title: Lenalidomide overcomes suppression of human natural killer cell anti-tumor functions by neuroblastoma microenvironment-associated IL-6 and TGFβ1.
abstract: BACKGROUND
  Treatment for children with high-risk neuroblastoma with anti-disialoganglioside mAb ch14.18, IL-2, and GM-CSF plus 13-cis-retinoic acid after myeloablative chemotherapy improves survival, but 40 % of patients still relapse during or after this therapy. The microenvironment of high-risk neuroblastoma tumors includes macrophages, IL-6, and TGFβ1. We hypothesized that this microenvironment suppresses anti-tumor functions of natural killer (NK) cells and that lenalidomide, an immune-modulating drug, could overcome suppression.
  
  
  METHODS
  Purified NK cells were cultured with IL-2, neuroblastoma/monocyte-conditioned culture medium (CM), IL-6, TGFβ1, and lenalidomide in various combinations and then characterized using cytotoxicity (direct and antibody-dependent cell-mediated cytotoxicity), cytokine, flow cytometry, and Western blotting assays. Anti-tumor activity of NK cells with lenalidomide, ch14.18, or both was evaluated with a xenograft model of neuroblastoma.
  
  
  RESULTS
  CM from neuroblastoma/monocyte co-cultures contains IL-6 and TGFβ1 that suppress IL-2 activation of NK cell cytotoxicity and IFNγ secretion. IL-6 and TGFβ1 activate the STAT3 and SMAD2/3 pathways in NK cells and suppress IL-2 induction of cytotoxicity, granzymes A and B release, perforin expression, and IFNγ secretion. Lenalidomide blocks IL-6 and TGFβ1 activation of these signaling pathways and inhibits their suppression of NK cells. Neuroblastoma cells in NOD/SCID mice exhibit activated STAT3 and SMAD2/3 pathways. Their growth is most effectively inhibited by co-injected peripheral blood mononuclear cells (PBMC) containing NK cells when mice are treated with both ch14.18 and lenalidomide.
  
  
  CONCLUSIONS
  Immunotherapy with anti-tumor cell antibodies may be improved by lenalidomide, which enhances activation of NK cells and inhibits their suppression by IL-6 and TGFβ1.
authors: Division of Hematology/Oncology, Children's Hospital Los Angeles, 4650 Sunset Boulevard, Mailstop #57, Los Angeles, CA, 90027, USA.
authors: Yibing
authors: Y
authors: Xu
authors: Jianping
authors: J
authors: Sun
authors: Michael A
authors: MA
authors: Sheard
authors: Hung C
authors: HC
authors: Tran
authors: Zesheng
authors: Z
authors: Wan
authors: Wei Yao
authors: WY
authors: Liu
authors: Shahab
authors: S
authors: Asgharzadeh
authors: Richard
authors: R
authors: Sposto
authors: Hong Wei
authors: HW
authors: Wu
authors: Robert C
authors: RC
authors: Seeger
mesh_terms: D000818:Animals; D000920:Antibody-Dependent Cell Cytotoxicity; D017077:Culture Media, Conditioned; D005260:Female; D006801:Humans; D015850:Interleukin-6 / Q000276:immunology / Q000494:pharmacology*; D007694:Killer Cells, Natural / Q000187:drug effects* / Q000276:immunology; D000077269:Lenalidomide; D051379:Mice; D016688:Mice, Inbred NOD; D016513:Mice, SCID; D009447:Neuroblastoma / Q000188:drug therapy* / Q000276:immunology* / Q000378:metabolism; D013792:Thalidomide / Q000031:analogs & derivatives* / Q000494:pharmacology; D053773:Transforming Growth Factor beta1 / Q000276:immunology / Q000494:pharmacology*; D059016:Tumor Microenvironment / Q000187:drug effects / Q000276:immunology
chemical_list: D017077:Culture Media, Conditioned; D015850:Interleukin-6; D053773:Transforming Growth Factor beta1; D013792:Thalidomide; D000077269:Lenalidomide
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