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-Inducible Protein 10 by Down-Regulating CXCR3 Expression1
Laboratories of
* Experimental Immunology and
Molecular Immunoregulation, Center for Cancer Research, National Cancer Institute, Frederick, MD 21702; and
National Fish Health Research Laboratory, U.S. Geological Survey-Leetown Science Center, Kearneysville, WV 25430
Cytokine treatment of NK cells results in alterations in multiple
cellular responses that include cytotoxicity, cytokine production,
proliferation, and chemotaxis. To understand the molecular mechanisms
underlying these responses, microarray analysis was performed and the
resulting gene expression patterns were compared between unstimulated,
IL-2, IL-2 plus IL-12, and IL-2 plus IL-18-stimulated NK92 cells. RNase
protection assays and RT-PCR confirmed microarray predictions
for changes in mRNA expression for nine genes involved in cell cycle
progression, signal transduction, transcriptional activation, and
chemotaxis. Multiprobe RNase protection assay also detected changes in
the expression of CCR2 mRNA, a gene that was not imprinted on
the microarray. We subsequently expanded our search for other chemokine
receptor genes absent from the microarray and found an IL-2- and
IL-12-dependent decrease in CXCR3 receptor mRNA expression in NK92
cells. A detailed analysis of CXCR3 expression in primary NK cells
revealed that an IL-2 and an IL-12 together significantly decreased the
CXCR3 receptor mRNA and receptor surface expression by 6 and 24 h
of treatment, respectively. This decrease in receptor expression was
associated with a significant reduction in chemotaxis in the presence
of IFN-
-inducible protein-10. The decline in CXCR3 mRNA was
due to transcriptional and posttranscriptional mechanisms as the
addition of actinomycin D to IL-2- and IL-12-treated NK92 slightly
altered the half-life of the CXCR3 mRNA. Collectively, these data
suggest that IL-2 and IL-12 directly affect NK cell migratory ability
by rapid and direct down-regulation of chemokine receptor mRNA
expression.
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