Laminin and fibronectin treatment leads to generation of dendritic cells with superior endocytic capacity.
Laminin and fibronectin treatment leads to generation of dendritic cells with superior endocytic capacity.
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DOI：
10.1371/journal.pone.0010123
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发表时间：
2010-04-19
期刊：
影响因子：
3.7
通讯作者：
Ghaemmaghami AM
中科院分区：
文献类型：
--
作者：
García-Nieto S;Johal RK;Shakesheff KM;Emara M;Royer PJ;Chau DY;Shakib F;Ghaemmaghami AM
树突状细胞（DC）在微生物暴露部位的微环境采样及其随后与淋巴结副皮质区T细胞的相互作用是启动免疫应答的关键事件。我们对人体中此类事件的大多数了解是基于在细胞外基质（ECM）蛋白不存在的情况下进行的体外研究。不同组织（包括淋巴器官）的基底膜和间质空间中的ECM在控制特定细胞功能（如迁移、细胞内信号传导和分化）中起重要作用。因此，本研究的目的是调查两种丰富的ECM成分，纤连蛋白和层粘连蛋白，对DC的表型和功能特性的影响，以及这可能如何影响DC诱导的T细胞分化。用层粘连蛋白和纤连蛋白处理人单核细胞来源的DC长达48小时，并使用扫描电子显微镜、流式细胞术和真实的时间PCR分析其形态和表型。流式细胞术检测DC的内吞能力。建立DC与T细胞共培养体系，分别采用H3-胸苷掺入法和ELISA法检测T细胞增殖和细胞因子水平。最后，我们使用不同的细胞追踪器和流式细胞术评估DC-T细胞缀合物的形成。我们的数据表明，在ECM的存在下，DC保持“更不成熟”的表型，并表达更高水平的关键内吞受体，因此成为显著更好的内吞细胞，但仍然能够完全成熟，以响应刺激，如其诱导抗原特异性T细胞分化的上级能力所证明的。这些研究强调了在研究DC生物学和DC-T细胞相互作用的体外研究中包括ECM组分的重要性。在抗原特异性DC诱导的T细胞增殖的背景下，包含ECM蛋白可以导致开发更灵敏的测定。
Sampling the microenvironment at sites of microbial exposure by dendritic cells (DC) and their subsequent interaction with T cells in the paracortical area of lymph nodes are key events for initiating immune responses. Most of our knowledge of such events in human is based on in vitro studies performed in the absence of extracellular matrix (ECM) proteins. ECM in basement membranes and interstitial spaces of different tissues, including lymphoid organs, plays an important role in controlling specific cellular functions such as migration, intracellular signalling and differentiation. The aim of this study was, therefore, to investigate the impact of two abundant ECM components, fibronectin and laminin, on the phenotypical and functional properties of DC and how that might influence DC induced T-cell differentiation. Human monocyte derived DC were treated with laminin and fibronectin for up to 48 hours and their morphology and phenotype was analyzed using scanning electron microscopy, flow cytometry and real time PCR. The endocytic ability of DC was determined using flow cytometry. Furthermore, co-culture of DC and T cells were established and T cell proliferation and cytokine profile was measured using H3-thymidine incorporation and ELISA respectively. Finally, we assessed formation of DC-T cell conjugates using different cell trackers and flow cytometry. Our data show that in the presence of ECM, DC maintain a ‘more immature’ phenotype and express higher levels of key endocytic receptors, and as a result become significantly better endocytic cells, but still fully able to mature in response to stimulation as evidenced by their superior ability to induce antigen-specific T cell differentiation. These studies underline the importance of including ECM components in in vitro studies investigating DC biology and DC-T cell interaction. Within the context of antigen specific DC induced T cell proliferation, inclusion of ECM proteins could lead to development of more sensitive assays.
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影响因子：
4.8
作者：
Frison, N;Taylor, ME;Roche, AC
通讯作者：
Roche, AC
影响因子：
5.4
作者：
Ardouin, L;Bracke, M;Tybulewicz, VLJ
通讯作者：
Tybulewicz, VLJ
影响因子：
5.5
作者：
Gorfu, Gezahegn;Virtanen, Ismo;Patarroyo, Manuel
通讯作者：
Patarroyo, Manuel
影响因子：
--
作者：
HAYAKAWA, M;KOBAYASHI, M;HOSHINO, T
通讯作者：
HOSHINO, T
影响因子：
4.4
作者：
Andersen, Claire A. Swetman;Handley, Matthew;Chain, Benjamin M.
通讯作者：
Chain, Benjamin M.