Dendritic cell-based genetic immunization in mice with a recombinant adenovirus encoding murine TRP2 induces effective anti-melanoma immunity
✍ Scribed by Thomas Tüting; Julia Steitz; Jürgen Brück; Andrea Gambotto; Kerstin Steinbrink; Albert B. DeLeo; Paul Robbins; Jürgen Knop; Alexander H. Enk
- Book ID
- 101300247
- Publisher
- John Wiley and Sons
- Year
- 1999
- Tongue
- English
- Weight
- 175 KB
- Volume
- 1
- Category
- Article
- ISSN
- 1099-498X
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✦ Synopsis
Background
The induction of cellular immune responses to melanocyte-speci®c enzymes such as the tyrosinase family of proteins is the goal of various clinical studies for the immunotherapy of melanoma. Tyrosinase-related protein-2 (TRP2) is an attractive model antigen for preclinical studies in C57BL/6 mice because it is naturally expressed by the murine B16 melanoma and can be recognized by self-reactive cytolytic T lymphocytes (CTL). Here we describe efforts to develop genetic immunization with dendritic cells (DC) for the immunotherapy of melanoma in this clinically relevant system.
Methods
Recombinant adenoviruses encoding green ¯uorescent protein (Ad-EGFP) and murine TRP2 (Ad-mTRP2) were constructed using Cre-loxPmediated recombination. DC were generated in vitro from precursors in bone marrow and transduced with Ad-EGFP or Ad-mTRP2. Mice were immunized by direct injection of adenovirus or by injection of Ad-transduced DC. Induction of tumor immunity was assessed by intravenous challenge with B16 melanoma cells and enumeration of experimentally induced lung metastases.
Results
Flowcytometric analysis of DC transduced with Ad-EGFP demonstrated endogenous ¯uorescence due to cytoplasmatic expression of EGFP in 30±60% of cells. Ad-EGFP-transduced DC simultaneously displayed the DC-speci®c marker NLDC145 and high levels of MHC and costimulatory molecules on their cell surface. Transduction of DC with Ad-mTRP2 resulted in strong intracellular expression of TRP2 which could be readily detected by immunostaining. Importantly, immunization of mice with cultured Ad-mTRP2-transduced DC completely prevented the development of lung metastases following an intravenous challenge with B16 melanoma cells. This striking protective effect was observed with both the intravenous and the subcutaneous route of DC immunization. In vivo depletion of T-cell subsets suggested that the protective effect of an immunization with Ad-mTRP2transduced DC involved both CD8 + and CD4 + T-cells.
Conclusions
Our results demonstrate that DC-based genetic immunization of mice with TRP2, a clinically relevant melanocyte-speci®c self-antigen, induces effective cellular immunity and prevents metastatic growth of B16 melanoma cells in vivo.