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

自然杀伤 T (NKT) 细胞具有经典 T 细胞的特征，NKT 细胞是异质群体，并且识别与 CD1d 分子相关的糖脂。我们研究了 Th1/Th2 细胞因子的产生以及循环 NKT 细胞的频率和表型。感染丙型肝炎病毒（HCV）的慢性肝炎（CH）和肝细胞癌（HCC）患者在治疗前和治疗后2周获取外周血单核细胞（PBMC）。随后分别用α半乳糖神经酰胺(α GalCer)和IL-2对CH和HCC进行干扰素/利巴韦林和射频消融治疗，并通过流式细胞术分析NKT细胞的细胞因子产生。 +CD3+ T 细胞在各组之间没有差异，但在治疗后数量增加，而 IL-4 产量降低。树突状细胞 (DC) 的功能似乎慢性丙型肝炎患者中的 CD14+ 细胞会减少。然而，我们没有调节其功能的策略，因此我们希望通过激活 V α 24+ NKT 细胞来诱导成熟的 DC，并在 GMCSF 存在的情况下培养外周血 CD14+ 细胞。将CD3+T细胞与αGalCer+IL-2同时培养。培养5天后，将它们与GMCSF、αGalCer和IL-4混合培养。 IL-2 再持续 2 天，该方法诱导成熟的功能性 DC。细胞毒性 T 淋巴细胞 (CTL) 被认为是清除急性感染（包括乙型肝炎病毒 (HBV)）中病毒的主要效应物。对 HBV 的 CTL 反应较弱或较弱，这被认为反映了对强 HBV 抗原的耐受性。我们发现，小鼠中 V α 14+ NKT 细胞的配体 α GalCer 强烈增强了诱导作用。在被认为对 HBV 编码抗原具有耐受性的 HbsAg 转基因小鼠中，单独施用 HbsAg 或 α GalCer 无法促进 HbsAg 特异性 CTL 的增殖，但同时施用两种化合物都可以诱导它们增殖。使用细胞因子抗体和 CD4O 配体进行的封闭实验表明，IL-2 通过 NKT 激活介导 α GalCer 引起的 CTL 诱导增强。