Till S, Li B, Durham S, Humbert M, Assoufi B, Huston D, Dickason R, Jeannin P, Kay A B, Corrigan C
Department of Allergy and Clinical Immunology, National Heart and Lung Institute, London, GB.
Eur J Immunol. 1995 Oct;25(10):2727-31. doi: 10.1002/eji.1830251002.
Specific eosinophil accumulation and activation within the asthmatic bronchial mucosa are thought to occur at least partly through the actions of cytokines, including interleukin (IL)-5, IL-3 and granulocyte/macrophage colony-stimulating factor (GM-CSF). Although mRNA encoding some of these cytokines has been demonstrated in bronchoalveolar lavage (BAL) fluid cells and bronchial biopsies from asthmatics, it has yet to be established whether these cells produce the translated products and whether expression is associated with CD4+ T helper or CD8+ cytotoxic T cells. We addressed this problem by raising polyclonal CD4+ and CD8+ T cell lines from the BAL fluid of six atopic asthmatics, five atopic non-asthmatics and seven non-atopic non-asthmatic controls. BAL fluid cells obtained at fiberoptic bronchoscopy were depleted of adherent cells, and then T lymphocytes expanded by stimulation with monoclonal anti-CD3 antibody and recombinant human IL-2. When lymphocytes had expanded to sufficient numbers, CD4+ and CD8+ cells were separated by positive selection with magnetic beads coated with anti-CD4 or anti-CD8 monoclonal antibodies and further expanded. Cytokine secretion by standardized cell numbers was measured by enzyme-linked immunosorbent assays. BAL CD4+ T cell lines from the asthmatics secreted significantly elevated quantities of both IL-5 and GM-CSF as compared with lines from the atopic and non-atopic controls (p = 0.023-0.003). In contrast, IL-3 secretion did not significantly differ between the groups. In some subjects, CD8+ T cell lines also secreted significant quantities of these cytokines and there was a trend for IL-5 secretion by these cells to be higher in asthmatics than non-atopic controls (p = 0.035). These data are consistent with the hypothesis that activated T lymphocytes from asthmatics, particularly of the CD4+ subset, are predisposed to release elevated quantities of cytokines relevant to the accumulation and activation of eosinophils.
哮喘支气管黏膜内特定的嗜酸性粒细胞聚集和活化被认为至少部分是通过细胞因子的作用发生的，这些细胞因子包括白细胞介素（IL）-5、IL-3和粒细胞/巨噬细胞集落刺激因子（GM-CSF）。尽管在哮喘患者的支气管肺泡灌洗（BAL）液细胞和支气管活检组织中已证实存在编码其中一些细胞因子的mRNA，但这些细胞是否产生翻译产物以及表达是否与CD4⁺辅助性T细胞或CD8⁺细胞毒性T细胞相关仍有待确定。我们通过从6名特应性哮喘患者、5名特应性非哮喘患者和7名非特应性非哮喘对照者的BAL液中培养多克隆CD4⁺和CD8⁺T细胞系来解决这个问题。在纤维支气管镜检查时获得的BAL液细胞去除贴壁细胞，然后用单克隆抗CD3抗体和重组人IL-2刺激使T淋巴细胞扩增。当淋巴细胞扩增到足够数量时，用包被抗CD4或抗CD8单克隆抗体的磁珠进行阳性选择分离CD4⁺和CD8⁺细胞并进一步扩增。通过酶联免疫吸附测定法测量标准化细胞数量的细胞因子分泌。与特应性和非特应性对照者的细胞系相比，哮喘患者的BAL CD4⁺T细胞系分泌的IL-5和GM-CSF量显著升高（p = 0.023 - 0.003）。相比之下，各组间IL-3分泌无显著差异。在一些受试者中，CD8⁺T细胞系也分泌大量这些细胞因子，并且哮喘患者中这些细胞分泌IL-5的趋势高于非特应性对照者（p = 0.035）。这些数据与以下假设一致，即哮喘患者的活化T淋巴细胞，特别是CD4⁺亚群，易于释放与嗜酸性粒细胞聚集和活化相关的细胞因子的量增加。
