Langerhans-type and monocyte-derived human dendritic cells have different susceptibilities to mRNA electroporation with distinct effects on maturation and activation: implications for immunogenicity in dendritic cell-based immunotherapy

Journal of Translational Medicine

James W Young³  Erin T St Angelo²  Milena Mennecozzi¹  Justin A Shyer²  Katherine B Pronschinske²  Emanuela Romano⁴  David J Chung³ 

[1] European Commission, Joint Research Centre, Ispra (VA) 21027, Italy;Memorial Sloan-Kettering Cancer Center, New York, NY, USA;Weill Cornell Medical College, New York, NY, USA;Centre Hospitalier Universitaire Vaudois, CHUV-BH06, Rue du Bugnon 46, 1011, Lausanne,Switzerland/Suisse

关键词: Immunotherapy;    Cancer;    Dendritic cells;    mRNA electroporation;   

Others  :  827226
DOI  :  10.1186/1479-5876-11-166

 received in 2012-12-19, accepted in 2013-06-27,  发布年份 2013

【 摘 要 】

Background

mRNA electroporation of dendritic cells (DCs) facilitates processing and presentation of multiple peptides derived from whole antigen, tailored to different HLA molecules. Clinical responses to electroporated moDC vaccines, however, have been suboptimal. Human Langerhans-type DCs (LCs) are the most potent conventional DC subtype for inducing CD8⁺ cytotoxic T lymphocytes (CTLs) in vitro. We recently demonstrated that Wilms’ tumor 1 (WT1) mRNA-electroporated LCs are superior to moDCs as stimulators of tumor antigen-specific CD8⁺ CTLs, even though they are comparable stimulators of allogeneic T cell proliferative responses. A detailed comparative evaluation of the effects of mRNA electroporation on LCs versus moDCs, however, is needed.

Methods

Immature and partially-matured human moDCs and LCs electroporated with mRNA were compared for transfection efficiency, phenotypic changes, viability, retention of transgene expression after cryopreservation, and immunogenicity. Student t test was used for each pairwise comparison. One-way analysis of variance was used for multiple group comparisons.

Results

Transfection efficiency after electroporation with enhanced green fluorescent protein (eGFP) mRNA was higher for immature than for partially-matured moDCs. In contrast, transfection efficiency was higher for partially-matured than for immature LCs, with the additional benefit that electroporation itself increased maturation and activation of CD83⁺HLA-DR^bright LCs but not moDCs. Electroporation did not impair final maturation and activation of either DC subtype, after which both mRNA-electroporated LCs and moDCs were functionally similar in stimulating allogeneic T cell proliferation, a standard assay of DC immunogenicity.

Conclusions

These findings support mRNA electroporation of DCs, and in particular LCs, as an effective non-viral method to stimulate specific, potent CD8⁺ CTL responses. The differences between LCs and moDCs regarding this form of antigen-loading have important implications for DC-based immunotherapies.

【 授权许可】

   
2013 Chung et al.; licensee BioMed Central Ltd.

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