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

Authors :
Eliana Ruggiero
Klaus Kuehlcke
Bala Sai Sundarasetty
Laura Gerasch
Farzin Farzaneh
Rainer Blasczyk
F Schenck
Constanca Figueiredo
D S B Hoon
Michael Rothe
David Darling
S. Naundorf
Renata Stripecke
C von Kalle
Lucas Chan
Manfred G. Schmidt
Ralf Gutzmer
Giulia Giunti
Ulrike Koehl
Source :
Chan, L 2015, ' Lentivirus-induced 'Smart' dendritic cells : Pharmacodynamics and GMP-compliant production for immunotherapy against TRP2 positive melanoma ', Gene Therapy, vol. 22, pp. 707-720 . https://doi.org/10.1038/gt.2015.43, Gene Therapy
Publication Year :
2015
Publisher :
Springer Science and Business Media LLC, 2015.

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 (SmartDCTRP2) 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 selfdifferentiated 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 asimpler 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.

Details

ISSN :
14765462 and 09697128
Volume :
22
Database :
OpenAIRE
Journal :
Gene Therapy
Accession number :
edsair.doi.dedup.....bb8da118b9831cb41d51e9a6cd777e41