Investigation of involvement of monocyte subpopulation in pathogenesis of sarcoidosis

CD 14 ^+ CD 16 ^+单核细胞是具有抗原呈递活性的单核细胞亚群，已知在炎性和感染性疾病如结核病、AIDS和川崎中数量增加。为了解单核细胞亚群在结节病发病中的作用，对CD 14 ^+ CD 16 ^+细胞的诱导机制、特性及功能进行了研究。使用单核细胞的磁分离系统和随后的流式细胞术方法，通过新鲜分离的单核细胞没有收获足够数量的细胞用于功能分析。因此，我们尝试用几种细胞因子组合从单核细胞中诱导CD 14 ^+ CD 16 ^+细胞。尽管先前报道的CD 14 ^+ CD 16 ^+细胞诱导因子MSCF/IL-4或TGFβ不能诱导大量的细胞，但MCSF/TGFβ或MSCF/IL 10强烈诱导CD 14 ^+ CD 16 ^+细胞。使用涂有聚四氟乙烯的盘子，而不是普通的盘子，也有利于诱导， ...更多信息 这是因为非贴壁细胞易于维持。另一方面，前列腺素E2显著抑制MSCF/IL-10的诱导。在前列腺素E_2受体中，EP_2与诱导有关。细胞因子诱导的CD 14 ^+ CD 16 ^+细胞表达CD 80而不表达CD 1a。趋化因子受体的表达为CXCR^-、CCR 4 ^<弱+>、CCR 5 ^<++>、CCR 6 ^<弱+>。在M-CSF/IL-10诱导的CD 14 ^+ CD 16 ^+细胞产生细胞因子方面，MDP和痤疮杆菌诱导了IL-1和IL-6的大量产生。为了了解Notch信号在诱导CD 14 ^+ CD 16 ^+细胞中的作用，我们使用了Notch配体包被的培养皿。在jagged-1或delta-1包被的培养皿中，这种诱导作用受到明显抑制，表明Notch信号与CD 14 ^+ CD 16 ^+细胞有关。在上皮样细胞诱导实验中，在Teflon包被的培养皿中长时间培养的单核细胞不显示上皮样细胞的特征。类上皮细胞也没有诱导维生素D3刺激或在纤连蛋白包被的培养皿。少

CD14^+CD16^+ monocytes have been implicated as a subpopulation of monocytes possessing an antigen presenting activity and known to be increased in number in inflammatory and infectious disorders such as tuberculosis, AIDS, and Kawasaki disease. To know the role of monocyte subpopulation in pathogenesis of sarcoidosis, the induction mechanism, characteristics and function of CD14^+CD16^+ cells were examined. Enough number of cells for function analysis was not harvested by freshly isolated mononuclear cells, using a magnet isolation system of monocytes and subsequent flowcytometry method. Therefore CD14^+CD16^+ cells were tried to be induced from monocytes by several kinds of cytokine combinations. Although MSCF/IL-4 or TGFβ, a previously reported induction factor of CD14^+CD16^+cells, did not induce a large number of the cells, MCSF/TGFβ or MSCF/IL10 strongly induced CD14^+CD16^+cells. To use teflon-coated dishes, instead of regular dishes, was also beneficial for the induction, probab … More ly because nonadherent cells were easily maintained. On the other hand, prostaglandin E2 significantly inhibited the induction by MSCF/IL-10. Among prostaglandin E_2 receptors, EP2 was associated with the induction. CD14^+CD16^+cells induced by the cytokines expressed CD80 but not CD1a. The expressions of chemokine receptors were CXCR^-, CCR4^<weak+>, CCR5^<++>, CCR6^<weak+>. In regard of the production of cytokines by M-CSF/IL-10-induced CD14^+CD16^+cells, MDP and p acnes induced high production of IL-1 and IL-6. To understand the involvement of Notch signals in the induction of CD14^+CD16^+cells, Notch ligand-coated dishes were used. The induction was significantly inhibited in jagged-1- or delta-1-coated dishes, suggesting that Notch signals was associated with CD14^+CD16^+ cells In regard of the experiment of epithelioid cell induction, monocytes cultured in teflon-coated dishes for long time did not show the features of epithelioid cells. Epithelioid cells were also not induced by vitamin D3 stimulation or in fibronectin-coated dishes. Less