IL-12 Directs Further Maturation of Ex Vivo Differentiated NK Cells With Improved Therapeutic Potential

PLoS One. 2014 Jan 31;9(1):e87131. doi: 10.1371/journal.pone.0087131. eCollection 2014.

Abstract

The possibility to modulate ex vivo human NK cell differentiation towards specific phenotypes will contribute to a better understanding of NK cell differentiation and facilitate tailored production of NK cells for immunotherapy. In this study, we show that addition of a specific low dose of IL-12 to an ex vivo NK cell differentiation system from cord blood CD34(+) stem cells will result in significantly increased proportions of cells with expression of CD62L as well as KIRs and CD16 which are preferentially expressed on mature CD56(dim) peripheral blood NK cells. In addition, the cells displayed decreased expression of receptors such as CCR6 and CXCR3, which are typically expressed to a lower extent by CD56(dim) than CD56(bright) peripheral blood NK cells. The increased number of CD62L and KIR positive cells prevailed in a population of CD33(+)NKG2A(+) NK cells, supporting that maturation occurs via this subtype. Among a series of transcription factors tested we found Gata3 and TOX to be significantly downregulated, whereas ID3 was upregulated in the IL-12-modulated ex vivo NK cells, implicating these factors in the observed changes. Importantly, the cells differentiated in the presence of IL-12 showed enhanced cytokine production and cytolytic activity against MHC class I negative and positive targets. Moreover, in line with the enhanced CD16 expression, these cells exhibited improved antibody-dependent cellular cytotoxicity for B-cell leukemia target cells in the presence of the clinically applied antibody rituximab. Altogether, these data provide evidence that IL-12 directs human ex vivo NK cell differentiation towards more mature NK cells with improved properties for potential cancer therapies.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antibodies, Monoclonal, Murine-Derived / immunology
  • Antibodies, Monoclonal, Murine-Derived / pharmacology
  • Antibody-Dependent Cell Cytotoxicity / drug effects
  • Antibody-Dependent Cell Cytotoxicity / immunology*
  • Antigens, CD34 / immunology
  • Antigens, CD34 / metabolism
  • Antineoplastic Agents / immunology
  • Antineoplastic Agents / pharmacology
  • Cell Differentiation / drug effects
  • Cell Differentiation / genetics
  • Cell Differentiation / immunology*
  • Cell Line, Tumor
  • Cells, Cultured
  • Dose-Response Relationship, Drug
  • Fetal Blood / cytology
  • Fetal Blood / immunology
  • Fetal Blood / metabolism
  • Flow Cytometry
  • GATA3 Transcription Factor / genetics
  • GATA3 Transcription Factor / immunology
  • Hematopoietic Stem Cells / drug effects
  • Hematopoietic Stem Cells / immunology
  • Hematopoietic Stem Cells / metabolism
  • High Mobility Group Proteins / genetics
  • High Mobility Group Proteins / immunology
  • Humans
  • Immunotherapy, Adoptive / methods
  • Inhibitor of Differentiation Proteins / genetics
  • Inhibitor of Differentiation Proteins / immunology
  • Interleukin-2 / immunology*
  • Interleukin-2 / pharmacology
  • K562 Cells
  • Killer Cells, Natural / drug effects
  • Killer Cells, Natural / immunology*
  • Killer Cells, Natural / metabolism
  • L-Selectin / immunology
  • L-Selectin / metabolism
  • Neoplasm Proteins / genetics
  • Neoplasm Proteins / immunology
  • Receptors, CCR6 / immunology
  • Receptors, CCR6 / metabolism
  • Receptors, CXCR3 / immunology
  • Receptors, CXCR3 / metabolism
  • Receptors, IgG / immunology
  • Receptors, IgG / metabolism
  • Receptors, KIR / immunology
  • Receptors, KIR / metabolism
  • Reverse Transcriptase Polymerase Chain Reaction
  • Rituximab

Substances

  • Antibodies, Monoclonal, Murine-Derived
  • Antigens, CD34
  • Antineoplastic Agents
  • CCR6 protein, human
  • CXCR3 protein, human
  • GATA3 Transcription Factor
  • Gata3 protein, mouse
  • High Mobility Group Proteins
  • Inhibitor of Differentiation Proteins
  • Interleukin-2
  • Neoplasm Proteins
  • Receptors, CCR6
  • Receptors, CXCR3
  • Receptors, IgG
  • Receptors, KIR
  • TOX protein, human
  • L-Selectin
  • ID3 protein, human
  • Rituximab

Grant support

This work was supported by Marie Curie research training network grants of the EC to EH (MRTN-CT-2005-019248 and FP7-People-2012-ITN-317013). DL was a Marie Curie fellow in the network MRTN-CT-2005-019248. The funder had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.