Cell-type and donor-specific transcriptional responses to interferon-alpha. Use of customized gene arrays.
A sensitive, specific, reproducible, robust, and cost-effective customized cDNA array system based on established nylon membrane technology has been developed for convenient multisample expression profiling for several hundred genes of choice. The genes represented are easily adjusted (depending on the availability of corresponding cDNAs) and the method is accordingly readily applicable to a wide variety of systems. Here we have focused on the expression profiles for interferon-alpha2a, the most widely used interferon for the treatment of viral hepatitis and malignancies, in primary cells (peripheral blood mononuclear cells, T cells, and dendritic cells) and cell lines (Kit255, HT1080, HepG2, and HuH7). Of 150 genes studied, only six were consistently induced in all cell types and donors, whereas 74 genes were induced in at least one cell type. IRF-7 was identified as the only gene exclusively induced in the hematopoietic cells. No gene was exclusively induced in the nonhematopoietic cell lines. In T cells 12, and in dendritic cells, 25 genes were induced in all donors whereas 45 and 42 genes, respectively, were induced in at least one donor. The data suggest that signaling through IFN-alpha2 can be substantially modulated to yield significant cell-type and donor-specific qualitative and quantitative differences in gene expression in response to this cytokine under highly standardized conditions.
Schlaak
Joerg F
JF
Cancer Research UK, London Research Institute, 44 Lincoln's Inn Fields, London WC2A 3PX, UK. joerg.schlaak@uni-essen.de
Hilkens
Catharien M U
CM
Costa-Pereira
Ana P
AP
Strobl
Birgit
B
Aberger
Fritz
F
Frischauf
Anna-Maria
AM
Kerr
Ian M
IM
D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000378:metabolism; D019992:Databases as Topic; D003713:Dendritic Cells / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D005821:Genetic Techniques; D006801:Humans; D016898:Interferon-alpha / Q000737:chemistry* / Q000378:metabolism*; D007963:Leukocytes, Mononuclear / Q000378:metabolism; D009693:Nucleic Acid Hybridization; D020411:Oligonucleotide Array Sequence Analysis; D016133:Polymerase Chain Reaction; D011485:Protein Binding; D012313:RNA / Q000378:metabolism; D012260:Ribonucleases / Q000378:metabolism; D012680:Sensitivity and Specificity; D015398:Signal Transduction; D013601:T-Lymphocytes / Q000378:metabolism; D014158:Transcription, Genetic; D015854:Up-Regulation
D018076:DNA, Complementary; D016898:Interferon-alpha; D012313:RNA; D012260:Ribonucleases