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Cancer Immunology, Immunotherapy

, Volume 55, Issue 12, pp 1480–1490 | Cite as

In vitro priming of tumor-specific cytotoxic T lymphocytes using allogeneic dendritic cells derived from the human MUTZ-3 cell line

  • Saskia J. A. M. Santegoets
  • Marco W. J. Schreurs
  • Allan J. Masterson
  • Ying Poi Liu
  • Steffen Goletz
  • Hans Baumeister
  • Esther W. M. Kueter
  • Sinéad M. Lougheed
  • Alfons J. M. van den Eertwegh
  • Rik J. Scheper
  • Erik Hooijberg
  • Tanja D. de Gruijl
Original Article

Abstract

The adoptive transfer of in vitro-induced and expanded tumor-specific cytotoxic T lymphocytes (CTL) presents a promising immunotherapeutic approach for the treatment of cancer. The in vitro induction of tumor-reactive CTL requires repeated stimulation of CTL precursors with dendritic cells (DC). To circumvent problems like scarcity of blood DC precursors and donor variability, it would be attractive to use DC from a non-autologous, unlimited source. DCs derived from the human acute myeloid leukemia (AML) cell line MUTZ-3 are attractive candidates since these DCs closely resemble monocyte-derived DC (MoDC) in terms of phenotype and T cell stimulatory capacity. Here we demonstrate that functional CTL clones could be generated against multiple tumor-associated antigens, i.e., human telomerase reverse transcriptase (hTERT), ErbB3-binding protein-1 (Ebp1), carcinoembryonic antigen (CEA) and Her-2/neu, by stimulating CD8β+ CTL precursors with peptide-loaded allogeneic, HLA-A2-matched MUTZ-3-derived DC. A consistent induction capacity, as determined by MHC tetramer-binding, was found in multiple donors and comparable to autologous peptide-loaded MoDC. Functional characterization at the clonal level revealed the priming of CTL that recognized endogenously processed epitopes on tumor cell lines in an HLA-A2-restricted fashion. Our data indicate that MUTZ-3-derived DC can be used as stimulator cells for in vitro priming and expansion of functional TAA-specific effector CTL. MUTZ-3-derived DCs thus represent a ready and standardized source of allogeneic DC to generate CTL for therapeutic adoptive transfer strategies.

Keywords

Human dendritic cell line Tumor-associated antigens Immunization Immunotherapy Adoptive T cell transfer 

Abbreviations

APC

Antigen-presenting cell

CEA

Carcinoembryonic antigen

CTL

Cytotoxic T lymphocytes

DC

Dendritic cell

GFP

Green fluorescent protein

hTERT

Human telomerase reverse transcriptase

MoDC

Monocyte-derived dendritic cell

ΔNGFR

Truncated form of nerve growth factor receptor

PBMC

Peripheral blood mononuclear cell

PHA

Phytohemagglutin

TIL

Tumor-infiltrating lymphocyte

Tm

Tetramer

Notes

Acknowledgments

The authors wish to thank NEMOD Biotherapeutics for their financial support and the Maurits & Anna de Kock Foundation for financial support in the purchase of an HPLC.

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Copyright information

© Springer-Verlag 2006

Authors and Affiliations

  • Saskia J. A. M. Santegoets
    • 1
  • Marco W. J. Schreurs
    • 1
  • Allan J. Masterson
    • 2
  • Ying Poi Liu
    • 1
  • Steffen Goletz
    • 3
    • 4
  • Hans Baumeister
    • 4
  • Esther W. M. Kueter
    • 1
  • Sinéad M. Lougheed
    • 2
  • Alfons J. M. van den Eertwegh
    • 2
  • Rik J. Scheper
    • 1
    • 5
  • Erik Hooijberg
    • 1
  • Tanja D. de Gruijl
    • 2
  1. 1.Departments of PathologyVU University Medical CenterAmsterdamThe Netherlands
  2. 2.Medical OncologyVU University Medical CenterAmsterdamThe Netherlands
  3. 3.NEMOD BiotherapeuticsGmbH & Co. KGBerlinGermany
  4. 4.GLYCOTOPE GmbHBerlinGermany
  5. 5.AmsterdamThe Netherlands

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