title: Exposure to the electrofusion process can increase the immunogenicity of human cells.
abstract: The cellular products obtained following electrofusion (EF) of dendritic cells (DC) and tumour cells have shown promise as cancer vaccines. The immunogenicity of these preparations has been attributed to the presence of small numbers of DC-tumour hybrids and the contribution of the non-hybrid tumour cells present has received little attention. In this report, we investigated the effect of the EF process on the immunogenicity of allogeneic human cells, in particular the colorectal cell line, SW620. EF conditions were optimised to yield the maximum number of DC-SW620 hybrids co-expressing tumour associated antigen (TAA) and DC associated antigens. Exposure of SW620 to EF induced significant increases (P < 0.05) in apoptosis and necrosis. Pre-exposure of SW620 to the EF buffer alone [0.3 M glucose, 0.1 mM Ca(CH3COO)2 and 0.5 mM Mg(CH3COO)(2)] resulted in significant increases in TAA uptake by DC during co-culture (P < 0.05). DC phenotype was, however, not altered by exposure to EF treated tumour cells. In co-cultures of PBMC responders with SW620, the levels of IFNgamma release and cytotoxic activity were significantly increased (P < 0.05) by pre-exposure of the SW620 to EF. Pre-exposure of allogeneic non-T cells, the colorectal cell line Lovo and a breast cancer cell line (MCF7) to EF also significantly (P < 0.05) increased the levels of IFNgamma release by responding PBMC. These results demonstrate that the EF process itself can increase the immunogenicity of at least some human cell types independently of hybrid formation. These findings suggest that EF protocols should be evaluated with regard to the possibility that DC-tumour hybrids may not contribute all, or even most, of the immunostimulatory capacity present in preparations of EF treated cells.
authors.affiliation: Haematology Research Group, Christchurch School Of Medicine, PO Box 4345, Christchurch, New Zealand. barry.hock@chmeds.ac.nz
authors.forename: Barry D
authors.initials: BD
authors.lastname: Hock
authors.forename: Georgina
authors.initials: G
authors.lastname: Roberts
authors.forename: Judith L
authors.initials: JL
authors.lastname: McKenzie
authors.forename: Prachee
authors.initials: P
authors.lastname: Gokhale
authors.forename: Nina
authors.initials: N
authors.lastname: Salm
authors.forename: Alexander D
authors.initials: AD
authors.lastname: McLellan
authors.forename: Nigel W
authors.initials: NW
authors.lastname: Patton
authors.forename: Justin A
authors.initials: JA
authors.lastname: Roake
mesh_terms: D000938:Antigen-Presenting Cells / Q000276:immunology*; D000951:Antigens, Neoplasm / Q000276:immunology; D017209:Apoptosis / Q000276:immunology; D001943:Breast Neoplasms / Q000276:immunology* / Q000473:pathology; D019496:Cancer Vaccines / Q000276:immunology*; D002459:Cell Fusion; D002470:Cell Survival / Q000276:immunology; D003110:Colonic Neoplasms / Q000276:immunology* / Q000473:pathology; D003713:Dendritic Cells / Q000276:immunology*; D018274:Electroporation / Q000379:methods; D006801:Humans; D007371:Interferon-gamma / Q000378:metabolism; D009336:Necrosis; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology; D014407:Tumor Cells, Cultured / Q000276:immunology / Q000378:metabolism
chemical_list: D000951:Antigens, Neoplasm; D019496:Cancer Vaccines; D007371:Interferon-gamma