Journal of Leukocyte Biology
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Originally published online as doi:10.1189/jlb.0507273 on August 20, 2007

Published online before print August 20, 2007
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(Journal of Leukocyte Biology. 2007;82:1353-1360.)
© 2007 by Society for Leukocyte Biology

Novel interferon-β-induced gene expression in peripheral blood cells

M. R. Sandhya Rani*, Jennifer Shrock*, Swathi Appachi*,1, Richard A. Rudick*,{dagger}, Bryan R. G. Williams{ddagger} and Richard M. Ransohoff*,{dagger},2

* Neuroinflammation Research Center, Department of Neurosciences, Lerner Research Institute, and
{dagger} Mellen Center for MS Treatment and Research, Cleveland Clinic, Cleveland, Ohio, USA; and
{ddagger} Monash Institute of Medical Research, Monash University, Clayton, Victoria, Australia

2 Correspondence: Neuroinflammation Research Center, Department of Neurosciences, Lerner Research Institute, NC30, Mellen Center for MS Treatment and Research, Cleveland Clinic, 9500 Euclid Avenue, Cleveland, OH 44195, USA. E-mail: ransohr{at}ccf.org

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-β in PBMC, we exposed whole blood to physiological concentrations of IFN-β. 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-1β 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-β. Identification of new ISGs and IRGs in primary PBMC will enhance macroarray assays for monitoring IFN responsiveness.

Key Words: microarray • IFN-stimulated genes • IFN-repressed genes • multiple sclerosis






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