Generation of tumor antigen-specific T cell lines from pediatric patients with acute lymphoblastic leukemia--implications for immunotherapy

Clin Cancer Res. 2013 Sep 15;19(18):5079-91. doi: 10.1158/1078-0432.CCR-13-0955. Epub 2013 Jul 9.


Purpose: Although modern cure rates for childhood acute lymphoblastic leukemia (ALL) exceed 80%, the outlook remains poor in patients with high-risk disease and those who relapse, especially when allogeneic hematopoietic stem cell transplantation is not feasible. Strategies to improve outcome and prevent relapse are therefore required. Immunotherapy with antigen-specific T cells can have antileukemic activity without the toxicities seen with intensive chemotherapy, and therefore represents an attractive strategy to improve the outcome of high-risk patients with ALL. We explored the feasibility of generating tumor antigen-specific T cells ex vivo from the peripheral blood of 50 patients with ALL [26 National Cancer Institute (NCI) high-risk and 24 standard-risk] receiving maintenance therapy.

Experimental design: Peripheral blood mononuclear cells were stimulated with autologous dendritic cells pulsed with complete peptide libraries of WT1, Survivin, MAGE-A3, and PRAME, antigens frequently expressed on ALL blasts.

Results: T-cell lines were successfully expanded from all patients, despite low lymphocyte counts and irrespective of NCI risk group. Antigen-specificity was observed in more than 50% of patients after the initial stimulation and increased to more than 90% after three stimulations as assessed in IFN-γ-enzyme-linked immunospot (ELISpot) and (51)Cr-release assays. Moreover, tumor-specific responses were observed by reduction of autologous leukemia blasts in short- and long-term coculture experiments.

Conclusion: This study supports the use of immunotherapy with adoptively transferred autologous tumor antigen-specific T cells to prevent relapse and improve the prognosis of patients with high-risk ALL.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Antigen-Presenting Cells / immunology
  • Antigens, Neoplasm / immunology*
  • Cell Proliferation
  • Cell- and Tissue-Based Therapy*
  • Child
  • Colony-Forming Units Assay
  • Dendritic Cells / immunology*
  • Enzyme-Linked Immunosorbent Assay
  • Feasibility Studies
  • HLA-A2 Antigen / immunology
  • Humans
  • Immunophenotyping
  • Immunotherapy*
  • Inhibitor of Apoptosis Proteins / immunology
  • Interferon-gamma / metabolism
  • Leukocytes, Mononuclear / immunology*
  • Neoplasm Proteins / immunology
  • Peptide Fragments / immunology
  • Precursor Cell Lymphoblastic Leukemia-Lymphoma / immunology
  • Precursor Cell Lymphoblastic Leukemia-Lymphoma / therapy*
  • Prognosis
  • Survivin
  • T-Lymphocytes, Cytotoxic / immunology*
  • Tumor Cells, Cultured
  • WT1 Proteins / immunology


  • Antigens, Neoplasm
  • BIRC5 protein, human
  • HLA-A2 Antigen
  • Inhibitor of Apoptosis Proteins
  • MAGEA3 protein, human
  • Neoplasm Proteins
  • PRAME protein, human
  • Peptide Fragments
  • Survivin
  • WT1 Proteins
  • Interferon-gamma