| Research Abstract |
N-terminal peptides of human erythrocyte calpain (calcium dependent cysteine proteinase) were synthesized and their chemotactic and degranulation activities for human peripheral leukocytes were measured. N-acetyl Met-Phe-Leu-Val-Asn-Ser-Phe-Leu-Lys-Gly-Gly-Gly-Gly, N-acety Met-Phe-Leu-Val-Asn-Ser-Phe-Leu-Lys, N-acety Met-Phe-Leu-Val-Asn-Ser-Phe-Leu, and N-acety Met-Phe-Leu-Val-Asn-Ser-Phe had potent chemotactic activity. The smallest chemotactic factor was N-acetyl Met-Phe-Leu-Val. Moreover, these peptides were found to have potent chemotactic activity for human peripheral monocytes and cultured leukemia cell lines, THP-1, U-937, BALL and MT-1. The optimum concentration of chemotactic factor for these cells was 100 pM. Not only N-acetyl peptides but N-formyl peptides also had chemotactic and degranulation activities. Most of the N-formyl peptides showed higher chemotactic and degranulation activities than the corresponding N-acetyl peptides for neutrophils. Neutrophils from 42 healthy
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volunteers, 18 patients with psoriasis valgaris, 5 patients with pustulosis palmaris et plantaris, 4 patients with tonsillar focal infection and a patient with rheumatoid arthritis were measured for degranulation activity with N-formyl and N-acetyl peptides. The degranulation activity from these patient neutrophils was 2 times higher than that of healthy persons. On the other hand, in vivo experiment was carried out with these peptides. N-acetyl peptides were injected into rabbits through ear vein. After 90min, rabbit peripheral neutrophils increased about 30%. N-acetyl Met-Phe-Leu-Val was intracutaneously injected into guinea pig ear with or without bradykinin and neutrophil infiltration in dermis and subcutis was examined. Intensive neutrophil accumulation was observed 2 hr after the injection of chemotactic peptide and bradykinin indicating the cooperative effect of bradykinin with chemotactic factor. Preparation of monoclonal antibodies against calpain N-terminal peptides was attempted and a cell line produced monoclonal antibody against N-acetyl Ser-Glu-Glu-Ile-Ile-Thr-Pro-Val-Tyr could be obtained, but monoclonal antibodies against other peptides were not produced, perhaps because of weak antigenecity. Establishment of monoclonal antibodies against other peptides are now under preparation using peptides consisting of more than 20 amino acid residues as antigen. Less
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