In vitro generation of cytotoxic and regulatory T cells by fusions of human dendritic cells and hepatocellular carcinoma cells.
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.
Koido
Shigeo
S
Division of Gastroenterology and Hepatology, Department of Internal Medicine, The Jikei University School of Medicine, Tokyo, Japan. shigeo_koido@jikei.ac.jp
Homma
Sadamu
S
Hara
Eiichi
E
Mitsunaga
Makoto
M
Namiki
Yoshihisa
Y
Takahara
Akitaka
A
Nagasaki
Eijiro
E
Komita
Hideo
H
Sagawa
Yukiko
Y
Ohkusa
Toshifumi
T
Fujise
Kiyotaka
K
Gong
Jianlin
J
Tajiri
Hisao
H
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
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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