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

Katrina T. Trevor, Cathleen Cover, Yvette W. Ruiz, Emmanuel T. Akporiaye, Evan M Hersh, Didier Landais, Rachel R. Taylor, Alan D. King, Richard E. Walters

Research output: Contribution to journalArticle

49 Citations (Scopus)

Abstract

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 immunestimulatory 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 6×108 to 2×109 DCs from a single leukapheresis product (∼180 ml). As determined by FACS analysis, electrofusion of 6×107 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-γ release. Moreover, hybrids comprising HLA-A*0201 DCs and allogeneic melanoma tumor cells (Colo 829 cell line) stimulated IFN-γ secretion by antigen-specific CD8+ T cells, which are restricted for recognition of a melanoma gp100 peptide antigen (gp100209-217) 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.

Original languageEnglish (US)
Pages (from-to)705-714
Number of pages10
JournalCancer Immunology, Immunotherapy
Volume53
Issue number8
StatePublished - Aug 2004

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Hybrid Cells
Dendritic Cells
Vaccines
Neoplasms
T-Lymphocytes
Population
gp100 Melanoma Antigen
CD8 Antigens
Leukapheresis
Tumor Cell Line
Immunotherapy
Haplotypes

Keywords

  • Cancer
  • Dendritic cells
  • Electrofusion
  • Hybrids
  • Immunotherapy

ASJC Scopus subject areas

  • Cancer Research
  • Immunology
  • Oncology

Cite this

Trevor, K. T., Cover, C., Ruiz, Y. W., Akporiaye, E. T., Hersh, E. M., Landais, D., ... Walters, R. E. (2004). Generation of dendritic cell-tumor cell hybrids by electrofusion for clinical vaccine application. Cancer Immunology, Immunotherapy, 53(8), 705-714.

Generation of dendritic cell-tumor cell hybrids by electrofusion for clinical vaccine application. / Trevor, Katrina T.; Cover, Cathleen; Ruiz, Yvette W.; Akporiaye, Emmanuel T.; Hersh, Evan M; Landais, Didier; Taylor, Rachel R.; King, Alan D.; Walters, Richard E.

In: Cancer Immunology, Immunotherapy, Vol. 53, No. 8, 08.2004, p. 705-714.

Research output: Contribution to journalArticle

Trevor, KT, Cover, C, Ruiz, YW, Akporiaye, ET, Hersh, EM, Landais, D, Taylor, RR, King, AD & Walters, RE 2004, 'Generation of dendritic cell-tumor cell hybrids by electrofusion for clinical vaccine application', Cancer Immunology, Immunotherapy, vol. 53, no. 8, pp. 705-714.
Trevor, Katrina T. ; Cover, Cathleen ; Ruiz, Yvette W. ; Akporiaye, Emmanuel T. ; Hersh, Evan M ; Landais, Didier ; Taylor, Rachel R. ; King, Alan D. ; Walters, Richard E. / Generation of dendritic cell-tumor cell hybrids by electrofusion for clinical vaccine application. In: Cancer Immunology, Immunotherapy. 2004 ; Vol. 53, No. 8. pp. 705-714.
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