Electrofusion generates diverse DC-tumour cell hybrids for cancer immunotherapy.
BACKGROUND
Hybrid cells generated from dendritic cells (DC) and tumour cells provide tumour-associated antigens (TAA) in a polyvalent mode. The present study was designed to investigate the hybrid cell generation by dendritic cells and different tumour cell lines to establish an electrofusion protocol with an optimal fusion setting.


METHODS
Hybrid cells from mature DC and tumour cells were generated by electrofusion. Fusion efficiency was determined by flow cytometry, as well as by light and fluorescence microscopy analyses.


RESULTS
The gradual electrofusion process constituted different human dendritic cell tumour cell hybrids of high diversity depending on electrical and non-electrical parameters. Factors influencing fusion frequency were determined by specific cell staining with mAbs, FACS analysis and trypan blue dye exclusion.


CONCLUSIONS
Increased fusion efficiency was associated with reduced viability. The protocol presented in this work might be helpful for future fusion studies as a prerequisite for comparable in vitro and human vaccination trials.
Weise
Jan B
JB
Department of Otorhinolaryngology, Head and Neck Surgery, University of Kiel, Germany. jweise@hno.uni-kiel.de
Hilpert
Felix
F
Heiser
Axel
A
Görögh
Tibor
T
Meyer
Jens E
JE
Weimer
Joerg
J
Krüger
Christoph
C
Rosenau
Andreas
A
Wieckhorst
Wilm
W
Arnold
Norbert
N
Pfisterer
Jacobus
J
Kabelitz
Dietrich
D
Maune
Steffen
S
D000951:Antigens, Neoplasm / Q000276:immunology; D002294:Carcinoma, Squamous Cell / Q000276:immunology* / Q000473:pathology*; D002459:Cell Fusion / Q000379:methods*; D003713:Dendritic Cells / Q000166:cytology* / Q000276:immunology*; D005434:Flow Cytometry; D006684:HLA-DR Antigens / Q000276:immunology; D006801:Humans; D006822:Hybrid Cells / Q000166:cytology / Q000276:immunology; D016219:Immunotherapy, Adoptive / Q000379:methods*; D007822:Laryngeal Neoplasms / Q000276:immunology* / Q000473:pathology*
D000951:Antigens, Neoplasm; D006684:HLA-DR Antigens