Ex vivo activation of CD56(+) immune cells that eradicate neuroblastoma

Cancer Res. 2013 Apr 15;73(8):2608-18. doi: 10.1158/0008-5472.CAN-12-3322. Epub 2013 Feb 25.

Abstract

Despite the use of intensive contemporary multimodal therapy, the overall survival of patients with high-risk neuroblastoma is still less than 50%. Therefore, immunotherapy without cross-resistance and overlapping toxicity has been proposed. In this study, we report the development of a novel strategy to specifically activate and expand human CD56(+) (NCAM1) natural killer (NK) immune cells from normal donors and patients with neuroblastoma. Enriched CD56(+) cells from peripheral blood were mixed with CD56(-) fraction at 1:1 ratio and cultured in the presence of OKT3, interleukin (IL)-2, and -15 for five days and then without OKT3 for 16 more days. The final products contained more than 90% CD56(+) cells and could kill neuroblastoma cells effectively that were originally highly resistant to nonprocessed NK cells. Mechanistically, cytolysis of neuroblastoma was mediated through natural cytotoxicity receptor (NCR), DNAX accessory molecule-1 (DNAM-1; CD226), perforin, and granzyme B. Successful clinical scale-up in a good manufacturing practices (GMP)-compliant bioreactor yielded effector cells that in a neuroblastoma xenograft model slowed tumor growth and extended survival without GVHD. Investigation of CD56(+) cells from patients with neuroblastoma revealed a similar postactivation phenotype and lytic activity. Our findings establish a novel and clinically expedient strategy to generate allogeneic or autologous CD56(+) cells that are highly cytotoxic against neuroblastoma with minimal risk of GVHD.

MeSH terms

  • Animals
  • Antigens, Differentiation, T-Lymphocyte / immunology
  • Antigens, Differentiation, T-Lymphocyte / metabolism
  • CD56 Antigen / metabolism*
  • Cell Culture Techniques / standards
  • Cell Degranulation / immunology
  • Cell Line, Tumor
  • Coculture Techniques
  • Cytotoxicity, Immunologic
  • Disease Models, Animal
  • Graft vs Host Reaction / immunology
  • Humans
  • Killer Cells, Natural / cytology
  • Killer Cells, Natural / immunology*
  • Killer Cells, Natural / metabolism*
  • Leukocytes, Mononuclear / cytology
  • Leukocytes, Mononuclear / immunology
  • Leukocytes, Mononuclear / metabolism
  • Lymphocyte Activation / immunology*
  • Mice
  • Natural Killer T-Cells / cytology
  • Natural Killer T-Cells / immunology
  • Natural Killer T-Cells / metabolism
  • Neuroblastoma / immunology*
  • Neuroblastoma / metabolism*
  • Neuroblastoma / therapy
  • Receptors, Natural Killer Cell / immunology
  • Receptors, Natural Killer Cell / metabolism

Substances

  • Antigens, Differentiation, T-Lymphocyte
  • CD226 antigen
  • CD56 Antigen
  • Receptors, Natural Killer Cell