IRF1 and NF-kB restore MHC class I-restricted tumor antigen processing and presentation to cytotoxic T cells in aggressive neuroblastoma

PLoS One. 2012;7(10):e46928. doi: 10.1371/journal.pone.0046928. Epub 2012 Oct 5.

Abstract

Neuroblastoma (NB), the most common solid extracranial cancer of childhood, displays a remarkable low expression of Major Histocompatibility Complex class I (MHC-I) and Antigen Processing Machinery (APM) molecules, including Endoplasmic Reticulum (ER) Aminopeptidases, and poorly presents tumor antigens to Cytotoxic T Lymphocytes (CTL). We have previously shown that this is due to low expression of the transcription factor NF-kB p65. Herein, we show that not only NF-kB p65, but also the Interferon Regulatory Factor 1 (IRF1) and certain APM components are low in a subset of NB cell lines with aggressive features. Whereas single transfection with either IRF1, or NF-kB p65 is ineffective, co-transfection results in strong synergy and substantial reversion of the MHC-I/APM-low phenotype in all NB cell lines tested. Accordingly, linked immunohistochemistry expression patterns between nuclear IRF1 and p65 on the one hand, and MHC-I on the other hand, were observed in vivo. Absence and presence of the three molecules neatly segregated between high-grade and low-grade NB, respectively. Finally, APM reconstitution by double IRF1/p65 transfection rendered a NB cell line susceptible to killing by anti MAGE-A3 CTLs, lytic efficiency comparable to those seen upon IFN-γ treatment. This is the first demonstration that a complex immune escape phenotype can be rescued by reconstitution of a limited number of master regulatory genes. These findings provide molecular insight into defective MHC-I expression in NB cells and provide the rational for T cell-based immunotherapy in NB variants refractory to conventional therapy.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antigen Presentation / drug effects
  • Antigen Presentation / immunology*
  • Antigens / genetics
  • Antigens / immunology
  • Antigens / metabolism
  • Antigens, Neoplasm / genetics
  • Antigens, Neoplasm / immunology
  • Antigens, Neoplasm / metabolism
  • Blotting, Western
  • Cell Line, Tumor
  • Histocompatibility Antigens Class I / immunology*
  • Histocompatibility Antigens Class I / metabolism
  • Humans
  • Immunohistochemistry
  • Interferon Regulatory Factor-1 / genetics
  • Interferon Regulatory Factor-1 / immunology*
  • Interferon Regulatory Factor-1 / metabolism
  • Interferon-gamma / immunology
  • Interferon-gamma / pharmacology
  • Jurkat Cells
  • Neoplasm Proteins / genetics
  • Neoplasm Proteins / immunology
  • Neoplasm Proteins / metabolism
  • Neuroblastoma / immunology
  • Neuroblastoma / metabolism
  • Neuroblastoma / pathology
  • RNA Interference
  • Reverse Transcriptase Polymerase Chain Reaction
  • T-Lymphocytes, Cytotoxic / immunology*
  • T-Lymphocytes, Cytotoxic / metabolism
  • Transcription Factor RelA / genetics
  • Transcription Factor RelA / immunology*
  • Transcription Factor RelA / metabolism
  • Transfection
  • Up-Regulation / immunology

Substances

  • Antigens
  • Antigens, Neoplasm
  • Histocompatibility Antigens Class I
  • IRF1 protein, human
  • Interferon Regulatory Factor-1
  • MAGEA3 protein, human
  • Neoplasm Proteins
  • Transcription Factor RelA
  • Interferon-gamma

Grants and funding

Associazione Italiana per la Ricerca sul Cancro (AIRC, Milan, Italy) to D.F. and P.G. and the special grant 5 x 1000 AIRC to F.L. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.