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

自然杀伤T（NKT）细胞具有经典T细胞和NK细胞的特征。NKT细胞是一种异质性细胞，识别与CD 1d分子相关的糖脂。我们研究了Th 1/Th 2细胞因子的产生以及循环NKT细胞的频率和表型在慢性肝炎（CH）和肝细胞癌（HCC）感染丙型肝炎病毒（HCV）的患者。分别在干扰素/利巴韦林和射频消融治疗CH和HCC前和治疗2周后采集外周血单个核细胞（PBMC）。用α半乳糖神经酰胺（α GalCer）和IL-2培养PBMC。用流式细胞术分析NKT细胞的频率和细胞因子产生。慢性丙型肝炎患者外周血中Vα 24+ CD 3 + T细胞IFN-γ的分泌无明显差异，但治疗后增加，而IL-4的分泌减少。然而，我们没有调整其功能的策略， ...更多信息 因此，我们希望通过激活V α 24+ NKT细胞来诱导成熟DC。我们培养的外周血CD 14+细胞的存在下，GMCSF和IL-4。同时用α GalCer + IL-2培养CD 3 + T细胞。培养5天后，将其与GMCSF、α GalCer和IL-2混合并再培养2天。细胞毒性T淋巴细胞（CTL）被认为是清除包括B型肝炎病毒（HBV）在内的急性感染中的病毒的主要效应物。持续性HBV感染的特征在于缺乏或弱的针对HBV的CTL应答，这被认为反映了对HBV抗原的耐受性。我们发现，α GalCer（小鼠V α 14+ NKT细胞的配体）强烈增强HBsAg对HBV特异性CTL的诱导和增殖。在被认为对HBV编码抗原耐受的HBsAg转基因小鼠中，单独给予HBsAg或α GalCer不能诱导HBsAg特异性CTL，但给予两种化合物可诱导CTL。使用针对细胞因子和CD 4 O配体的抗体的阻断实验表明，IL-2通过NKT活化介导α GalCer引起的CTL诱导的增强。少

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