Wosinska-Becler K, Plewako H, Håkansson L, Rak S
Asthma and Allergy Research Group, Department of Respiratory Medicine and Allergology, Sahlgrenska University Hospital, Göteborg, Sweden.
Clin Exp Allergy. 2004 Jan;34(1):123-30. doi: 10.1111/j.1365-2222.2004.01850.x.
The naturally occurring pollen season permits observation of the kinetic changes in the process of allergic inflammation. We examined cytokine production in peripheral blood (PB) T cells and monocytes obtained from birch-allergic patients both during and outside the pollen season.
PB from 16 patients and six healthy controls was obtained during the alder pollen season, at the beginning and the peak of the birch pollen season and outside the pollen season. Mononuclear cells (MNC) were stimulated with allergen and polyclonal activators. For flow cytometric analysis, MNC were stained with monoclonal antibodies (MoAbs) against the cell surface markers CD3, CD8, CD14 and the intracellular cytokines IL-4, IL-5, IL-10, IL-12, IL-13, granulocyte macrophage-colony stimulating factor (GM-CSF) and IFN-gamma.
In allergic patients, significant increases in clinical symptoms, use of medication, eosinophil numbers and birch-specific IgE were found during the pollen season. In vitro allergen stimulation increased the number of GM-CSF+ monocytes (P<0.01) and this increase was dependent on allergen exposure. The IL-4/IFN-gamma ratio rose (P<0.001) at the peak of birch pollen season and the ratio correlated with symptom scores during the birch season. In the CD4+ cell population, the numbers of GM-CSF+ cells were higher throughout the alder and birch seasons compared with outside the pollen season (P<0.05). No such changes were seen in the healthy controls.
The main finding of our study was the increased percentage of GM-CSF+ monocytes in atopic subjects compared with healthy controls. In allergic patients, natural seasonal pollen exposure resulted in increased numbers of GM-CSF+ cells among both monocytes and CD4+ T cells. We have also shown that a seasonal change in Th2/Th1 cytokine ratio requires an adequate and prolonged allergen stimulation that is seen late in the pollen season.
自然出现的花粉季节有助于观察变应性炎症过程中的动力学变化。我们检测了桦树过敏患者在花粉季节期间及非花粉季节外周血（PB）T细胞和单核细胞中细胞因子的产生情况。
在桤木花粉季节、桦树花粉季节开始时、高峰时以及非花粉季节采集了16例患者和6例健康对照者的PB。用变应原和多克隆激活剂刺激单核细胞（MNC）。对于流式细胞术分析，用针对细胞表面标志物CD3、CD8、CD14以及细胞内细胞因子白细胞介素-4（IL-4）、白细胞介素-5（IL-5）、白细胞介素-10（IL-10）、白细胞介素-12（IL-12）、白细胞介素-13（IL-13）、粒细胞巨噬细胞集落刺激因子（GM-CSF）和干扰素-γ（IFN-γ）的单克隆抗体（MoAb）对MNC进行染色。
在过敏患者中，花粉季节期间临床症状、药物使用、嗜酸性粒细胞数量和桦树特异性IgE显著增加。体外变应原刺激增加了GM-CSF+单核细胞的数量（P<0.01），且这种增加依赖于变应原暴露。在桦树花粉季节高峰时，IL-4/IFN-γ比值升高（P<0.001），且该比值与桦树季节期间的症状评分相关。在CD4+细胞群体中，与非花粉季节相比，在整个桤木和桦树季节GM-CSF+细胞数量更高（P<0.05）。健康对照者未观察到此类变化。
我们研究的主要发现是与健康对照者相比，特应性受试者中GM-CSF+单核细胞的百分比增加。在过敏患者中，自然季节性花粉暴露导致单核细胞和CD4+T细胞中GM-CSF+细胞数量增加。我们还表明，Th2/Th1细胞因子比值的季节性变化需要在花粉季节后期出现的充分且持续时间较长的变应原刺激。
