In vitro generation of cytotoxic and regulatory T cells by fusions of human dendritic cells and hepatocellular carcinoma cells
In vitro generation of cytotoxic and regulatory T cells by fusions of human dendritic cells and hepatocellular carcinoma cells
复制标题
DOI：
10.1186/1479-5876-6-51
复制
发表时间：
2008-09-15
影响因子：
7.4
通讯作者：
Tajiri, Hisao
中科院分区：
文献类型：
--
作者：
Koido, Shigeo;Homma, Sadamu;Tajiri, Hisao
背景：人肝细胞癌（HCC）细胞表达WT1和/或癌胚抗原（CEA）作为诱导抗肿瘤免疫的潜在靶点。在这项研究中，我们检测了树突状细胞（DCs）和HCC细胞融合后产生的细胞毒性T淋巴细胞（CTL）和调节性T细胞（Treg）。方法：将健康供者或HCC患者的HCC细胞与DCs融合，研究HCC细胞培养（HCCsp）的上清液是否影响DCs/HCC融合细胞（fc）的功能以及CTL和Treg的产生。结果：fc共表达HCC来源的WT1和CEA抗原以及dcs来源的MHC II类和共刺激分子。此外，FCs有效激活CD4（+）和CD8(+) T细胞，这些T细胞能够产生ifn - γ，并通过MHC i类限制机制诱导自身肿瘤或半同种异体靶点的细胞溶解。然而，HCCsp通过下调MHC I类和II类、CD80、CD86和CD83分子来诱导dc的功能损伤。此外，暴露于hccsp的dc在toll样受体4激动剂青霉素灭活的化脓性链球菌刺激下不能完全成熟。有趣的是，在HCCsp存在下产生的未成熟dc和异体HCC细胞的融合促进了CD4(+) CD25（高）Foxp3(+) Treg的产生，并抑制了HCCsp存在下CTL的诱导。重要的是，在晚期HCC患者接种自体fc后，观察到dc上MHC II类、CD80和CD83的上调。此外，FCs诱导的WTI和cea特异性CTL能够产生高水平的ifn - γ。结论：本研究首次证实了dc和HCC细胞融合诱导抗原特异性CTL和Treg的产生。局部肿瘤相关因素可能通过抑制dc成熟促进Treg的生成；然而，融合细胞接种可在体外恢复dc功能并诱导抗原特异性CTL反应。本研究可能为FCs产生CTL和Treg的机制提供新的思路。
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(+) 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 WTI- and CEA-specific CTL that were able to produce high levels of IFN-gamma.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.