Characterization of human non-small cell lung cancer (NSCLC) cell lines for expression of MHC, co-stimulatory molecules and tumor-associated antigens

Lung Cancer. Aug-Sep 2001;33(2-3):181-94. doi: 10.1016/s0169-5002(01)00210-0.

Abstract

A panel of 31 long-term non-small cell lung cancer (NSCLC) cell lines was examined for the expression of protein and/or mRNA transcripts for 11 distinct immune response related molecules or tumor associated antigens (TAA). To assess whether cytokine stimulation might up-regulate expression of the genes of interest, cells were cultured in 500 U/ml of gamma-interferon (gamma-IFN) for 48-72 h prior to analysis. Major histocompatibility complex (MHC) Class I antigens were detected by indirect immunofluorescence and were constitutively expressed on all of the cell lines. The average of the mean fluorescence intensity (MFI) measured 222+/-22. gamma-IFN stimulation produced a significant increase to 482+/-36. For MHC Class II only 7/31 cell lines (23%) exhibited constitutive expression, while gamma-IFN treatment had a dramatic effect and yielded 18/31 (58%) positive cell lines. The co-stimulatory molecules CD80 and CD86 were examined by direct immunofluorescence for cell surface expression and RT-PCR amplification for mRNA. CD80 protein was not detected at all, while an insignificant percentage of cells were positive (mean 2%) for CD86 in all cell lines tested. gamma-IFN had no apparent effect on CD80 or CD86 protein expression. Constitutive CD80 or CD86 mRNA levels were observed in 45 and 61% of the NSCLC lines, respectively. These percentages increased to 77% and 90% with gamma-IFN. Cell surface phenotypic analysis for TAA revealed positive populations in 28/31 cell lines (90%) for Her-2/neu, 18/31 (58%) for CEA and 8/31 (26%) for GD-2, with gamma-IFN having no effect. After gamma-IFN stimulation, RT-PCR amplification for Mage-1, -2, -3 and WT-1 detected mRNA in 33%, 33%, 44% and 70% of the cell lines, respectively. Overall, gamma-IFN stimulation led to the up-regulation of MHC Class I molecules and class II molecules as well as CD80 and CD86 mRNA transcripts. This survey represents the first comprehensive analysis of NSCLC cell lines for a variety of molecules that could play an important role in the generation of an NSCLC anti-tumor CD8+ cytotoxic T lymphocyte (CTL) response.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Antigens, CD / genetics*
  • Antigens, CD / metabolism
  • Antigens, Neoplasm / genetics*
  • Antigens, Neoplasm / metabolism
  • B7-1 Antigen / genetics*
  • B7-1 Antigen / metabolism
  • B7-2 Antigen
  • Carcinoma, Non-Small-Cell Lung / metabolism*
  • DNA Primers / chemistry
  • Flow Cytometry
  • Histocompatibility Antigens / genetics*
  • Histocompatibility Antigens / metabolism
  • Humans
  • Interferon-gamma / pharmacology*
  • Lung Neoplasms / metabolism*
  • Membrane Glycoproteins / genetics*
  • Membrane Glycoproteins / metabolism
  • RNA, Messenger / metabolism
  • Reverse Transcriptase Polymerase Chain Reaction
  • Tumor Cells, Cultured / drug effects*
  • Tumor Cells, Cultured / metabolism
  • Up-Regulation

Substances

  • Antigens, CD
  • Antigens, Neoplasm
  • B7-1 Antigen
  • B7-2 Antigen
  • CD86 protein, human
  • DNA Primers
  • Histocompatibility Antigens
  • Membrane Glycoproteins
  • RNA, Messenger
  • Interferon-gamma