Association of IFN-gamma signal transduction defects with impaired HLA class I antigen processing in melanoma cell lines.
OBJECTIVE
Abnormalities in the constitutive and IFN-γ-inducible HLA class I surface antigen expression of tumor cells is often associated with an impaired expression of components of the antigen processing machinery (APM). Hence, we analyzed whether there exists a link between the IFN-γ signaling pathway, constitutive HLA class I APM component expression, and IFN-γ resistance.


METHODS
The basal and IFN-γ-inducible expression profiles of HLA class I APM and IFN-γ signal transduction cascade components were assessed in melanoma cells by real-time PCR (RT-PCR), Western blot analysis and/or flow cytometry, the integrity of the Janus activated kinase (JAK) 2 locus by comparative genomic hybridization. JAK2 was transiently overexpressed in JAK2(-) cells. The effect of IFN-γ on the cell growth was assessed by XTT [2,3-bis(2-methoxy-4-nitro-S-sulfophenynl)-H-tetrazolium-5-carboxanilide inner salt] assay.


RESULTS
The analysis of 8 melanoma cell lines linked the IFN-γ unresponsiveness of Colo 857 cells determined by lack of inducibility of HLA class I surface expression on IFN-γ treatment to a deletion of JAK2 on chromosome 9, whereas other IFN-γ signaling pathway components were not affected. In addition, the constitutive HLA class I APM component expression levels were significantly reduced in JAK2(-) cells. Furthermore, JAK2-deficient cells were also resistant to the antiproliferative effect of IFN-γ. Transfection of wild-type JAK2 into JAK2(-) Colo 857 not only increased the basal APM expression but also restored their IFN-γ sensitivity.


CONCLUSIONS
Impaired JAK2 expression in melanoma cells leads to reduced basal expression of MHC class I APM components and impairs their IFN-γ inducibility, suggesting that malfunctional IFN-γ signaling might cause HLA class I abnormalities.
Martin-Luther-Universitat Halle-Wittenberg, Institute of Medical Immunology, Halle, Saale, Germany.
Annedore
A
Respa
Juergen
J
Bukur
Soldano
S
Ferrone
Graham
G
Pawelec
Yingdong
Y
Zhao
Ena
E
Wang
Francesco M
FM
Marincola
Barbara
B
Seliger
D017951:Antigen Presentation / Q000235:genetics*; D045744:Cell Line, Tumor; D055028:Comparative Genomic Hybridization; D004351:Drug Resistance / Q000235:genetics; D015395:Histocompatibility Antigens Class I / Q000276:immunology / Q000378:metabolism; D006801:Humans; D007371:Interferon-gamma / Q000378:metabolism* / Q000494:pharmacology; D053614:Janus Kinase 2 / Q000235:genetics / Q000378:metabolism / Q000502:physiology; D008545:Melanoma / Q000235:genetics* / Q000276:immunology* / Q000378:metabolism; D010766:Phosphorylation; D050794:STAT1 Transcription Factor / Q000378:metabolism; D015398:Signal Transduction / Q000235:genetics; D012878:Skin Neoplasms / Q000235:genetics* / Q000276:immunology* / Q000378:metabolism; D014162:Transfection
D015395:Histocompatibility Antigens Class I; D050794:STAT1 Transcription Factor; C494079:STAT1 protein, human; D007371:Interferon-gamma; D053614:Janus Kinase 2
Nat Rev Immunol. 2008 Jul;8(7):559-68
18575461
J Immunol. 2005 Apr 1;174(7):3948-58
15778351
Mol Cancer Res. 2007 Dec;5(12):1331-41
18171991
Nat Rev Immunol. 2006 Nov;6(11):836-48
17063185
Scand J Immunol. 1999 Sep;50(3):242-9
10447932
Cancer Res. 2000 Oct 15;60(20):5789-96
11059775
J Interferon Cytokine Res. 2007 Mar;27(3):181-9
17348816
Curr Med Chem. 2007;14(6):667-79
17346154
Nat Rev Cancer. 2004 Feb;4(2):97-105
14964307
Int J Cancer. 2000 Mar 1;85(5):720-5
10699955
Oncogene. 2002 Apr 11;21(16):2504-12
11971185
Gut. 2010 Jul;59(7):926-33
20581241
Melanoma Res. 2003 Jun;13(3):219-29
12777975
Proc Natl Acad Sci U S A. 1998 Jun 23;95(13):7556-61
9636188
BMC Cancer. 2007 Feb 23;7:34
17319941
J Immunol Methods. 2005 Apr;299(1-2):139-51
15896802
Adv Cancer Res. 2008;101:249-76
19055946
Tissue Antigens. 2005 Sep;66(3):185-94
16101829
Curr Opin Biotechnol. 2008 Feb;19(1):26-9
18053704
Br J Haematol. 1994 Dec;88(4):794-802
7819104
Blood. 2008 May 1;111(9):4809-12
18160671
Cancer Res. 2008 Jan 1;68(1):122-31
18172304
Immunogenetics. 2007 Feb;59(2):123-33
17180681
Oncogene. 2002 Feb 14;21(8):1251-62
11850845
Oncogene. 2000 May 15;19(21):2505-10
10851049
Tissue Antigens. 2003 Nov;62(5):385-93
14617045
Curr Med Chem. 2007;14(12):1279-89
17504213
Clin Cancer Res. 2007 Sep 1;13(17):5010-9
17785551
J Immunol. 1986 Oct 1;137(7):2299-306
3760563
J Immunol. 2004 Jul 15;173(2):1103-10
15240699
J Biol Chem. 1997 Nov 7;272(45):28779-85
9353349
Oncogene. 2000 Nov 20;19(49):5662-79
11114747