Human dendritic cells transfected with either RNA or DNA encoding influenza matrix protein M1 differ in their ability to stimulate cytotoxic T lymphocytes.
The use of tumor antigen loaded dendritic cells (DC) is one of the most promising approaches to induce a tumor specific immune response in vivo. Several strategies have been designed to load DC with tumor antigens. In this study, we investigated the delivery of in vitro transcribed RNA and plasmid DNA into monocyte-derived, ie non-proliferating human DC, using several nonviral transfection methods including electroporation and lipofection. Green fluorescent protein (GFP) was used as a reporter gene and influenza matrix protein 1 (M1) as a model antigen for HLA class I restricted antigen presentation. Using electroporation in combination with DNA or with RNA, up to 11% of DC were GFP-positive. Using liposomes as a vehicle for DNA transport up to 10% of the DC were GFP-positive. A significant increase in transfection efficacy, of up to 20%, was observed when GFP RNA was used in combination with liposomes. Importantly, the RNA transfected DC retained their typical morphological and immunophenotypical characteristics. In addition, DC transfected with M1 RNA were able to stimulate autologous peripheral M1-specific memory cytotoxic T lymphocytes (CTL), as well as M1-specific CTL clones. Furthermore, comparison of DNA-transfected DC with RNA-transfected DC revealed the latter to be far better stimulators of antigen-specific T cells. This RNA transfection technique consequently represents a very promising tool for future immunotherapy strategies.
Strobel
I
I
Department of Dermatology, University of Erlangen, Germany.
Berchtold
S
S
Götze
A
A
Schulze
U
U
Schuler
G
G
Steinkasserer
A
A
D000956:Antigens, Viral / Q000235:genetics*; D002999:Clone Cells; D004247:DNA / Q000008:administration & dosage; D003713:Dendritic Cells / Q000276:immunology*; D018274:Electroporation; D015870:Gene Expression; D015316:Genetic Therapy / Q000379:methods*; D049452:Green Fluorescent Proteins; D006801:Humans; D007156:Immunologic Memory; D007167:Immunotherapy; D008081:Liposomes; D008164:Luminescent Proteins / Q000235:genetics; D008213:Lymphocyte Activation; D012313:RNA / Q000008:administration & dosage; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology*; D014162:Transfection / Q000379:methods*; D014763:Viral Matrix Proteins / Q000235:genetics*
D000956:Antigens, Viral; D008081:Liposomes; D008164:Luminescent Proteins; C031540:M-protein, influenza virus; C488936:M1 protein, Influenza A virus; D014763:Viral Matrix Proteins; D049452:Green Fluorescent Proteins; D012313:RNA; D004247:DNA