A monoclonal antibody against human neutrophil: evaluation of its effects on neutrophil function and partial characterization of the antigen
Ozaki, Y.; Ohashi, T.; Ohto, H.; Maeda, H.
Nihon Ketsueki Gakkai Zasshi Journal of Japan Haematological Society 48(4): 980-991
1985
ISSN/ISBN: 0001-5806 PMID: 3904295 Document Number: 254611
A hybridoma technique was used to produce a monoclonal antibody against human neutrophils. The IgM class antibody (TG-8) was enriched in ascites at a concentration of 900 mg/dl. The reactivity of TG-8 with blood cells and cultured cell lines suggested that it recognized a differentiation antigen which appears between the myeloblast and myelocyte stages. TG-8 immunoprecipitated polypeptides of 145 k daltons, 125 k daltons, and 92 k daltons from 125I-labeled neutrophil membrane proteins. It also immunoprecipitated a major band of 120-150 k daltons and a minor band of 92 k daltons from 3H-labeled neutrophil membrane glycoproteins, as analyzed by NaDodSO4 polyacrylamide gel electrophoresis. In functional studies, 1/1000-1/100 diluted TG-8 induced myeloperoxidase release in the presence of cytochalasin B. It also enhanced the random migration of nuetrophil chemotaxis. Significant aggregation of neutrophils was induced by TG-8, though this phenomenon was partially due to cell to cell cross-linkage by IgM molecules. TG-8 had no effects on yeast uptake, zymosan-induced myeloperoxidase release, fMLP-induced true chemotaxis, or chemiluminescence.