IFN-gamma impairs release of IL-8 by IL-1beta-stimulated A549 lung carcinoma cells.
BACKGROUND
Production of interferon (IFN)-gamma is key to efficient anti-tumor immunity. The present study was set out to investigate effects of IFNgamma on the release of the potent pro-angiogenic mediator IL-8 by human A549 lung carcinoma cells.


METHODS
A549 cells were cultured and stimulated with interleukin (IL)-1beta alone or in combination with IFNgamma. IL-8 production by these cells was analyzed with enzyme linked immuno sorbent assay (ELISA). mRNA-expression was analyzed by real-time PCR and RNase protection assay (RPA), respectively. Expression of inhibitor-kappa Balpha, cellular IL-8, and cyclooxygenase-2 was analyzed by Western blot analysis.


RESULTS
Here we demonstrate that IFNgamma efficiently reduced IL-8 secretion under the influence of IL-1beta. Surprisingly, real-time PCR analysis and RPA revealed that the inhibitory effect of IFNgamma on IL-8 was not associated with significant changes in mRNA levels. These observations concurred with lack of a modulatory activity of IFNgamma on IL-1beta-induced NF-kappaB activation as assessed by cellular IkappaB levels. Moreover, analysis of intracellular IL-8 suggests that IFNgamma modulated IL-8 secretion by action on the posttranslational level. In contrast to IL-8, IL-1beta-induced cyclooxygenase-2 expression and release of IL-6 were not affected by IFNgamma indicating that modulation of IL-1beta action by this cytokine displays specificity.


CONCLUSIONS
Data presented herein agree with an angiostatic role of IFNgamma as seen in rodent models of solid tumors and suggest that increasing T helper type 1 (Th1)-like functions in lung cancer patients e.g. by local delivery of IFNgamma may mediate therapeutic benefit via mechanisms that potentially include modulation of pro-angiogenic IL-8.
Boost
Kim A
KA
Klinik für Anaesthesiologie, Ludwig-Maximilians-University Munich, Marchioninistr. 15, 81377 Munich, Germany. kim.boost@med.uni-muenchen.de
Sadik
Christian D
CD
Bachmann
Malte
M
Zwissler
Bernhard
B
Pfeilschifter
Josef
J
Mühl
Heiko
H
D045744:Cell Line, Tumor; D051546:Cyclooxygenase 2 / Q000096:biosynthesis; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D020672:I-kappa B Proteins / Q000378:metabolism; D007371:Interferon-gamma / Q000276:immunology*; D053583:Interleukin-1beta / Q000037:antagonists & inhibitors / Q000494:pharmacology; D015850:Interleukin-6 / Q000276:immunology; D016209:Interleukin-8 / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology*; D008175:Lung Neoplasms / Q000098:blood supply / Q000235:genetics / Q000276:immunology*; D009389:Neovascularization, Pathologic / Q000276:immunology; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics
D020672:I-kappa B Proteins; D053583:Interleukin-1beta; D015850:Interleukin-6; D016209:Interleukin-8; D012333:RNA, Messenger; D007371:Interferon-gamma; D051546:Cyclooxygenase 2
J Nucl Med. 1997 Sep;38(9):1475-81
9293812
Clin Cancer Res. 2003 Feb;9(2):729-37
12576442
Clin Cancer Res. 1997 Dec;3(12 Pt 2):2682-6
10068274
Am J Respir Cell Mol Biol. 1999 May;20(5):964-75
10226066
Am J Respir Cell Mol Biol. 2005 Jun;32(6):540-7
15746434
Lung Cancer. 2005 Dec;50(3):309-17
16125276
Eur J Cancer. 2006 Apr;42(6):768-78
16510280
Curr Pharm Des. 2006;12(24):3101-15
16918437
Mutagenesis. 2006 Jul;21(4):225-36
16870698
Cancer Metastasis Rev. 2006 Sep;25(3):307-13
17029030
Lung Cancer. 2007 Apr;56(1):25-33
17175059
BMC Infect Dis. 2008;8:17
18267006
Hum Gene Ther. 2000 Jan 20;11(2):247-61
10680839
Lab Invest. 2000 May;80(5):617-53
10830774
Am J Respir Crit Care Med. 2000 Nov;162(5):1957-63
11069840
Lung Cancer. 2001 Jan;31(1):25-30
11162863
Cytokine Growth Factor Rev. 2002 Apr;13(2):95-109
11900986
J Immunol. 2002 Jul 1;169(1):469-75
12077278
Cancer Immunol Immunother. 2002 Nov;51(10):521-31
12384803
Cancer Sci. 2003 Mar;94(3):244-52
12824917
Int Immunopharmacol. 2003 Sep;3(9):1247-55
12890422
FASEB J. 2004 Jan;18(1):161-3
14597552
Science. 1992 Dec 11;258(5089):1798-801
1281554
J Biol Chem. 1993 Mar 15;268(8):5834-9
7680648
Am J Respir Cell Mol Biol. 1994 Feb;10(2):202-6
8110475
Eur J Cancer. 1994;30A(1):11-5
8142151
J Exp Med. 1994 May 1;179(5):1409-15
7513008
Int J Radiat Oncol Biol Phys. 1995 Jan 1;31(1):93-101
7995773
J Leukoc Biol. 1995 Jun;57(6):929-35
7790776
Blood. 1996 Mar 15;87(6):2095-147
8630372
J Clin Invest. 1996 Jun 15;97(12):2792-802
8675690
Melanoma Res. 1996 Aug;6(4):307-11
8873050
J Exp Med. 1996 Sep 1;184(3):981-92
9064358
Lancet. 1997 May 24;349(9064):1513-5
9167461
Int J Cancer. 1998 Jul 3;77(1):7-12
9639386