Research

In vitro generation of cytotoxic and regulatory T cells by fusions of human dendritic cells and hepatocellular carcinoma cells

Shigeo Koido^1,2 , Sadamu Homma³ , Eiichi Hara⁵ , Makoto Mitsunaga¹ , Yoshihisa Namiki² , Akitaka Takahara^1,3 , Eijiro Nagasaki³ , Hideo Komita¹ , Yukiko Sagawa⁴ , Toshifumi Ohkusa^1,2 , Kiyotaka Fujise^1,2 , Jianlin Gong⁶ and Hisao Tajiri¹

¹  Division of Gastroenterology and Hepatology, Department of Internal Medicine, The Jikei University School of Medicine, Tokyo, Japan

²  Institute of Clinical Medicine and Research, The Jikei University School of Medicine, Tokyo, Japan

³  Department of Oncology, Institute of DNA Medicine, The Jikei University School of Medicine, Tokyo, Japan

⁴  Clinical Data Bank, Institute of DNA Medicine, The Jikei University School of Medicine, Tokyo, Japan

⁵  Research Institute for Clinical Oncology, Saitama Cancer Center, Saitama, Japan

⁶  Department of Medicine, Boston University School of Medicine, Boston, MA, USA

author email corresponding author email

Journal of Translational Medicine 2008, 6:51doi:10.1186/1479-5876-6-51

Published:	15 September 2008

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-γ 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⁺ CD25^high 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-γ.

Conclusion

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.