T helper cell type 2 cytokine-mediated comitogenic responses and CCR3 expression during differentiation of human mast cells in vitro.
T helper cell type 2 cytokine-mediated comitogenic responses and CCR3 expression during differentiation of human mast cells in vitro.
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DOI：
10.1084/jem.190.2.267
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发表时间：
1999-07-19
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肥大细胞（MC）原位产生于循环干细胞因子（SCF）依赖性定向祖细胞（PrMC），并在过敏性粘膜炎症部位积聚。我们假设人类（h）PrMC及其成熟对应物可能与嗜酸性粒细胞、嗜碱性粒细胞和辅助性T细胞2型（Th 2）淋巴细胞共享趋化因子和细胞因子受体利用的重叠模式，用于其归巢和过敏相关增生。我们已经确定了来自脐带血的定向hPrMCs和完全成熟的hMCs对与过敏性炎症密切相关的趋化因子和细胞因子激动剂的功能性反应及其相应受体的成熟相关表达的特征。在存在重组干细胞因子（SCF）、白细胞介素（IL）-6和IL-10的情况下培养4周后，基于其c-kit和CD 13的均匀表面表达、Fc∈RIα的低水平表达、CD 14和CD 16的不表达以及MC糜酶的免疫反应性> 80%，约一半为类胰蛋白酶免疫反应阳性，甲苯胺蓝异染。到第9周，细胞已成熟为hMC，通过较高水平的c-kit、持续表达CD 13和低水平的Fc∈RIα、均匀的甲苯胺蓝异染性以及均匀的类胰蛋白酶和糜蛋白酶免疫反应性来鉴定。4周龄的hPrMCs表达四种趋化因子受体（CXCR 2、CCR 3、CXCR 4和CCR 5）。每个受体介导的短暂快速钙流响应其各自的配体。重组人eotaxin和基质细胞衍生因子1α均能诱导hPrMCs的趋化性。只有CCR 3保留在这些趋化因子受体中的成熟9周龄hMC上，hMC对嗜酸性粒细胞趋化因子有持续的钙流反应，但没有趋化性。Th 2细胞因子IL-3、IL-5、IL-6、IL-9和粒细胞/巨噬细胞集落刺激因子均增强hPrMC和hMC的SCF依赖性增殖。相反，典型的Th 1细胞因子干扰素γ抑制SCF驱动的hPrMCs和hMCs的增殖。因此，在其整个体外发育过程中，hMCs服从SCF依赖性、烟碱驱动的促有丝分裂反应，反映了过敏和哮喘的Th 2型极化特征。此外，定型hPrMCs具有来自所报道的造血细胞的趋化因子受体表达的独特谱，包括CCR 3，其与过敏性炎症的其他细胞（嗜酸性粒细胞、嗜碱性粒细胞和Th 2淋巴细胞）共享。
Mast cells (MCs) arise in situ from circulating stem cell factor (SCF)-dependent committed progenitors (PrMCs) and accumulate at sites of allergic mucosal inflammation. We hypothesized that human (h)PrMCs and their mature counterparts might share overlapping patterns of chemokine and cytokine receptor utilization with eosinophils, basophils, and T helper type 2 (Th2) lymphocytes for their homing and allergy-associated hyperplasia. We have characterized committed hPrMCs and fully mature hMCs derived in vitro from cord blood for their functional responses to chemokine and cytokine agonists germane to allergic inflammation and for their maturation-related expression of the corresponding receptors. After 4 wk of culture in the presence of recombinant stem cell factor (SCF), interleukin (IL)-6, and IL-10, the cells were characterized as hPrMCs based upon their uniform surface expression of c-kit and CD13, low-level expression of Fc∈RIα, absence of CD14 and CD16 expression, and immunoreactivity for MC chymase in >80%, and about half were immunoreactive for tryptase and metachromatic with toluidine blue. By week 9, the cells had matured into hMCs, identified by higher levels of c-kit, continued expression of CD13 and low-level Fc∈RIα, uniform toluidine blue metachromasia, and uniform immunoreactivity for both tryptase and chymase. The 4-wk-old hPrMCs expressed four chemokine receptors (CXCR2, CCR3, CXCR4, and CCR5). Each receptor mediated transient rapid calcium fluxes in response to its respective ligand. Both recombinant human eotaxin and stromal cell–derived factor 1α elicited chemotaxis of hPrMCs. Only CCR3 was retained on the mature 9-wk-old hMCs from among these chemokine receptors, and hMCs responded to eotaxin with a sustained calcium flux but without chemotaxis. The Th2 cytokines IL-3, IL-5, IL-6, IL-9, and granulocyte/macrophage colony-stimulating factor each augmented the SCF-dependent proliferation of hPrMCs and hMCs. In contrast, the prototypical Th1 cytokine, interferon γ, suppressed SCF-driven proliferation of both hPrMCs and hMCs. Thus, throughout their development in vitro, hMCs obey SCF-dependent, cytokine-driven mitogenic responses that reflect a Th2-type polarization characteristic of allergy and asthma. Furthermore, committed hPrMCs have a unique profile of chemokine receptor expression from among reported hematopoietic cells, including CCR3, which is shared with the other cells central to allergic inflammation (eosinophils, basophils, and Th2 lymphocytes).