Generation of dendritic cell-tumor cell hybrids by electrofusion for clinical vaccine application

被引:41
作者
Trevor, KT
Cover, C
Ruiz, YW
Akporiaye, ET
Hersh, EM
Landais, D
Taylor, RR
King, AD
Walters, RE
机构
[1] Arizona Canc Ctr, Tucson, AZ 85748 USA
[2] IDM, Paris, France
[3] Cyto Pulse Sci, Columbia, MD USA
关键词
cancer; dendritic cells; electrofusion; hybrids; immunotherapy;
D O I
10.1007/s00262-004-0512-1
中图分类号
R73 [肿瘤学];
学科分类号
100214 ;
摘要
Vaccination with hybrids comprising fused dendritic cells (DCs) and tumor cells is a novel cancer immunotherapy approach designed to combine tumor antigenicity with the antigen-presenting and immune-stimulatory capacities of DCs. For clinical purposes, we have incorporated a large-scale process for the generation of clinical-grade DCs together with novel electrofusion technology. The electrofusion system provides for ease and standardization of method, efficient DC-tumor cell hybrid formation, and large-quantity production of hybrids in a high-volume (6-ml) electrofusion chamber. In addition, we have evaluated DC electrofusion with a variety of allogeneic human tumor cell lines with the rationale that these tumor cell partners would prove a ready, suitable source for the generation of DC-tumor cell hybrid vaccines. The DC production process can generate 6x10(8) to 2x10(9) DCs from a single leukapheresis product (similar to180 ml). As determined by FACS analysis, electrofusion of 6x10(7) total cells (1:1 ratio of DC and tumor cells) resulted in a consistent average of 8-10% DC-tumor cell hybrids, irrespective of the tumor type used. Hybrids were retained in the population for 48 h postfusion and following freezing and thawing. Upon pre-irradiation of the tumor cell partner for vaccine purposes, the overall fusion efficiency was not altered at doses up to 200 Gy. Evaluation of DC-tumor cell hybrid populations for their ability to stimulate T-cell responses demonstrated that electrofused populations are superior to mixed populations of DCs and tumor cells in generating a primary T-cell response, as indicated by IFN-gamma release. Moreover, hybrids comprising HLA-A(*)0201 DCs and allogeneic melanoma tumor cells (Colo 829 cell line) stimulated IFN-gamma secretion by antigen-specific CD8(+) T cells, which are restricted for recognition of a melanoma gp100 peptide antigen (gp100(209-217)) within the context of the DC HLA haplotype. Maturation of the DC-Colo 829 cell hybrid population served to further improve this T-cell gp100-specific response. Overall, our results are promising for the large-scale generation of electrofused hybrids comprising DCs and allogeneic tumor cells, that may prove useful in human vaccine trials.
引用
收藏
页码:705 / 714
页数:10
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