title: In vitro generation of cytotoxic and regulatory T cells by fusions of human dendritic cells and hepatocellular carcinoma cells.
abstract: BACKGROUND
  Human hepatocellular carcinoma (HCC) cells express WT1 and/or carcinoembryonic antigen (CEA) as potential targets for the induction of antitumor immunity. In this study, generation of cytotoxic T lymphocytes (CTL) and regulatory T cells (Treg) by fusions of dendritic cells (DCs) and HCC cells was examined.
  
  
  METHODS
  HCC cells were fused to DCs either from healthy donors or the HCC patient and investigated whether supernatants derived from the HCC cell culture (HCCsp) influenced on the function of DCs/HCC fusion cells (FCs) and generation of CTL and Treg.
  
  
  RESULTS
  FCs coexpressed the HCC cells-derived WT1 and CEA antigens and DCs-derived MHC class II and costimulatory molecules. In addition, FCs were effective in activating CD4+ and CD8+ T cells able to produce IFN-gamma and inducing cytolysis of autologous tumor or semiallogeneic targets by a MHC class I-restricted mechanism. However, HCCsp induced functional impairment of DCs as demonstrated by the down-regulation of MHC class I and II, CD80, CD86, and CD83 molecules. Moreover, the HCCsp-exposed DCs failed to undergo full maturation upon stimulation with the Toll-like receptor 4 agonist penicillin-inactivated Streptococcus pyogenes. Interestingly, fusions of immature DCs generated in the presence of HCCsp and allogeneic HCC cells promoted the generation of CD4+ CD25high Foxp3+ Treg and inhibited CTL induction in the presence of HCCsp. Importantly, up-regulation of MHC class II, CD80, and CD83 on DCs was observed in the patient with advanced HCC after vaccination with autologous FCs. In addition, the FCs induced WT1- and CEA-specific CTL that were able to produce high levels of IFN-gamma.
  
  
  CONCLUSIONS
  The current study is one of the first demonstrating the induction of antigen-specific CTL and the generation of Treg by fusions of DCs and HCC cells. The local tumor-related factors may favor the generation of Treg through the inhibition of DCs maturation; however, fusion cell vaccination results in recovery of the DCs function and induction of antigen-specific CTL responses in vitro. The present study may shed new light about the mechanisms responsible for the generation of CTL and Treg by FCs.
authors.affiliation: Division of Gastroenterology and Hepatology, Department of Internal Medicine, The Jikei University School of Medicine, Tokyo, Japan. shigeo_koido@jikei.ac.jp
authors.forename: Shigeo
authors.initials: S
authors.lastname: Koido
authors.forename: Sadamu
authors.initials: S
authors.lastname: Homma
authors.forename: Eiichi
authors.initials: E
authors.lastname: Hara
authors.forename: Makoto
authors.initials: M
authors.lastname: Mitsunaga
authors.forename: Yoshihisa
authors.initials: Y
authors.lastname: Namiki
authors.forename: Akitaka
authors.initials: A
authors.lastname: Takahara
authors.forename: Eijiro
authors.initials: E
authors.lastname: Nagasaki
authors.forename: Hideo
authors.initials: H
authors.lastname: Komita
authors.forename: Yukiko
authors.initials: Y
authors.lastname: Sagawa
authors.forename: Toshifumi
authors.initials: T
authors.lastname: Ohkusa
authors.forename: Kiyotaka
authors.initials: K
authors.lastname: Fujise
authors.forename: Jianlin
authors.initials: J
authors.lastname: Gong
authors.forename: Hisao
authors.initials: H
authors.lastname: Tajiri
mesh_terms: D015704:CD4 Antigens / Q000378:metabolism; D002272:Carcinoembryonic Antigen / Q000378:metabolism; D006528:Carcinoma, Hepatocellular / Q000276:immunology / Q000473:pathology*; D002454:Cell Differentiation; D002459:Cell Fusion / Q000379:methods*; D045744:Cell Line, Tumor; D003713:Dendritic Cells / Q000166:cytology* / Q000276:immunology; D051858:Forkhead Transcription Factors / Q000378:metabolism; D006801:Humans; D053645:Interleukin-2 Receptor alpha Subunit / Q000378:metabolism; D008113:Liver Neoplasms / Q000276:immunology / Q000473:pathology*; D008297:Male; D008875:Middle Aged; D010641:Phenotype; D013347:Subcellular Fractions / Q000378:metabolism; D013602:T-Lymphocytes, Cytotoxic / Q000166:cytology* / Q000276:immunology; D050378:T-Lymphocytes, Regulatory / Q000166:cytology* / Q000276:immunology; D014611:Vaccination; D025721:WT1 Proteins / Q000378:metabolism
chemical_list: D015704:CD4 Antigens; D002272:Carcinoembryonic Antigen; C418974:FOXP3 protein, human; D051858:Forkhead Transcription Factors; D053645:Interleukin-2 Receptor alpha Subunit; D025721:WT1 Proteins
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