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

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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.