The actin-bundling protein fascin is involved in the formation of dendritic processes in maturing epidermal Langerhans cells.
The actin-bundling protein fascin is involved in the formation of dendritic processes in maturing epidermal Langerhans cells.
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DOI：
10.4049/jimmunol.160.8.3776
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发表时间：
1998-04
影响因子：
4.4
通讯作者：
Ralf Ross;X. Ross;J. Schwing;T. Längin;Angelika B. Reske-Kunz
中科院分区：
文献类型：
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作者：
Ralf Ross;X. Ross;J. Schwing;T. Längin;Angelika B. Reske-Kunz
树突状细胞(DC)具有启动初始T细胞的独特潜能。表皮朗格汉斯细胞(LC)是存在于表皮中的树突状细胞(DC)，在体内或体外培养表皮细胞悬液时，可获得这种免疫刺激能力。为了分析成熟LC中的差异基因表达，我们构建了一个具有高度代表性的Lc文库，该文库包含18×10(6)个独立克隆。用新分离的朗格汉斯细胞(FLC)和CLC衍生的CLC特异性cDNA探针对该文库进行筛选。对鉴定出的cDNA进行测序，并通过数据库搜索进行分析。经鉴定，有两个片段为Fascin基因片段，表明Fascin基因在LC中存在差异表达。竞争性RT-PCR结果表明，在培养1、2、3天的CLC中高表达Fascin，而在FLC中未见表达。Western印迹和免疫荧光分析也表明Fasin在蛋白水平上具有CLC特异性表达。已知筋膜蛋白参与神经生长锥体胞质延伸中肌动蛋白细胞骨架的组织。其在成熟LC中的差异表达与LC中大量树突状突起的形成相一致。在培养过程中，LC与Fasin反义寡核苷酸孵育可抑制它们的形成。因此，我们得出结论，在成熟的朗格汉斯细胞的树突状突起的形成过程中，发束素是必需的，并因此可能影响T细胞与LC的相互作用。
Dendritic cells (DC) are characterized by their unique potential to prime naive T cells. Epidermal Langerhans cells (LC), the DC resident in the epidermis, gain this immunostimulatory capacity following Ag contact in vivo or during in vitro culture of epidermal cell suspensions. To analyze differential gene expression in maturing LC, we constructed a highly representative cDNA library of cultivated LC (cLC) in lambda ZAP II containing 18 x 10(6) independent clones. This library was screened with freshly isolated Langerhans cell (fLC)- and cLC-derived probes for cLC-specific cDNAs. The cDNAs identified were sequenced and analyzed by database searches. Two cDNA fragments were identified as fragments of fascin, indicating that fascin is differentially expressed in LC. By competitive RT-PCR, we confirmed that fascin is highly expressed in cLC cultivated for 1, 2, and 3 days, while no signals were obtained with fLC. Western blot and immunofluorescence analysis revealed cLC-specific expression of fascin on the protein level as well. Fascin is known to be involved in the organization of the actin cytoskeleton in cytoplasmatic extensions of nerve growth cones. Its differential expression in maturing LC coincides with the formation of numerous dendritic projections in LC. Their formation was inhibited by incubation of LC with fascin antisense oligonucleotides during cultivation. Therefore, we conclude that fascin is necessary for the formation of the dendritic processes of maturing Langerhans cells and may thus influence T cell-LC interaction.