BackgroundFarm‐dust mediated asthma protection in childhood was replicated in numerous epidemiological studies. Central immune mechanisms are not fully understood. This exploratory study aimed to disentangle underlying immunological regulation of farm‐dust mediated protection in peripheral blood on a single‐cell level.MethodsSingle‐cell protein expression of in vitro farm‐dust stimulated and unstimulated cells from allergic asthmatics and healthy controls were measured using mass cytometry. Analysis of innate and adaptive cellular proportions (linear regression) and T‐cell proliferation was performed. Functional marker intensity was investigated using Earth Mover's Distance and the Monte Carlo permutation test.ResultsFarm‐dust stimulation induced cell type‐specific regulation: Key‐features of farm‐dust stimulation comprised opposing regulation of immune‐cell frequencies (downregulated innate cell populations (monocytes/DCs (p < .001), NK‐cells (p < .05)) and upregulated adaptive populations (B‐cells, CD4+ T‐cells (both p < .05)), reduced CD4+ CD25− T‐cell proliferation, and differential cell type‐specific functional marker expression. Following stimulation, functional marker analysis revealed induced activation (CD25) in T‐cells and NK‐T‐cells in both phenotypes even after correction for multiple testing. Cytotoxicity (GZMB) and inflammation (pERK1/2, pp38) related markers were reduced in T‐cells exclusively in asthmatic children. Asthma‐associated markers (Gata3, RORγ, and HLA‐DR) were reduced in T‐ and innate‐ cell populations of asthmatics following stimulation. B‐cells displayed a phenotypically independent increase of diverse functional markers upon farm‐dust stimulation.ConclusionsThis study mimicking in vivo environmental exposure identified a novel profile of immune‐regulatory markers using mass cytometry demonstrating decreased asthma‐associated markers following farm‐dust stimulation. These findings may be key for further studies on asthma prevention in childhood.

中文翻译：

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农场粉尘介导的儿童哮喘保护：质谱流式细胞术揭示了新的细胞调节


背景农场粉尘介导的儿童哮喘保护作用在许多流行病学研究中得到复制。中枢免疫机制尚不完全清楚。这项探索性研究旨在在单细胞水平上解开农场粉尘介导的外周血保护的潜在免疫调节。方法使用质谱流式细胞术测量来自过敏性哮喘患者和健康对照者的体外农场粉尘刺激和未刺激细胞的单细胞蛋白表达。分析先天性和适应性细胞比例 （线性回归） 和 T 细胞增殖。使用地球移动器距离和蒙特卡洛排列检验研究功能标志物强度。结果农场粉尘刺激诱导的细胞类型特异性调节：农场粉尘刺激的主要特征包括免疫细胞频率的相反调节（下调的先天细胞群 （单核细胞/DC （p < .001），NK 细胞 （p < .05）））和上调的适应性群体 （B 细胞，CD4 + T 细胞 （均为 p < .05）），CD4 + CD25 - T 细胞增殖减少，以及差异细胞类型特异性功能标志物表达。刺激后，功能标志物分析显示 T 细胞和 NK-T 细胞在两种表型中均诱导激活 （CD25），即使在多次测试校正后也是如此。仅在哮喘儿童的 T 细胞中，细胞毒性 （GZMB） 和炎症 （pERK1/2，pp38） 相关标志物降低。刺激后，哮喘患者的 T 细胞群和先天细胞群中的哮喘相关标志物 （Gata3 、 RORγ 和 HLA-DR） 减少。B 细胞在农场粉尘刺激后表现出不同功能标志物的表型独立增加。结论这项模拟体内环境暴露的研究使用质谱流式细胞术确定了免疫调节标志物的新特征，表明农场粉尘刺激后哮喘相关标志物减少。这些发现可能是进一步研究儿童哮喘预防的关键。