Discordant production of released exogenous protein and infectious virions from retrovirus-packaging cells used for gene transduction.
We examined the correlation between the amount of exogenous protein secreted and that of its viral transcript from PA317 packaging cells transduced with a cytokine gene. Amphotropic packaging cells for the retrovirus expressing murine interleukin-15 (IL-15) gene were cloned, and we examined 22 clones for both the amount of IL-15 secreted with enzyme-linked immunosorbent assay and the IL-15 transcript released as infectious virions by slot blot-hybridization analysis. The study revealed that there was no statistical correlation between them, and suggests that the integrated transcript is used discordantly. Thus, the examination of secreted protein from packaging cells cannot be substituted for laborious assays to determine the amount of virion transcripts when isolating high titer packaging cells.
Tasaki
K
K
Department of Surgery (II), School of Medicine, Chiba University, Japan.
Yoshida
Y
Y
Tagawa
M
M
Takenaga
K
K
Asano
T
T
Ochiai
T
T
Isono
K
K
Kouzu
T
T
Saisho
H
H
Sakiyama
S
S
D000818:Animals; D002999:Clone Cells; D005822:Genetic Vectors; D019409:Interleukin-15 / Q000096:biosynthesis*; D008070:Lipopolysaccharides / Q000494:pharmacology; D008214:Lymphocytes / Q000187:drug effects / Q000276:immunology*; D051379:Mice; D008807:Mice, Inbred BALB C; D016133:Polymerase Chain Reaction; D011994:Recombinant Proteins / Q000096:biosynthesis; D012190:Retroviridae / Q000502:physiology*; D014158:Transcription, Genetic; D014162:Transfection / Q000379:methods*; D014771:Virion / Q000502:physiology*
D019409:Interleukin-15; D008070:Lipopolysaccharides; D011994:Recombinant Proteins