Identification and clinical application for the role of NKT cells in patients with chronic hepatitis and hepatocellualr carcinoma infected hepatitis virus

NKT细胞在肝炎病毒感染的慢性肝炎及肝癌患者中的作用鉴定及临床应用

基本信息

批准号：
15390236
负责人：
KAKUMU Shinichi
金额：
$ 9.09万
依托单位：
Aichi Medical University School of Medicine
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
2003
资助国家：
日本
起止时间：
2003 至 2005
项目状态：
已结题

项目摘要

Natural killer T (NKT) cells share features of both classical T cells and NK cells. NKT cells are heterogenous populations, and recognize glycolipids associated with CD1d molecule. We investigated Th1/Th2 cytokine production as well as frequency and phenotype of circulating NKT cells in patients with chronic hepatitis (CH) and hepatocellular carcinoma (HCC) infected with hepatitis C virus (HCV). Peripheral blood mononuclear cells (PBMC) were obtained befor and 2 weeks later IFN/ribavirin and radiofrequency ablation therapy for CH and HCC, respectively. PBMC were cultured with α galactosylceramide (α GalCer) and IL-2. Frequency and cytokine production of NKT cells were analyzed with flow cytometry. IFN-γ production of Vα 24+CD3+ T cells did not differ among groups, but became greater after treatment in contrast to lowered IL-4 production.The function of dendritic cells (DCs) appears to be decreased in patients with chronic hepatitis C. However, we have no strategy to modulate its functi … More on. Thus we wanted to induce maturated DC by activating V α 24+ NKT cells. We cultured peripheral blood CD14+ cells in the presence of GMCSF and IL-4. Simultaneously CD3+ T cells were cultured with α GalCer + IL-2. After 5-day culture, they are mixed and cultured with GMCSF, α GalCer and IL-2 for further 2 days. This method induced matured, functional DC.Cytotoxic T lymphocyes (CTLs) are thought to be major effectors for clearing viruses in acute infections including hepatitis B virus (HBV). Persistent HBV infection is characterized by a lack of or a weak CTL response to HBV, which is thought to reflect tolerance to HBV antigens. We found that α GalCer, a ligand for V α 14+ NKT cells in mice, strongly enhanced the induction and proliferation of HBV-specific CTLs by HBsAg. In HbsAg transgenic mice, which are thought to be tolerant to HBV-encoded antigens, administration of HbsAg or α GalCer alone failed to HbsAg-specific CTLs, but they were induced by administration of both compounds. A blocking experiment using antibodies to cytokines and CD4O ligand showed that IL-2 mediates the enhancement of CTL induction caused by α GalCer through NKT activation. Less

天然杀伤T（NKT）细胞具有经典T细胞和NK细胞的特征。 NKT细胞是异质种群，并且识别与CD1D分子相关的糖脂。我们研究了慢性肝炎（CH）和肝细胞癌（HCC）感染丙型肝炎病毒（HCV）的慢性肝炎（CH）和肝细胞癌（HCV）患者的TH1/TH2细胞因子的产生以及循环NKT细胞的频率和表型。分别为CH和HCC的IFN/Ribavirin和Radiofrequency消融疗法获得外周血单核细胞（PBMC）。将PBMC用α甲乳糖基酰胺（αGalcer）和IL-2培养。通过流式细胞仪分析NKT细胞的频率和细胞因子的产生。 Vα24+ CD3+ T细胞的IFN-γ产生在组之间没有差异，但与IL-4的产生相比，治疗后变得更大。树突状细胞（DC）的功能（DC）在慢性丙型肝炎患者中似乎有所改善。但是，我们没有调节其功能的策略。我们想通过激活Vα24+NKT细胞来诱导成熟的DC。在GMCSF和IL-4存在下，我们培养了外周血CD14+细胞。同时用αGalcer + IL-2培养CD3 + T细胞。 5天培养后，将它们与GMCSF，αGalcer和IL-2一起培养2天。这种方法诱导的成熟，功能性DC。胞毒性T淋巴细胞（CTL）被认为是清除包括丙型肝炎病毒（HBV）的急性感染病毒的主要作用。持续性HBV感染的特征是缺乏对HBV的CTL反应，这被认为反映了对HBV抗原的耐受性。我们发现，小鼠中Vα14+NKT细胞的配体αGalcer强烈增强了HBSAG对HBV特异性CTL的诱导和增殖。在HBSAG转基因小鼠中，被认为对HBV编码的抗原具有耐受性，单独使用HBSAG或αgalcer的抗原无法使用HBSAG特异性CTL，但它们是通过两种化合物的给药而诱导的。使用对细胞因子和CD4O配体抗体的阻断实验表明，IL-2介导了通过NKT激活引起的αgalcer引起的CTL诱导的增强。较少的