Granulocyte/macrophage colony-stimulating factor is essential for the viability and function of cultured murine epidermal Langerhans cells.
Granulocyte/macrophage colony-stimulating factor is essential for the viability and function of cultured murine epidermal Langerhans cells.
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DOI：
10.1084/jem.166.5.1484
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发表时间：
1987-11-01
期刊：
影响因子：
--
通讯作者：
Steinman RM
中科院分区：
文献类型：
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作者：
Witmer-Pack MD;Olivier W;Valinsky J;Schuler G;Steinman RM
建立了一种从小鼠表皮中富集朗格汉斯细胞（LC）的方法。在标准培养基中，富集的群体在3-d间隔内逐渐失去活力。当培养物补充有角质形成细胞条件培养基时，LC活力得到改善，并且细胞的树突状突起的大小和数量增加。通过刺激混合淋巴细胞反应（MLR）的活性监测，辅助功能至少增加10- 20倍。刺激的巨噬细胞和T细胞的条件培养基也支持培养的LC的活力和成熟。一组纯化的细胞因子已经过测试，只有粒细胞/巨噬细胞集落刺激因子（GM-CSF）替代散装条件培养基。重组分子在5 pM时表现出半最大活性。没有活性的是：IL-1-4; IFN-α/β/γ;恶病质/TNF; M-和G-CSF。兔抗GM-CSF特异性中和角质形成细胞条件培养基产生活性LC的能力。然而，在MLR本身期间，LC功能不需要GM-CSF。我们的结论是免疫活性LC在文化的发展是由GM-CSF介导的。这些树突状细胞不响应谱系特异性G-和M-CSF的观察表明，LC代表一个独特的髓样分化途径。因为GM-CSF可以由非免疫细胞产生，并且可以介导活性树突状细胞的产生，所以这种细胞因子提供了用于增强细胞介导的免疫的致敏阶段的T非依赖性机制。
A panning method has been developed to enrich Langerhans cells (LC) from murine epidermis. In standard culture media, the enriched populations progressively lose viability over a 3-d interval. When the cultures are supplemented with keratinocyte-conditioned medium, LC viability is improved and the cells increase in size and number of dendritic processes. Accessory function, as monitored by stimulating activity in the mixed lymphocyte reaction (MLR), increases at least 10- 20-fold. The conditioned media of stimulated macrophages and T cells also support the viability and maturation of cultured LC. A panel of purified cytokines has been tested, and only granulocyte/macrophage colony-stimulating factor (GM-CSF) substitutes for bulk-conditioned medium. The recombinant molecule exhibits half-maximal activity at 5 pM. Without activity are: IL-1-4; IFN-alpha/beta/gamma; cachectin/TNF; M- and G-CSF. A rabbit anti-GM-CSF specifically neutralizes the capacity of keratinocyte-conditioned medium to generate active LC. However, GM-CSF is not required for LC function during the MLR itself. We conclude that the development of immunologically active LC in culture is mediated by GM-CSF. The observation that these dendritic cells do not respond to lineage-specific G- and M-CSFs suggests that LC represent a distinct myeloid differentiation pathway. Because GM-CSF can be made by nonimmune cells and can mediate the production of active dendritic cells, this cytokine provides a T-independent mechanism for enhancing the sensitization phase of cell-mediated immunity.