title: Characteristics of hybrid cells obtained by dendritic cell/tumour cell fusion in a T-47D breast cancer cell line model indicate their potential as anti-tumour vaccines.
abstract: Many strategies have been proposed to circumvent cancer development or prevent its growth. One of the promising strategies is to direct the immune response toward tumour antigens. This can be achieved by loading dendritic cells, the most potent antigen presenting cells, with tumour antigens. Fusion of dendritic cells (DC) with tumour cells is an attractive way to load the DC with all tumour antigens regardless of their immunogenicity status and the fact that they have, or not, been identified. The aim of our study was to characterise the immunophenotype of fused cells, monitor the evolution of the fusion interface and the distribution of surface antigens over time and assess for their maturation status and functionality in vitro. We used polyethylene glycol to fuse DC with Her2/neu positive breast cancer cell line T-47D. We demonstrate that false positive events accounted in flow cytometry can be identified using confocal microscopy to avoid an overestimation of fusion efficiency and to distinguish clearly hybrid cells from aggregated or phagocytosed cells. We used imaging means to demonstrate the conservation of presentation molecules (MHC II, CD1a), co-stimulatory molecules (CD40, CD80, CD86), as well as tumour antigens (Her2/neu, cytokeratins) in optimised conditions. Fused cells were only recognisable for 48 h as assessed by membrane staining and membranous antigen distribution. Fusion was necessary for their maturation to be accompanied by functional activity such as secretion of cytokines and perforin. These results suggest that hybrid cells generated by the fusion of DC and tumour cells can be easily identified and characterised using imaging techniques, and that, regarding functionality and cytokine secretion, they appear to be good candidates for anti-tumour therapies namely in breast cancer.
authors.affiliation: EA3499, Université Paris 6, Laboratoire d'Histologie Biologie Tumorale, Hôpital Tenon, AP-HP, Paris, France.
authors.forename: Karine
authors.initials: K
authors.lastname: Serhal
authors.forename: Claude
authors.initials: C
authors.lastname: Baillou
authors.forename: Nicolae
authors.initials: N
authors.lastname: Ghinea
authors.forename: Philippe
authors.initials: P
authors.lastname: Fontanges
authors.forename: Franck P
authors.initials: FP
authors.lastname: Dupuy
authors.forename: François M
authors.initials: FM
authors.lastname: Lemoine
authors.forename: Roger
authors.initials: R
authors.lastname: Lacave
mesh_terms: D001943:Breast Neoplasms / Q000276:immunology / Q000473:pathology / Q000628:therapy*; D019496:Cancer Vaccines / Q000627:therapeutic use*; D002459:Cell Fusion; D045744:Cell Line, Tumor; D003713:Dendritic Cells / Q000502:physiology*; D005260:Female; D005434:Flow Cytometry; D006684:HLA-DR Antigens / Q000032:analysis; D006801:Humans; D006822:Hybrid Cells / Q000502:physiology*; D016753:Interleukin-10 / Q000096:biosynthesis; D018719:Erb-b2 Receptor Tyrosine Kinases / Q000032:analysis
chemical_list: D019496:Cancer Vaccines; D006684:HLA-DR Antigens; D016753:Interleukin-10; D018719:Erb-b2 Receptor Tyrosine Kinases