Broide D H, Paine M M, Firestein G S
Department of Medicine, University of California, San Diego 92103.
J Clin Invest. 1992 Oct;90(4):1414-24. doi: 10.1172/JCI116008.
IL-5 and granulocyte macrophage-colony-stimulating factor (GM-CSF) are important regulators of eosinophil survival, proliferation, and effector function. To determine whether IL-5 and/or GM-CSF are generated by eosinophils at sites of allergic inflammation, we have used in situ hybridization with 35S-labeled RNA probes to study the expression of IL-5 and GM-CSF mRNA in bronchoalveolar lavage (BAL) eosinophils derived from asthmatics (n = 5) before and after endobronchial allergen challenge. Endobronchial allergen challenge induced a significant airway eosinophilia (pre-allergen challenge 0.6 +/- 0.5% eosinophilia vs post-allergen challenge 48.2 +/- 25.6% eosinophilia). Post-allergen challenge eosinophils expressed IL-5 and GM-CSF mRNA, but did not express IL-1 beta or IL-2 mRNA. To determine whether the IL-5 mRNA-positive cells coexpressed GM-CSF mRNA, double mRNA labeling experiments with a digoxigenin-11-UTP nonradioactive labeled IL-5 RNA probe and a GM-CSF 35S-labeled RNA probe were performed. These studies demonstrated that individual eosinophils expressed one of four cytokine mRNA profiles (IL-5+, GM-CSF+, 34 +/- 13%; IL-5+, GM-CSF-, 34 +/- 5%; IL-5-, GM-CSF+, 11 +/- 9%; IL-5-, GM-CSF-, 21 +/- 25%). The expression of IL-5 and GM-CSF by eosinophils at sites of allergic inflammation in asthmatics may provide an important autocrine pathway, maintaining the viability and effector function of the recruited eosinophils.
白细胞介素-5(IL-5)和粒细胞巨噬细胞集落刺激因子(GM-CSF)是嗜酸性粒细胞存活、增殖及效应功能的重要调节因子。为确定在变应性炎症部位嗜酸性粒细胞是否产生IL-5和/或GM-CSF,我们使用35S标记的RNA探针进行原位杂交,以研究支气管肺泡灌洗(BAL)嗜酸性粒细胞中IL-5和GM-CSF mRNA的表达,这些嗜酸性粒细胞来自5例哮喘患者,在支气管内过敏原激发前后获取。支气管内过敏原激发诱导了显著的气道嗜酸性粒细胞增多(过敏原激发前嗜酸性粒细胞增多为0.6±0.5%,而过敏原激发后为48.2±25.6%)。过敏原激发后的嗜酸性粒细胞表达IL-5和GM-CSF mRNA,但不表达IL-1β或IL-2 mRNA。为确定IL-5 mRNA阳性细胞是否共表达GM-CSF mRNA,进行了地高辛-11-UTP非放射性标记的IL-5 RNA探针和GM-CSF 35S标记的RNA探针的双mRNA标记实验。这些研究表明,单个嗜酸性粒细胞表达四种细胞因子mRNA谱之一(IL-5+、GM-CSF+,34±13%;IL-5+、GM-CSF-,34±5%;IL-5-、GM-CSF+,11±9%;IL-5-、GM-CSF-,21±25%)。哮喘患者变应性炎症部位嗜酸性粒细胞对IL-5和GM-CSF的表达可能提供一条重要的自分泌途径,维持募集的嗜酸性粒细胞的存活及效应功能。