Detection and activation of human Vα24+ natural killer T cells using α-galactosyl ceramide-pulsed dendritic cells
Detection and activation of human Vα24+ natural killer T cells using α-galactosyl ceramide-pulsed dendritic cells
复制标题
DOI：
10.1016/s0022-1759(02)00497-0
复制
发表时间：
2003-01-15
影响因子：
2.2
通讯作者：
Dhodapkar, MV
中科院分区：
文献类型：
--
作者：
Fujii, S;Shimizu, K;Dhodapkar, MV
人CD 1d限制性自然杀伤T（NKT）细胞被认为在几种临床环境中调节免疫应答，可被抗原呈递细胞（APC）上CD 1d分子呈递的β-半乳糖神经酰胺（（x-GalCer））激活。量化新鲜血液中NKT功能的简单方法将极大地有利于针对人类NKT细胞的研究。在这里，我们表明，新鲜分离的人NKT细胞可以很容易地定量的酶联免疫斑点（ELISPOT）测定，并有一个Th 1型（分泌干扰素-γ，但不是IL-4），刺激后使用α-GalCer加载的APC。使用该测定，我们还评估了新鲜血液中人NKT细胞活化的APC要求。单核细胞衍生的树突状细胞（DC）比单核细胞/巨噬细胞更有效地检测和激活新鲜血液中的NKT细胞，其中成熟的α-GalCer脉冲的DC是最佳的。DC也是用于扩增培养物中的NKT细胞和产生NKT细胞系的有效APC。用DC扩增的NKT细胞是功能性的，分泌IFN-γ和IL-4，并杀死NKT敏感的靶标。使用加载有10-100 ng/ml α-GalCer的成熟DC观察到这些细胞系的最佳活化。用几种不同的刺激物成熟的DC是有效的。这些数据有助于建立用α-GalCer负载DC用于NKT细胞的免疫治疗靶向的条件，并提供了一种新的简单测定来监测人类中的NKT功能。(C)2002 Elsevier Science B. V.保留所有权利。
Human CD1d-restricted natural killer T (NKT) cells, which are postulated to regulate the immune response in several clinical settings, can be activated by (x-galactosylceramide ((x-GalCer) presented by CD1d molecules on antigen presenting cells (APCs). Simple methods to quantify NKT function in fresh blood will greatly benefit studies targeting NKT cells in humans. Here we show that freshly isolated human NKT cells can be readily quantified by an enzyme-linked immunospot (ELISPOT) assay and have a Th1 profile (secreting interferon-gamma, but not IL-4), after stimulation using alpha-GalCer loaded APCs. Using this assay, we also evaluated APC requirements for human NKT cell activation in fresh blood. Monocyte-derived dendritic cells (DCs) are more effective than monocytes/macrophages for detecting and activating NKT cells in fresh blood, with mature alpha-GalCer pulsed DCs being optimal. DCs are also efficient APCs for expanding NKT cells in culture and generating NKT cell lines. NKT cells expanded with DCs were functional, secreting both IFN-gamma and IL-4, and killing NKT-sensitive targets. Optimal activation of these lines was seen using mature DCs loaded with 10-100 ng/ml of alpha-GalCer. DCs matured with several different stimuli were effective. These data help to establish the conditions for loading DCs with alpha-GalCer for immune therapeutic targeting of NKT cells, and provide a new simple assay to monitor NKT function in humans. (C) 2002 Elsevier Science B.V. All rights reserved.