easy quantification of cell infection by monitoring GFP reporter gene expression by flow cytometry. Such pseudotyped SIV vectors can be used to identify functional domains in the Gn/Gc glycoproteins and to screen for potential hantavirus cell entry inhibitors.
Cifuentes-Muñoz
Nicolás
N
Fundación Ciencia para la Vida and Instituto Milenio MIFAB, Av. Zañartu 1482, Santiago, Chile; Universidad Andrés Bello, Santiago, Chile.
Darlix
Jean-Luc
JL
Tischler
Nicole D
ND
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D000914:Antibodies, Viral / Q000276:immunology; D002460:Cell Line; D002522:Chlorocebus aethiops; D005822:Genetic Vectors; D006023:Glycoproteins / Q000235:genetics / Q000502:physiology*; D006476:Orthohantavirus / Q000235:genetics / Q000502:physiology*; D006801:Humans; D021962:Membrane Microdomains / Q000502:physiology; D051379:Mice; D011991:Receptors, Virus / Q000502:physiology; D015302:Simian Immunodeficiency Virus / Q000235:genetics*; D014759:Viral Envelope Proteins / Q000235:genetics / Q000502:physiology*; D053586:Virus Internalization
D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D006023:Glycoproteins; D011991:Receptors, Virus; D014759:Viral Envelope Proteins