Association of IFN-γ signal transduction defects with impaired HLA class I antigen processing in melanoma cell lines

Annedore Respa, Juergen Bukur, Soldano Ferrone, Graham Pawelec, Yingdong Zhao, Ena Wang, Francesco M. Marincola, Barbara Seliger

Research output: Contribution to journalArticle

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Abstract

Purpose: 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. Experimental Design: 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.

Original languageEnglish
Pages (from-to)2668-2678
Number of pages11
JournalClinical Cancer Research
Volume17
Issue number9
DOIs
Publication statusPublished - 1 May 2011
Externally publishedYes

Fingerprint

Histocompatibility Antigens Class I
Antigen Presentation
HLA Antigens
Melanoma
Signal Transduction
Cell Line
Janus Kinase 2
Chromosomes, Human, Pair 9
Comparative Genomic Hybridization
Surface Antigens
Transfection
Real-Time Polymerase Chain Reaction
Flow Cytometry
Research Design
Salts
Western Blotting
Growth

ASJC Scopus subject areas

  • Cancer Research
  • Oncology

Cite this

Association of IFN-γ signal transduction defects with impaired HLA class I antigen processing in melanoma cell lines. / Respa, Annedore; Bukur, Juergen; Ferrone, Soldano; Pawelec, Graham; Zhao, Yingdong; Wang, Ena; Marincola, Francesco M.; Seliger, Barbara.

In: Clinical Cancer Research, Vol. 17, No. 9, 01.05.2011, p. 2668-2678.

Research output: Contribution to journalArticle

Respa, Annedore ; Bukur, Juergen ; Ferrone, Soldano ; Pawelec, Graham ; Zhao, Yingdong ; Wang, Ena ; Marincola, Francesco M. ; Seliger, Barbara. / Association of IFN-γ signal transduction defects with impaired HLA class I antigen processing in melanoma cell lines. In: Clinical Cancer Research. 2011 ; Vol. 17, No. 9. pp. 2668-2678.
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T1 - Association of IFN-γ signal transduction defects with impaired HLA class I antigen processing in melanoma cell lines

AU - Respa, Annedore

AU - Bukur, Juergen

AU - Ferrone, Soldano

AU - Pawelec, Graham

AU - Zhao, Yingdong

AU - Wang, Ena

AU - Marincola, Francesco M.

AU - Seliger, Barbara

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N2 - Purpose: 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. Experimental Design: 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.

AB - Purpose: 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. Experimental Design: 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.

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