Atsuta J, Fujisawa T, Iguchi K, Terada A, Kamiya H, Sakurai M
Department of Pediatrics, Mie National Hospital, Japan.
Int Arch Allergy Immunol. 1995;108 Suppl 1:31-5. doi: 10.1159/000237197.
The effects of transforming growth factor (TGF) beta 1 on cytokine-enhanced eosinophil survival and degranulation were investigated in vitro to determine whether it is an inhibitory regulator of allergic inflammation. Peripheral blood eosinophils purified by Percoll density gradient centrifugation and the CD16 negative selection technique were incubated in the presence of eosinophil-activating cytokines (interleukin-5 (IL-5), IL-3, granulocyte-macrophage colony-stimulating factor (GM-CSF), interferon (IFN)-gamma) with and without TFG-beta 1 for 1-3 days. On day 1, eosinophil protein X release was measured by radioimmunoassay. Eosinophil viability on day 3 was determined by staining the cells with fluorescein diacetate and propidium iodide, and on the same day DNA was extracted and subjected to gel electrophoresis to test for fragmentation. TGF-beta 1 significantly inhibited eosinophil survival enhanced by IL-5, IL-3, GM-CSF and IFN-gamma in a dose-dependent manner. The inhibitory effects of TGF-beta 1 on IL-5-enhanced survival was partially reversed by high concentrations of IL-5 and was completely neutralized with anti-TGF-beta antibody. IL-5 inhibited DNA fragmentation of eosinophils in vitro. TGF-beta reversed the effect of IL-5, indicating that TGF-beta 1 activates the pathway of apoptosis. TGF-beta 1 significantly suppressed eosinophil protein X release induced by IL-5. These results suggest that TGF-beta 1 may play a role in the modulation of allergic inflammation.
为了确定转化生长因子（TGF）β1是否为变应性炎症的抑制性调节因子，我们在体外研究了其对细胞因子增强的嗜酸性粒细胞存活和脱颗粒的影响。通过Percoll密度梯度离心和CD16阴性选择技术纯化的外周血嗜酸性粒细胞，在有或无TFG-β1的情况下，与嗜酸性粒细胞激活细胞因子（白细胞介素-5（IL-5）、IL-3、粒细胞-巨噬细胞集落刺激因子（GM-CSF）、干扰素（IFN）-γ）一起孵育1至3天。在第1天，通过放射免疫测定法测量嗜酸性粒细胞蛋白X的释放。在第3天，通过用荧光素二乙酸酯和碘化丙啶对细胞进行染色来确定嗜酸性粒细胞的活力，并且在同一天提取DNA并进行凝胶电泳以检测片段化。TGF-β1以剂量依赖的方式显著抑制由IL-5、IL-3、GM-CSF和IFN-γ增强的嗜酸性粒细胞存活。TGF-β1对IL-5增强的存活的抑制作用被高浓度的IL-5部分逆转，并用抗TGF-β抗体完全中和。IL-5在体外抑制嗜酸性粒细胞的DNA片段化。TGF-β逆转了IL-5的作用，表明TGF-β1激活了凋亡途径。TGF-β1显著抑制由IL-5诱导的嗜酸性粒细胞蛋白X的释放。这些结果表明，TGF-β1可能在变应性炎症的调节中发挥作用。
