Production of an allergic inflammation model using human lungs resected by a surgical operation and its application to drug discovery for the treatment of asthma

为了解决药理学中的物种差异问题，我们试图利用肺癌患者手术切除后的人肺组织制作炎症模型。用人IgE被动致敏的断片与抗人IgE抗体孵育后，与对照组(不加抗-IgE)相比，可产生大量的半胱氨酰白三烯(CysLTs)。单独添加人C3a或C5a并不能产生CysLT。然而，在抗IgE抗体中加入1.0 ng/ml的C5a进一步增强了CysLT的产生。因此，产生了一种以补体激活为病理生理机制的过敏性炎症模型。新的C5a受体补充肽或C5a受体拮抗剂均以剂量依赖的方式抑制1 ng/mlC5a引起的过敏反应所致的人肺碎片产生CysLT。…此外，由于过敏性毒素及其受体的一级结构存在可考虑的物种差异，这种使用人肺组织的模型可能对筛选新药有价值。相比之下，缺乏合适的人体组织，缺乏稳定保存的方法，阻碍了利用人体组织制作适当的模型。取肺癌患者的肺组织，在-5℃或4℃条件下保存5天，然后进行组织学和生化检查。-5℃保存的组织形态基本正常，支气管上皮纤毛完整，内皮细胞正常，4℃保存的组织结构退化。单链DNA是DNA断裂的标志，在4℃保存的组织中经常观察到，而在-5℃保存的组织中很少观察到。在5℃保存3天后，组织产生大量的细胞免疫反应，而保存4℃的组织对抗体刺激无反应。因此，这种新型的保存人体组织的过冷系统可能会促进人体组织在医学上的应用。较少

In order to resolve a problem concerning species differences in pharmacological science, we tried to make an inflammation model using human lung tissues obtained from patients with lung cancer after surgical resection. When the chopped lung fragments passively sensitized with human IgE were incubated with anti-human IgE antibody, a significant amount of cysteinyl-leukotrienes (cysLTs) was generated in comparison with the control (without anti-IgE). The addition of human C3a or C5a alone did not generate cysLTs. The co-addition, however, of C5a at doses of 1.0 ng/ml to the anti-IgE antibody further potentiated cysLT production. Consequently, an allergic inflammation model in which complement activation was involved as a pathophysiological mechanism was produced. Either a novel C5a receptor complementary peptide or a C5a receptor antagonist inhibited cysLT production in a dose-dependent manner by the human lung fragments following the anaphylactic reaction in the presence of 1 ng/ml C5a. … More Since there are consideable species differences in primary structure of anaphylatoxins and those receptors, this model using human lung tissues may be valuable to perform screening for a novel drug. In contrast, the shortage of suitable human tissues and lack of methods for stable preservation are hindering to produce an adequate model using human tissues. After lung tissues were obtained from patients with lung cancer, they were kept at -5℃ or 4℃ for as many as 5 days, and then they were histologically and biochemically examined. Although the tissues preserved at-5℃ had an almost normal morphology with intact cilia on bronchial epithelium and normal endothelium, the tissues stored at 4℃ showed degradation of these structures. Single-stranded DNA, a sign of DNA cleavage, was frequently noted in tissues stored at 4℃, but only rarely observed at -5℃. A significant amount of cysLTs was generated by anaphylactic reaction from tissues stored at 5℃ for 3 days, but there was no response to antibody stimulation from tissues at 4℃. Therefore, such the novel preservation of human tissues as super-cooling system may facilitate availability of human tissues in medical science. Less