Carcinoembryonic antigen (CEA) presentation and specific T cell-priming by human dendritic cells transfected with CEA-mRNA.
The feasibility of dendritic cells (DC) for cancer immunotherapy after transfection by electroporation with mRNA encoding the human carcinoembryonic antigen (CEA) was investigated. Both, total RNA from the CEA(+) colon cancer cell line SW480 and mRNA transcribed in vitro from cDNA3.1-plasmids (pcDNA3.1+/-HisC) with a CEA-insert (ivt-CEA-mRNA, ivt-CEA/HisC-mRNA) were used. Labelled ivt-CEA-mRNA was detectable in DC by light and electron microscopy and by fluorescence-activated cell-sorting (FACS) even 15 min after electroporation. Four hours after transfection with ivt-CEA/HisC-mRNA, we detected specific expression of CEA and the histidine-tag by immunofluorescence microscopy and by FACS. CEA-specific T lymphocytes were successfully primed by transfected DC and were able to lyse CEA-expressing target cells, even from the CEA-expressing human colon adenocarcinoma cell line SW480. Thus, DC transfected by electroporation with CEA-mRNA are valuable tools for the immunotherapy of CEA(+) tumour entities.
Eppler
E
E
Institute of Anatomy, Division of Cell Biology, University of Zurich, Zurich, Switzerland. eppler@anatom.unizh.ch
Hörig
H
H
Kaufman
H L
HL
Groscurth
P
P
Filgueira
L
L
D017951:Antigen Presentation; D015496:CD4-Positive T-Lymphocytes / Q000276:immunology; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology; D002272:Carcinoembryonic Antigen / Q000276:immunology*; D003110:Colonic Neoplasms / Q000628:therapy*; D003713:Dendritic Cells / Q000276:immunology*; D018274:Electroporation / Q000379:methods; D006801:Humans; D016233:Immunotherapy, Active; D012333:RNA, Messenger / Q000276:immunology; D013601:T-Lymphocytes / Q000276:immunology*; D014162:Transfection / Q000379:methods*
D002272:Carcinoembryonic Antigen; D012333:RNA, Messenger