Novel interferon-beta-induced gene expression in peripheral blood cells.
Type I IFNs are used for treating viral, neoplastic, and inflammatory disorders. The protein products encoded by IFN-stimulated genes (ISGs) likely mediate clinical effects of IFN in patients. Macroarray assays, used for studying ISG induction in IFN-treated patients, comprise genes identified predominantly through analysis of long-term cell lines. To discover genes induced selectively by IFN-beta in PBMC, we exposed whole blood to physiological concentrations of IFN-beta. PBMC were prepared, and RNA was extracted, reverse-transcribed, and hybridized to cDNA microarrays, and microarray analysis identified 39 ISGs and 20 IFN-repressed genes (IRGs). Thirty-three ISGs were known previously, and six ISGs were novel. New ISGs included GTP cyclohydrolase 1; hypothetical protein LOC129607; hypothetical protein FLJ38348; leucine aminopeptidase 3; squalene epoxidase; and GTP-binding protein overexpressed in skeletal muscle. Twenty IRGs included IL-1beta and CXCL8, which had been identified earlier. CXCL1 was a novel IRG identified in the current study. PCR analysis demonstrated the regulation of six novel ISGs and CXCL1 as an IRG in PBMC and astrocytoma cells. Results were validated using RNA obtained ex vivo from blood of patients after injection with IFN-beta. Identification of new ISGs and IRGs in primary PBMC will enhance macroarray assays for monitoring IFN responsiveness.
Rani
M R Sandhya
MR
Neuroinflammation Research Center, Department of Neurosciences, Lerner Research Institute, NC30, Mellen Center for MS Treatment and Research, Cleveland Clinic, Cleveland, OH 44195, USA.
Shrock
Jennifer
J
Appachi
Swathi
S
Rudick
Richard A
RA
Williams
Bryan R G
BR
Ransohoff
Richard M
RM
D000970:Antineoplastic Agents / Q000494:pharmacology*; D001254:Astrocytoma / Q000378:metabolism / Q000473:pathology; D015415:Biomarkers / Q000378:metabolism*; D001773:Blood Cells / Q000187:drug effects / Q000378:metabolism*; D020869:Gene Expression Profiling; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects*; D006801:Humans; D016899:Interferon-beta / Q000494:pharmacology*; D009103:Multiple Sclerosis / Q000097:blood* / Q000235:genetics / Q000473:pathology; D020411:Oligonucleotide Array Sequence Analysis; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction
D000970:Antineoplastic Agents; D015415:Biomarkers; D012333:RNA, Messenger; D016899:Interferon-beta