Murine respiratory tract dendritic cells: isolation, phenotyping and functional studies
Murine respiratory tract dendritic cells: isolation, phenotyping and functional studies
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DOI：
10.1016/j.jim.2004.01.019
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发表时间：
2004-04-01
影响因子：
2.2
通讯作者：
Dick, AD
中科院分区：
文献类型：
--
作者：
Calder, CJ;Liversidge, J;Dick, AD
呼吸道树突状细胞(RTDC)在鼻呼吸道内形成一个相邻的上皮下网络，连接天然免疫和获得性免疫，并参与鼻黏膜耐受的诱导。在RTDC的表型和功能方面，分离技术之间存在差异。因此，本研究的目的是改进以往的方法，以提供一种一致的分离方法，同时保持良好的细胞活力和丰富的细胞数量，以便于进一步的表型和功能的研究。经酶消化、Percoll密度梯度离心和GM-CSF过夜培养后分离的RTDC保持典型的未成熟树突状细胞表型，其特征是MHCIIlow CD40(Neg)CD86(Neg)CD80(Neg)CD11c(Low)细胞表面表达。分离的脾来源DC(SDC)符合体外培养第1天的表型；MHCIIlow CD40(Neg)CD86(Low)CD80(Neg)CD11c(Low)，但在体外可进一步成熟表型。RTDC和SDC在体外均能有效地处理抗原并将抗原递呈给T细胞。利用这种改良的RTDC分离方法，我们开发了一种一致的RTDC浓缩方法，它保持了RTDC的未成熟表型和功能抗原提呈能力。(C)2004爱思唯尔B.V.保留所有权利。
Respiratory tract dendritic cells (RTDC) form a contiguous subepithelial network within the nasorespiratory tract bridging innate and acquired immunity and have been implicated in nasal mucosal tolerance induction. Discrepancies exist between isolation techniques with respect to phenotype and function of RTDC. Therefore, the aim of this study was to modify previous methods to provide a consistent isolation method whilst maintaining good cell viability and enriched cell numbers so as to facilitate further phenotype and functional studies of murine RTDC. RTDCs isolated by enzyme digestion, Percoll density gradient centrifugation and overnight GM-CSF culture followed by MACS separation retain an archetypical immature dendritic cell phenotype, characterised by MHCIIlow CD40(neg) CD86(neg) CD80(neg) CD11c(low) cell surface expression. Splenic-derived DC (SDC) isolated conformed to a day 1 in vitro phenotype; MHCIIlow CD40(neg) CD86(low) CD80(neg) CD11c(low) but can further mature phenotypically in vitro. Both RTDC and SDC processed and presented antigen efficiently to T cells in vitro. Using such modified isolation procedures for RTDCs, we have developed a consistent method of RTDC enrichment, which maintains the immature phenotype and functional antigen presenting capability. (C) 2004 Elsevier B.V. All rights reserved.