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IL-2 and IL-12 alter NK cell responsiveness to IFN-gamma-inducible protein 10 by down-regulating CXCR3 expression
Deborah L Hodge1, William B Schill, Ji Ming Wang
1Laboratories of. Experimental Immunology and Molecular Immunoregulation, Center for Cancer Research, National Cancer Institute, Frederick, MD 21702, USA.
Abstract:
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-gamma-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.
Insights
Interleukin-2 (IL-2) and Interleukin-12 (IL-12) treatment rapidly reduces chemokine receptor CXCR3 expression in natural killer (NK) cells. This down-regulation impairs NK cell migration, highlighting a direct mechanism for cytokine-mediated immune cell movement control.
Area of Science:
- Immunology
- Molecular Biology
- Cell Biology
Background:
- Cytokine stimulation profoundly impacts natural killer (NK) cell functions, including cytotoxicity, proliferation, and migration.
- Understanding the molecular basis of these NK cell responses to cytokines like IL-2, IL-12, and IL-18 is crucial for immune modulation.
Purpose of the Study:
- To investigate the molecular mechanisms underlying cytokine-induced alterations in NK cell responses.
- To identify specific gene expression changes in NK92 cells stimulated with IL-2, IL-12, and IL-18.
- To examine the regulation of chemokine receptor expression and its impact on NK cell chemotaxis.
Main Methods:
- Microarray analysis to compare gene expression profiles in unstimulated and cytokine-stimulated NK92 cells.
- RNase protection assays and RT-PCR to validate microarray findings and assess mRNA expression.
- Analysis of CXCR3 receptor mRNA and surface expression in primary NK cells following IL-2 and IL-12 treatment.
- Investigation of transcriptional and posttranscriptional regulation of CXCR3 mRNA decay.
Main Results:
- Microarray analysis revealed significant changes in genes related to cell cycle, signal transduction, and chemotaxis.
- RNase protection assays confirmed alterations in nine genes and detected changes in CCR2 and CXCR3 mRNA expression.
- IL-2 and IL-12 treatment led to a significant decrease in CXCR3 receptor mRNA and surface expression in primary NK cells.
- Reduced CXCR3 expression correlated with diminished NK cell chemotaxis towards IFN-gamma-inducible protein-10.
- CXCR3 mRNA decline was attributed to both transcriptional and posttranscriptional regulatory mechanisms.
Conclusions:
- IL-2 and IL-12 directly down-regulate CXCR3 chemokine receptor expression in NK cells.
- This rapid downregulation of CXCR3 impacts NK cell migratory capacity.
- Cytokine-mediated regulation of chemokine receptor expression offers a direct mechanism to control NK cell migration.
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