Parker Jeremy C, Thavagnanam Surendran, Skibinski Grzegorz, McBrien Michael, Heaney Liam G, Shields Michael D
Centre for Infection and Immunity, Health Sciences Building, Queen's University Belfast, 97 Lisburn Road, Belfast BT9 7BL, Northern Ireland, United Kingdom.
Department of Anaesthetics, Belfast Health and Social Care Trust, Royal Victoria Hospital, Belfast BT12 6BE, Northern Ireland, United Kingdom.
Results Immunol. 2012 May 15;2:104-11. doi: 10.1016/j.rinim.2012.05.001. eCollection 2012.
IL-31 is a novel cytokine that has been implicated in allergic diseases such as atopic dermatitis and more recently asthma. While IL-31 has been well studied in skin conditions such as atopic dermatitis, little is known about the role IL-31 plays in asthma and specifically the differentiation process of the bronchial epithelium, which is central to the pathogenesis of allergic asthma.
We examined the effects of IL-13 (20 ng/ml), IL-31 (20 ng/ml) and an IL-13/IL-31 combination stimulation (20 ng/ml each) on the in vitro mucociliary differentiation of paediatric bronchial epithelial cells (PBECs) from healthy patients (n=6). IL-31 receptor (IL-31-RA) expression, markers of differentiation (goblet and ciliated cells), transepithelial electrical resistance (TEER), quantification of goblet and ciliated cells, real time PCR for MUC5AC, ELISA for VEGF, EGF and MCP-1 (CCL-2) and ELISA for MUC5AC were assessed.
We found that well-differentiated PBECs expressed IL-31-RA however it's expression did not increase upon stimulation with IL-31 or either of the other treatments. TEER indicated good formation of tight junctions which was found to be similar across all treatment groups (p=0.9). We found that IL-13 alone significantly reduced the number of ciliated cells compared with unstimulated (IL-13 stimuation: mean=4.8% (SD=2.5); unstimulated: mean=15.9%, (SD=7.4), p<0.01). IL-31 stimulation alone had no effect on ciliated cells whereas the IL-13/IL-31 combination stimulation significantly reduced the number of ciliated cells compared with control (IL-13/IL-31 combination: mean=5.1% (SD=4.6); unstimulated: mean=15.9%, (SD=7.4), p<0.01). We did not find that the combination of IL-13 and IL-31 had any additional effects to that of IL-13 alone. MUC5AC mRNA and secreted mucin was found in similar levels between unstimulated and all treatments, however IL-13 increased levels of MUC5AC mRNA by a factor of 2.84, albeit not significantly, compared with unstimulated cultures (IL-13 stimulation: mean=2.84 (SD=3.79); unstimulated: mean=1.0).
IL-31RA receptor is present on well-differentiated paediatric bronchial epithelial cells. IL-31 does not exhibit any detrimental effects on mucociliary differentiation. IL-31 does not appear to have a synergistic effect when combined in culture with IL-13, in the differentiation process.
白细胞介素-31（IL-31）是一种新型细胞因子，与特应性皮炎等过敏性疾病有关，最近还与哮喘有关。虽然IL-31在特应性皮炎等皮肤疾病中已得到充分研究，但关于IL-31在哮喘中的作用，特别是在过敏性哮喘发病机制核心的支气管上皮分化过程中的作用，人们了解甚少。
我们检测了白细胞介素-13（IL-13，20纳克/毫升）、白细胞介素-31（IL-31，20纳克/毫升）以及IL-13/IL-31联合刺激（各20纳克/毫升）对健康患者（n=6）的小儿支气管上皮细胞（PBECs）体外黏液纤毛分化的影响。评估了IL-31受体（IL-31-RA）的表达、分化标志物（杯状细胞和纤毛细胞）、跨上皮电阻（TEER）、杯状细胞和纤毛细胞的定量、MUC5AC的实时聚合酶链反应、血管内皮生长因子（VEGF）、表皮生长因子（EGF）和单核细胞趋化蛋白-1（MCP-1，CCL-2）的酶联免疫吸附测定（ELISA）以及MUC5AC的ELISA。
我们发现分化良好的PBECs表达IL-31-RA，但其表达在IL-31或其他任何处理刺激后并未增加。TEER表明紧密连接形成良好，在所有处理组中相似（p=0.9）。我们发现，与未刺激组相比，单独使用IL-13显著减少了纤毛细胞数量（IL-13刺激：平均值=4.8%（标准差=2.5）；未刺激：平均值=15.9%，（标准差=7.4），p<0.01）。单独使用IL-31刺激对纤毛细胞没有影响，而与对照组相比，IL-13/IL-31联合刺激显著减少了纤毛细胞数量（IL-13/IL-31联合：平均值=5.1%（标准差=4.6）；未刺激：平均值=15.9%，（标准差=7.4），p<0.
