title: Gene Expression Profile of Dendritic Cell-Tumor Cell Hybrids Determined by Microarrays and Its Implications for Cancer Immunotherapy.
abstract: BACKGROUND
  Dendritic cell- (DC-) tumor fusion cells stimulate effective in vivo antitumor responses. However, therapeutic approaches are dependent upon the coadministration of exogenous 3rd signals. The purpose of this study was to determine the mechanisms for inadequate 3rd signaling by electrofused DC-tumor cell hybrids.
  
  
  METHODS
  Murine melanoma cells were fused with DCs derived from C57BL/6 mice. Quantitative real-time PCR (qPCR) was used to determine relative changes in Th (T helper) 1 and Th2 cytokine gene expression. In addition, changes in gene expression of fusion cells were determined by microarray. Last, cytokine secretion by fusion cells upon inhibition of signaling pathways was analyzed by ELISA.
  
  
  RESULTS
  qPCR analyses revealed that fusion cells exhibited a downregulation of Th1 associated cytokines IL-12 and IL-15 and an upregulation of the Th2 cytokine IL-4. Microarray studies further showed that the expression of chemokines, costimulatory molecules, and matrix-metalloproteinases was deregulated in fusion cells. Lastly, inhibitor studies demonstrate that inhibition of the PI3K/Akt/mTOR signaling pathway could restore the secretion of bioactive IL-12p70 by fusion cells.
  
  
  CONCLUSIONS
  Our results suggest that combining fusion cell-based vaccination with administration of inhibitors of the PI3K/Akt/mTOR signaling pathway may enhance antitumor responses in patients.
authors.affiliation: Department of Surgery, Duke University Medical Center, Durham, NC 27710, USA.
authors.forename: Jens
authors.initials: J
authors.lastname: Dannull
authors.affiliation: Department of Surgery, Division of Otolaryngology-Head and Neck Surgery, Duke University Medical Center, Durham, NC 27710, USA.
authors.forename: Chunrui
authors.initials: C
authors.lastname: Tan
authors.affiliation: Duke University, Durham, NC 27710, USA.
authors.forename: Christine
authors.initials: C
authors.lastname: Farrell
authors.affiliation: Duke University School of Medicine, Durham, NC 27710, USA.
authors.forename: Cynthia
authors.initials: C
authors.lastname: Wang
authors.affiliation: Department of Surgery, Duke University Medical Center, Durham, NC 27710, USA ; Division of Experimental Medicine, Merck, Rahway, NJ 07065, USA.
authors.forename: Scott
authors.initials: S
authors.lastname: Pruitt
authors.affiliation: Department of Surgery, Duke University Medical Center, Durham, NC 27710, USA.
authors.forename: Smita K
authors.initials: SK
authors.lastname: Nair
authors.affiliation: Department of Surgery, Division of Otolaryngology-Head and Neck Surgery, Duke University Medical Center, Durham, NC 27710, USA ; Duke University School of Medicine, Durham, NC 27710, USA ; Durham VA Medical Center, Section of Otolaryngology-Head and Neck Surgery, Durham, NC 27705, USA.
authors.forename: Walter T
authors.initials: WT
authors.lastname: Lee
mesh_terms: D000818:Animals; D002459:Cell Fusion; D045744:Cell Line, Tumor; D018925:Chemokines / Q000235:genetics / Q000378:metabolism; D016207:Cytokines / Q000235:genetics / Q000378:metabolism; D003713:Dendritic Cells / Q000276:immunology* / Q000378:metabolism*; D005260:Female; D020869:Gene Expression Profiling; D006822:Hybrid Cells / Q000378:metabolism*; D007167:Immunotherapy / Q000379:methods; D020782:Matrix Metalloproteinases / Q000235:genetics / Q000378:metabolism; D008546:Melanoma, Experimental; D051379:Mice; D009369:Neoplasms / Q000235:genetics* / Q000276:immunology* / Q000628:therapy; D019707:Receptors, Chemokine / Q000235:genetics / Q000378:metabolism; D018121:Receptors, Cytokine / Q000235:genetics / Q000378:metabolism; D015398:Signal Transduction; D059467:Transcriptome
chemical_list: D018925:Chemokines; D016207:Cytokines; D019707:Receptors, Chemokine; D018121:Receptors, Cytokine; D020782:Matrix Metalloproteinases