Phenotypic and functional characteristics of blood natural killer cells from melanoma patients at different clinical stages

PLoS One. 2013 Oct 18;8(10):e76928. doi: 10.1371/journal.pone.0076928. eCollection 2013.

Abstract

Melanomas are aggressive skin tumors characterized by high metastatic potential. Immunotherapy is a valuable alternative for metastatic melanoma patients resistant to chemotherapy. Natural Killer (NK) cells are efficient anti-tumor cytotoxic effectors. We previously showed that blood NK cells from stage IV metastatic melanoma patients display decreased NK receptors and that chemotherapy modifies the functional status of blood NK cells. To investigate the role of NK cells along melanoma progression, we have here studied NK cells from patients at different stages of the disease. First, we showed that ex vivo NK cells from certain stage III-IV patients displayed low degranulation potential. Using a dynamic label-free assay, we found that immunoselected IL-2 activated blood NK cells from patients efficiently lysed melanoma cells through NKp46 and NKG2D receptors, independently to the clinical stage. Moreover, the ex vivo phenotype of circulating NK cells from 33 patients (stage I to IV) was extensively analyzed. NK cells from patients displayed higher variability in the percentages of Natural Cytotoxicity Receptors (NCR) and Natural Killer Group 2D (NKG2D) receptor expression compared to donor NK cells. The main defect was the decreased expression of NCR1 (NKp46) by NK cells from metastatic patients. Interestingly, we found a positive correlation between the NK cell percentages of NKp46 and the duration of stage IV in melanoma patients. Finally, we showed that NK cells infiltrated primary melanomas and displayed a predominant peritumoral distribution. These results are new arguments for the development of NK-based therapies in melanoma patients.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adult
  • Aged
  • Aged, 80 and over
  • Cell Degranulation / immunology
  • Cell Line, Tumor
  • Cells, Cultured
  • Coculture Techniques
  • Cytotoxicity, Immunologic / immunology
  • Female
  • Flow Cytometry
  • Humans
  • Immunohistochemistry
  • Immunophenotyping
  • Interleukin-2 / immunology
  • Interleukin-2 / pharmacology
  • K562 Cells
  • Killer Cells, Natural / immunology*
  • Killer Cells, Natural / metabolism
  • Lymphocyte Activation / drug effects
  • Lymphocyte Activation / immunology
  • Male
  • Melanoma / blood
  • Melanoma / immunology*
  • Melanoma / pathology
  • Middle Aged
  • NK Cell Lectin-Like Receptor Subfamily K / immunology
  • NK Cell Lectin-Like Receptor Subfamily K / metabolism
  • Natural Cytotoxicity Triggering Receptor 1 / immunology
  • Natural Cytotoxicity Triggering Receptor 1 / metabolism
  • Neoplasm Staging
  • Skin Neoplasms / blood
  • Skin Neoplasms / immunology*
  • Skin Neoplasms / pathology
  • Young Adult

Substances

  • Interleukin-2
  • KLRK1 protein, human
  • NCR1 protein, human
  • NK Cell Lectin-Like Receptor Subfamily K
  • Natural Cytotoxicity Triggering Receptor 1

Supplementary concepts

  • Melanoma, Cutaneous Malignant

Grant support

Grant Support: This work was supported by grants from INSERM, l'Association pour la Recherche contre le Cancer (ARC, 3964), la Société Française de Dermatologie (SFD, grant for Dr JC), la Ligue Nationale contre le Cancer (grant to AC, Comité Ile de France), l'Institut National du Cancer (Cancéropole Ile de France), la Fondation de France (Grant 20363 to AC). GF had a grant from Canceropole IdF and Ligue Nationale contre le Cancer. MM is recipient of a PhD grant from Ligue Nationale contre le Cancer. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.