The potential of retroviral vectors to cotransfer human endogenous retroviruses (HERVs) from human packaging cell lines.
Using a versatile and highly sensitive retroviral microarray, we have investigated particle preparations from three different human packaging cell lines harboring retroviral vector systems based on human immunodeficiency virus (HIV) and murine leukemia virus (MLV). 293Rev/Gag/Pol(i) cells inducibly express high titers of HIV-derived particles for packaging of HIV vectors. The Phoenix-GP and the Anjou 65 cell lines constitutively express MLV vector particles. We compared the transcription profiles of human endogenous retroviruses (HERVs) in all cell lines with the HERV sequences present in the particles. In addition, the influence of the transfected vector plasmid on the copackaging of HERVs was investigated. All particle preparations showed a defined pattern of endogenous retroviral sequences that differed from the cellular HERV expression pattern. HERV transcripts were observed in the particle preparations independent of whether a vector construct was coexpressed or not. Furthermore, our results suggest that particle preparations are frequently contaminated by cellular vesicles (exosomes) containing cellular RNAs including HERV transcripts.
Zeilfelder
Udo
U
Medical Clinic III, Faculty of Clinical Medicine Mannheim, University of Heidelberg, 68305 Mannheim, Germany.
Frank
Oliver
O
Sparacio
Sandra
S
Schön
Ulrike
U
Bosch
Valerie
V
Seifarth
Wolfgang
W
Leib-Mösch
Christine
C
D000818:Animals; D002460:Cell Line; D020077:Endogenous Retroviruses / Q000235:genetics* / Q000302:isolation & purification; D020869:Gene Expression Profiling; D015316:Genetic Therapy; D005822:Genetic Vectors; D015497:HIV-1 / Q000235:genetics* / Q000502:physiology; D006801:Humans; D009052:Leukemia Virus, Murine / Q000235:genetics* / Q000502:physiology; D051379:Mice; D020411:Oligonucleotide Array Sequence Analysis; D014162:Transfection; D019065:Virus Assembly