Molecular action mechanism of eosinophil granule proteins in destruction of mucosal cells in ulcerative colitis

为了探讨活化的嗜酸性粒细胞在溃疡性结肠炎炎症过程中的作用，我们制备了抗人嗜酸性粒细胞颗粒蛋白多肽的抗体，试图纯化ECP1。根据Barker等人报道的人ECP氨基酸序列，选择了以下四个ECP多肽：ECP1，CRYADRPGRRFVVA；ECP2，CDNRDPRDSPRYPVV；ECP3，CTIAMRAINNYRWRC；和ECP4，TRAQWFAIQHISLNPPR.2。用ECP多肽抗原免疫兔，获得了即使将兔抗血清稀释到1/10000也能与合成肽反应的抗人ECP抗体。3.所获得的抗人ECP抗体在Pharmacia ECP RIA试剂盒的ECP检测系统中均不能反应为抗ECP抗血清。4.抗人ECP2抗体和抗人ECP3抗体与嗜酸性粒细胞和单核细胞免疫组织化学染色均呈阳性反应。上述结果表明，虽然需要确定获得的抗ECP抗体是否与人ECP发生反应，但至少ECP2和ECP3氨基酸序列区域具有免疫反应性。抗人ECP抗体的抗炎作用应通过实验动物模型进行研究。

To investigate the role of activated eosinophils in the inflammatory process of ulcerative colitis, we prepared antibodies against polypeptides of a human eosinophil granule protein, eosinophil cationic protein (ECP), in attempt to purify ECP.1. The following four ECP polypeptides were selected based on the human ECP amino acid sequence reported by Barker et al.: ECP1, CRYADRPGRRFVVA; ECP2, CDNRDPRDSPRYPVV; ECP3, CTIAMRAINNYRWRC; and ECP4, TRAQWFAIQHISLNPPR.2. Rabbits were immunized with ECP polypeptide antigens, and anti-human ECP antibodies that reacted with synthetic peptides even the rabbit antisera were diluted to 1/10,000 were obtained. 3. None of the acquired anti-human ECP antibodies reacted as anti-ECP antiserum in an ECP assay system using Pharmacia ECP RIA kit. 4. Anti-human ECP2 antibody and anti-human ECP3 antibody reacted with eosinophils and monocytes in immunohistochemical staining.The above findings suggested that although it is necessary to confirm whether the acquired anti-ECP antibodies react with human ECP, at least the ECP2 and ECP3 amino acid sequence regions have immunological reactivity. The anti-inflammatory effect of the anti-human ECP antibodies should be investigated using an experimental animal model.