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Analysis of Human T Cell Activity in an Allogeneic Co-Culture Setting of Pre-Treated Tumor Cells
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Distinct patterns of cytolytic T-cell activation by different tumour cells revealed by Ca2+ signalling and granule

Melissa Frick1, Pierre Mouchacca1, Grégory Verdeil1

  • 1Centre d'Immunologie de Marseille-Luminy, Aix Marseille Université UM2, Inserm, U1104, CNRS UMR7280, Marseille, France.

Immunology
|October 8, 2016
PubMed

Insights

Cytotoxic CD8 T effector cells (CTL) show reduced killing of P1A-expressing melanoma cells compared to mastocytoma cells. This difference is linked to impaired cytotoxic granule release in melanoma, potentially due to lower intercellular adhesion molecule-1 expression.

Area of Science:

  • Immunology
  • Cancer Biology
  • Cellular Immunology

Background:

  • Cancer-germline genes encode antigens targeted by cytotoxic CD8 T effector cells (CTL).
  • Previous studies showed P1A-specific CTL effectively regressed P1A-expressing P815 mastocytoma but were less effective against P1A-expressing melanoma cells.

Purpose of the Study:

  • To analyze in vitro interactions between P1A-specific CTL and mastocytoma or melanoma cells.
  • To investigate the mechanisms underlying differential CTL-mediated tumor cell killing.

Main Methods:

  • In vitro co-culture of P1A-expressing tumor cell lines (mastocytoma and melanoma) with P1A-specific CTL.
  • Video-microscopy to analyze early CTL-target cell interaction events, including cytoplasmic Ca2+ ([Ca2+]i) concentration.
  • Analysis of cytotoxic granule (GZMB-Tom) migration and exocytosis.
  • Assessment of surface molecule expression (intercellular adhesion molecule-1).

Main Results:

  • Mastocytoma cells were more sensitive to CTL-mediated lysis than melanoma cells.
  • Both tumor types induced similar increases in cytoplasmic Ca2+ concentration in CTL.
  • CTL showed delayed cytotoxic granule migration and partially deficient exocytosis towards melanoma cells.
  • Mastocytoma cells expressed intercellular adhesion molecule-1, which was absent on melanoma cells.

Conclusions:

  • Differential sensitivity of tumor cells to CTL killing is not due to initial CTL activation (Ca2+ flux).
  • Impaired cytotoxic granule exocytosis and delayed migration in response to melanoma cells contribute to reduced CTL efficacy.
  • Intercellular adhesion molecule-1 expression on mastocytoma cells may facilitate CTL interaction and enhance tumor cell killing.
Keywords:
cytolytic T lymphocytefluorescent granzyme Bmelanomatumour

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