Retrovirus vector-targeted inducible expression of human beta-interferon gene to B-cells.
We have introduced the human beta-interferon gene with its promoter region into murine B-cell and fibroblast cell lines via a Moloney murine leukemia virus (M-MuLV) vector and have studied the inducible expression of the beta-interferon gene as a function of the various retroviral vector designs. By deleting the enhancer within the 3' viral long terminal repeat (LTR), inserting the human beta-interferon gene, and varying placement of the immunoglobulin heavy chain enhancer, we were able to construct vectors which yielded proviruses with various cell type-specific regulation. One of the vectors (pT154) led to a greater than 21-fold increase in beta-interferon protein synthesis after viral infection in the two B-cell lines analyzed, while no inducibility was seen in the fibroblast cells. The data show that inducible beta-interferon expression within a MuLV vector was highly dependent on the absence of the viral enhancer region in the long terminal repeat and the orientation of the beta-interferon gene within the proviral transcriptional unit; the insertion of the immunoglobulin enhancer elevated both constitutive and (or) inducible expression of beta-interferon in B-cells but inhibited constitutive expression of this gene in fibroblasts.
Engelhardt
J F
JF
Oncology Center, Johns Hopkins University, School of Medicine, Baltimore, Maryland 21205.
Kellum
M J
MJ
Bisat
F
F
Pitha
P M
PM
D000818:Animals; D001402:B-Lymphocytes / Q000187:drug effects / Q000378:metabolism*; D002460:Cell Line; D003001:Cloning, Molecular; D003513:Cycloheximide / Q000494:pharmacology; D004742:Enhancer Elements, Genetic; D005347:Fibroblasts / Q000187:drug effects / Q000378:metabolism; D005822:Genetic Vectors / Q000235:genetics*; D006801:Humans; D007143:Immunoglobulin Heavy Chains / Q000235:genetics*; D007370:Interferon Type I / Q000096:biosynthesis / Q000235:genetics*; D009052:Leukemia Virus, Murine / Q000235:genetics*; D051379:Mice; D008815:Mice, Inbred Strains; D012091:Repetitive Sequences, Nucleic Acid; D014162:Transfection
D007143:Immunoglobulin Heavy Chains; D007370:Interferon Type I; D003513:Cycloheximide