In vitro cross-species infections using a caprine arthritis encephalitis lentivirus carrying the GFP marker gene.
A caprine arthritis encephalitis virus (CAEV), carrying the green fluorescent protein (GFP) into the tat region was recently reported [Mselli-Lakhal, L., Guiguen, F., Greenland, T., Mornex, J.F., Chebloune, Y., 2006. Gene transfer system derived from the caprine arthritis-encephalitis lentivirus. J. Virol. Meth. 136, 177-184]. This construct, called pK2EGFPH replicated to titres up to 10(5)IU/ml on infection of caprine cells, and could be concentrated to 10(6)IU/ml by ultracentrifugation. In the present study, the pK2EGFPH construct was characterized better and used in cross-species infection studies. The pK2EGFPH virus could transduce GFP protein expression both to goat synovial membrane cells and to an immortalized goat milk epithelial cell line. The pK2EGFPH infected cells were demonstrated to express both GFP protein and CAEV viral proteins, as demonstrated by radioimmunoprecipitation and multinucleated cell formation. However GFP expression could not be maintained over passages. This vector was used to investigate cross-species infectious potential of CAEV. The bovine cell lines MDBK and GBK were found to be sensitive to infection while the human cell lines Hela, A431 and THP-1 were not. The pK2EGFPH vector should prove useful in studies of CAEV tropism both in vitro and in vivo.
Mselli-Lakhal
Laila
L
UMR 754 INRA/ENVL/UCBL Retrovirus et Pathologie Comparée Université Lyon-1, Bâtiment B, 50, Avenue Tony Garnier, 69366 Lyon Cedex 07, France. lakhal@toulouse.inra.fr
Guiguen
François
F
Greenland
Timothy
T
Mornex
Jean-François
JF
Chebloune
Yahia
Y
D000818:Animals; D016088:Arthritis-Encephalitis Virus, Caprine / Q000235:genetics* / Q000302:isolation & purification / Q000502:physiology*; D002417:Cattle; D002460:Cell Line; D045744:Cell Line, Tumor; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins / Q000096:biosynthesis* / Q000235:genetics; D006801:Humans; D016180:Lentivirus Infections / Q000821:virology; D013045:Species Specificity; D014161:Transduction, Genetic; D014779:Virus Replication
D049452:Green Fluorescent Proteins