Isolation of myeloid and plasmacytoid dendritic cells from human bronchoalveolar lavage fluid
Isolation of myeloid and plasmacytoid dendritic cells from human bronchoalveolar lavage fluid
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从人支气管肺泡灌洗液中分离骨髓和浆细胞样树突状细胞
DOI：
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发表时间：
2006
影响因子：
4
通讯作者：
N. Siafakas
中科院分区：
文献类型：
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作者：
M. Tsoumakidou;N. Tzanakis;H. Papadaki;Helen Koutala;N. Siafakas
支气管肺泡灌洗液（BALF）树突状细胞（DC）的研究一直受到DC稀缺和缺乏DC特异性表面标志物的阻碍。最近有四种表面抗原被描述为DC不同亚群的特异性标志物，并已用于从肺切除标本中分离和表征新鲜的非培养DC：BDCA-1（CD 1c）和BDCA-3分别用于髓样DC 1型和2型，BDCA-2和BDCA-4用于浆细胞样DC。本研究的目的是发展一种新的方法，用于分离支气管肺泡灌洗液DC，使用BDCA+细胞的免疫磁性分离。在Ficoll-Paque密度梯度离心后，从BALF中获得单核细胞。对单核细胞、T细胞和B细胞进行磁性标记并去除。将未标记的细胞部分与BDCA-1、BDCA-3和BDCA-4珠一起孵育，并保留总BDCA+ DC。通过将分离的DC与免疫磁性分选的初始T细胞共培养来评价分离的DC诱导T细胞应答的能力。上述程序产生了在诱导T细胞应答中显示出强能力的活DC群体。通过免疫磁性分离可以容易地获得功能完整的人BALF髓样DC 1型和2型以及浆细胞样DC。考虑到支气管肺泡灌洗是一个微创的程序，这些细胞是最佳的候选人，以阐明肺DC的属性和能力。
Studies of bronchoalveolar lavage fluid (BALF) dendritic cells (DC) have been hampered by the scarcity of DC and the lack of DC‐specific surface markers. Four surface Ag have been recently described as specific markers for distinct subsets of DC and have been used for the isolation and characterization of fresh noncultured DC from lung resection specimens: BDCA‐1 (CD1c) and BDCA‐3 for myeloid DC type 1 and type 2, respectively, and BDCA‐2 and BDCA‐4 for plasmacytoid DC. The aim of this study was to develop a new method for the isolation of BALF DC, using immunomagnetic separation of BDCA+ cells. Mononuclear cells were obtained from BALF after Ficoll‐Paque density gradient centrifugation. Monocytes, T cells and B cells were magnetically labelled and depleted. The unlabelled cell fraction was incubated with BDCA‐1, BDCA‐3 and BDCA‐4 beads and the total BDCA+ DC were retained. The ability of isolated DC to induce T‐cell responses was evaluated by coculturing the isolated DC with immunomagnetically sorted naive T cells. The above procedure resulted in a population of viable DC that showed a strong capacity in induce T‐cell responses. Functionally intact human BALF myeloid DC types 1 and 2 as well as plasmacytoid DC can be easily obtained by immunomagnetic isolation. Considering that bronchoalveolar lavage is a minimally invasive procedure, these cells are optimal candidates with which to elucidate the properties and capabilities of pulmonary DC.
DOI：
10.2741/1185
发表时间：
2003-09-01
期刊：
FRONTIERS IN BIOSCIENCE
影响因子：
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作者：
Donnenberg, VS;Donnenberg, AD
通讯作者：
Donnenberg, AD