Morelli A, Larregina A, Chuluyán I, Kolkowski E, Fainboim L
Laboratory of Immunogenetics, Argentina, USA.
Immunology. 1996 Sep;89(1):126-34. doi: 10.1046/j.1365-2567.1996.d01-701.x.
Although it is known that dendritic cells (DC) migrate in response to inflammatory stimuli. There is little information about the expression of receptors for chemotactic factors on DC. The present study has demonstrated by double immunostaining and flow cytometry of Langerhan's cell (LC)-enriched epidermal cell suspensions that a small subpopulation (5-6%) of epidermal resident DC (rLC) expresses receptors for C5a (C5aR). Epidermal rLC positive for C5aR show a round-shape morphology, were located next to the basement membrane and express HLA-DR molecules higher than C5aR negative rLC. These observations suggest that rLC would express C5aR as part of their process of maturation during tissue trafficking. To investigate whether epidermal LC up-regulate C5aR along their differentiation pathway. LC were differentiated in vitro after culture in epidermal cell suspensions supplemented with granulocyte macrophage colony-stimulating factor (GM-CSF). As a result, in vitro differentiated LC increased the expression of C5aR up to 69% of the DC population. In accordance with this observation, interdigitating DC of secondary lymphoid organs (lymph node and tonsil) also expressed (5aR. Migratory CD1a positive DC that spontaneously migrated out of dermal or split-skin organ explants were also positive for C5aR and were used for chemotaxis and chemokinesis assays in response to human recombinant C5a (rC5a). Optimum migration to rC5a was observed at 10(-8)M with a sigmoidal dose response curve. Checkboard analysis demonstrated that locomotion in response to rC5a was chemotaxis and not chemokinesis.
尽管已知树突状细胞(DC)会响应炎症刺激而迁移,但关于DC上趋化因子受体的表达情况却知之甚少。本研究通过对富含朗格汉斯细胞(LC)的表皮细胞悬液进行双重免疫染色和流式细胞术证明,一小部分(5 - 6%)表皮驻留DC(rLC)表达C5a受体(C5aR)。C5aR阳性的表皮rLC呈圆形形态,位于基底膜附近,且表达的HLA - DR分子高于C5aR阴性的rLC。这些观察结果表明,rLC在组织迁移过程中作为其成熟过程的一部分会表达C5aR。为了研究表皮LC在其分化途径中是否会上调C5aR,在补充粒细胞巨噬细胞集落刺激因子(GM - CSF)的表皮细胞悬液中培养后,体外诱导LC分化。结果,体外分化的LC使C5aR的表达增加,高达DC群体的69%。与此观察结果一致,次级淋巴器官(淋巴结和扁桃体)的交错突DC也表达C5aR。自发从真皮或劈开皮肤器官外植体迁移出的迁移性CD1a阳性DC对C5aR也呈阳性,并用于针对重组人C5a(rC5a)的趋化性和化学动力学分析。在10^(-8)M时观察到对rC5a的最佳迁移,呈S形剂量反应曲线。棋盘分析表明,对rC5a的运动是趋化性而非化学动力学。