Genome-wide analysis displays marked induction of EBI3/IL-27B in IL-18-activated AML-derived KG1 cells: critical role of two kappaB binding sites in the human EBI3 promotor.
The cell line KG1 is derived from a patient with acute myeloid leukemia. Activation of KG1 cells by interleukin (IL)-18 is associated with induction of key Th1 signature parameters such as T-bet and interferon-gamma. Here we set out to characterize the genome-wide mRNA expression profile under the condition of short-term stimulation (4h) with IL-18 using the Affymetrix GeneChip((R)) Array System. Besides the chemokines CXCL10, CXCL11, and CCL1 we identified Epstein-Barr virus induced gene-3 (EBI3)/IL-27B as being among those genes that are profoundly upregulated by IL-18 in KG1 cells. Thorough investigation revealed that IL-18-induced EBI3 mRNA efficiently translates into protein. Electromobility shift analysis and mutational analysis of the human EBI3 promoter identified two nuclear factor-kappaB binding sites as being crucial for induction by pro-inflammatory cytokines like IL-18. In addition, we demonstrate that KG1 cells express the Type A IL-27 receptor chain (WSX-1) and display STAT-1, -3 activation as well as induction of SOCS-3 under the influence of IL-27. IL-18 shows therapeutic potential in murine leukemia and is currently being evaluated in phase II clinical trials for the treatment of immunologically sensitive cancers. Since IL-27 mediates anti-cancer bioactivity in animal models, data presented herein may add a novel facet to tumorsuppressive characteristics of IL-18.
Poleganov
Marco A
MA
Pharmazentrum Frankfurt/ZAFES, Klinikum der Johann Wolfgang Goethe-Universität Frankfurt am Main, 60590 Frankfurt am Main, Germany.
Bachmann
Malte
M
Pfeilschifter
Josef
J
Mühl
Heiko
H
D001665:Binding Sites; D045744:Cell Line, Tumor; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects; D015894:Genome, Human / Q000235:genetics*; D006801:Humans; D020381:Interleukin-17 / Q000494:pharmacology; D020382:Interleukin-18 / Q000494:pharmacology*; D007378:Interleukins / Q000235:genetics*; D015470:Leukemia, Myeloid, Acute / Q000235:genetics*; D015778:Minor Histocompatibility Antigens; D016328:NF-kappa B / Q000276:immunology*; D020411:Oligonucleotide Array Sequence Analysis; D011401:Promoter Regions, Genetic / Q000235:genetics*
C097732:EBI3 protein, human; D020381:Interleukin-17; D020382:Interleukin-18; D007378:Interleukins; D015778:Minor Histocompatibility Antigens; D016328:NF-kappa B