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R Nozawa, H Kato, T Ito and T Yokota
Department of Bacteriology, Juntendo University School of Medicine, Hongo,
Tokyo.
Human promyelocytic leukemia (HL-60) cells were induced to differentiate
into macrophage-like cells by treatment with 10(-7) mol/L
1,25-dihydroxyvitamin D3 (VD3). A monoclonal antibody (MoAb, 60B8),
reactive with the particulate of the differentiated cells but not of the
untreated cells, was isolated. The antigen recognized by the MoAb became
apparent two days after VD3 treatment, and its concentration increased and
peaked on day 6. Human neutrophils, followed by monocytes and
differentiated HL-60 cells, showed the greatest abundance of the antigen.
Monocytes cultured for eight days in vitro lost the antigen. No 60B8
antigen was seen in other blood cells. The MoAb precipitated two
polypeptides with an apparent molecular weight (mol wt) of 15,000 (15 k)
and 13 k in the detergent-solubilized, 35S-methionine-labeled lysate of the
differentiated HL-60 cells. Double-sandwich type enzyme- linked
immunosorbent assay (ELISA) devised for the quantitative assay of 60B8
antigen indicated that some 2% to 5% of neutrophil protein was 60B8
antigen. This antigen was not exposed on the neutrophil cell surface, since
the cells were not stained immunofluorescently with either mono- or
polyclonal antibody, unless they had become permeable. The neutrophil
membrane and the granules were separated on the Percoll density gradient,
and the antigen was found localized in the plasma membrane-rich fraction.
These findings suggested that 60B8 antigen is a novel differentiation
antigen for phagocytic cells.
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| Copyright © 1988 by American Society of Hematology Online ISSN: 1528-0020 | |||||||||