Quantification of HIV-based lentiviral vectors: influence of several cell type parameters on vector infectivity.
Arch Virol
; 157(2): 217-23, 2012 Feb.
Article
en En
| MEDLINE
| ID: mdl-22042211
ABSTRACT
A human immunodeficiency virus type (HIV-1)-based lentiviral vector pseudotyped with the vesicular stomatitis virus envelope glycoprotein and encoding the GFP reporter gene was used to evaluate different methods of lentiviral vector titration. GFP expression, viral DNA quantification and the efficiency of vector DNA integration were assayed after infection of conventional HIV-1-permissive cell lines and human primary adult fibroblasts with the vector. We found that vector titers based on GFP expression determined by flow cytometry may vary by more than 50-fold depending on the cell type and the promoter-cell combination used. Interestingly, we observed that the viral integration process in primary HDFa cells was significantly more efficient compared to that in SupT1 or 293T cells. We propose that determination of the amount of integrated viral DNA by quantitative PCR be used in combination with the reporter gene expression assay.
Texto completo:
1
Colección:
01-internacional
Base de datos:
MEDLINE
Asunto principal:
VIH-1
/
Integración Viral
/
Técnicas de Transferencia de Gen
/
Vectores Genéticos
Límite:
Humans
Idioma:
En
Revista:
Arch Virol
Año:
2012
Tipo del documento:
Article
País de afiliación:
Francia