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Gene Ther. 2015 Sep;22(9):707-20. doi: 10.1038/gt.2015.43. Epub 2015 May 28.

Lentivirus-induced 'Smart' dendritic cells: Pharmacodynamics and GMP-compliant production for immunotherapy against TRP2-positive melanoma.

Author information

1
Department of Hematology, Hemostasis, Oncology and Stem Cell Transplantation, Hannover Medical School, Hannover, Germany.
2
Department of Hematological Medicine, Cell and Gene Therapy at King's, The Rayne Institute, King's College London, London, UK.
3
Department of Dermatology and Allergy, Skin Cancer Center Hannover, Hannover Medical School, Hannover, Germany.
4
EUFETS GmbH, Idar-Oberstein, Heidelberg, Germany.
5
Division of Translational Oncology, National Center for Tumor Diseases, Heidelberg, Germany.
6
Department of Experimental Hematology, Hannover, Germany.
7
John Wayne Cancer Institute, Santa Monica, CA, USA.
8
Department of Transfusion Medicine, Hannover Medical School, Hannover, Germany.
9
Institute for Cell Therapeutics and GMP core facility IFB-Tx, Hannover Medical School, Hannover, Germany.

Abstract

Monocyte-derived conventional dendritic cells (ConvDCs) loaded with melanoma antigens showed modest responses in clinical trials. Efficacy studies were hampered by difficulties in ConvDC manufacturing and low potency. Overcoming these issues, we demonstrated higher potency of lentiviral vector (LV)-programmed DCs. Monocytes were directly induced to self-differentiate into DCs (SmartDC-TRP2) upon transduction with a tricistronic LV encoding for cytokines (granulocyte macrophage colony stimulating factor (GM-CSF) and interleukin-4 (IL-4)) and a melanoma antigen (tyrosinase-related protein 2 (TRP2)). Here, SmartDC-TRP2 generated with monocytes from five advanced melanoma patients were tested in autologous DC:T cell stimulation assays, validating the activation of functional TRP2-specific cytotoxic T lymphocytes (CTLs) for all patients. We described methods compliant to good manufacturing practices (GMP) to produce LV and SmartDC-TRP2. Feasibility of monocyte transduction in a bag system and cryopreservation following a 24-h standard operating procedure were achieved. After thawing, 50% of the initial monocyte input was recovered and SmartDC-TRP2 self-differentiated in vitro, showing uniform expression of DC markers, detectable LV copies and a polyclonal LV integration pattern not biased to oncogenic loci. GMP-grade SmartDC-TRP2 expanded TRP2-specific autologous CTLs in vitro. These results demonstrated a simpler GMP-compliant method of manufacturing an effective individualized DC vaccine. Such DC vaccine, when in combination with checkpoint inhibition therapies, might provide higher specificity against melanoma.

PMID:
25965393
PMCID:
PMC4561294
DOI:
10.1038/gt.2015.43
[Indexed for MEDLINE]
Free PMC Article

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