IL-12 directs further maturation of ex vivo differentiated NK cells with improved therapeutic potential

• Published in: PloS one Vol. 9; no. 1; p. e87131
• Main Authors: Lehmann, Dorit, Spanholtz, Jan, Sturtzel, Caterina, Tordoir, Marleen, Schlechta, Bernhard, Groenewegen, Dirk, Hofer, Erhard
• Format: Journal Article
• Language: English
• Published: United States Public Library of Science 31.01.2014; Public Library of Science (PLoS)
• Subjects: 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; Biology; Blood; Cancer; CCR6 protein; CD16 antigen; CD34 antigen; Cell differentiation; Cell Differentiation - drug effects; Cell Differentiation - genetics; Cell Differentiation - immunology; Cell Line, Tumor; Cells, Cultured; Chemokines; Cord blood; CXCR3 protein; Cytokines; Cytolytic activity; Cytotoxicity; Differentiation (biology); Dose-Response Relationship, Drug; Fc receptors; Fetal Blood - cytology; Fetal Blood - immunology; Fetal Blood - metabolism; Flow Cytometry; GATA-3 protein; GATA3 Transcription Factor - genetics; GATA3 Transcription Factor - immunology; Gene expression; Gynecology; Health aspects; 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; Immunotherapy, Adoptive - methods; Inhibitor of Differentiation Proteins - genetics; Inhibitor of Differentiation Proteins - immunology; Interleukin 12; Interleukin-2 - immunology; Interleukin-2 - pharmacology; K562 Cells; Killer cell immunoglobulin-like receptors; Killer cells; Killer Cells, Natural - drug effects; Killer Cells, Natural - immunology; Killer Cells, Natural - metabolism; L-selectin; L-Selectin - immunology; L-Selectin - metabolism; Leukemia; Ligands; Lymphocyte receptors; Lymphocytes B; Major histocompatibility complex; Maturation; Medicine; Monoclonal antibodies; Natural killer cells; Neoplasm Proteins - genetics; Neoplasm Proteins - immunology; NKG2 antigen; Obstetrics; Peripheral blood; Pharmacology; Physiology; Receptors; 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; Stem cells; T cell receptors; Targeted cancer therapy; Thrombosis; Toxicity; Transcription factors; Umbilical cord; Austria
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0087131

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.