Antibodies to tubulin and actin bind to the surface of a human monocytic cell line, U937

Intermittent reports of cytoskeleton proteins (actin and tubulin) on the cell surface have appeared over the last 13 years. Whereas most have concentrated on lymphocytes, this study provides evidence for the presence of these proteins on the surface of a human cultured monocyte-like cell line, U937....

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Bibliographic Details
Published in:The journal of histochemistry and cytochemistry 1991-07, Vol.39 (7), p.981-985
Main Authors: Por, SB, Cooley, MA, Breit, SN, Penny, R, French, PW
Format: Article
Language:English
Subjects:
Actins - analysis
Actins - immunology
Antibodies, Monoclonal
Biological and medical sciences
Cell Line
Cell structures and functions
Flow Cytometry
Fundamental and applied biological sciences. Psychology
Humans
Immunoblotting
Lipopolysaccharides - pharmacology
Microscopy, Electron, Scanning
Molecular and cellular biology
Monocytes - chemistry
Monocytes - physiology
Monocytes - ultrastructure
Tetradecanoylphorbol Acetate - pharmacology
Tubulin - analysis
Tubulin - immunology
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Summary:Intermittent reports of cytoskeleton proteins (actin and tubulin) on the cell surface have appeared over the last 13 years. Whereas most have concentrated on lymphocytes, this study provides evidence for the presence of these proteins on the surface of a human cultured monocyte-like cell line, U937. Both actin and tubulin were detected on the surface of U937 cells by flow cytometry, using an indirect staining procedure based on biotin-streptavidin-phycoerythrin, chosen for greater sensitivity. By use of this procedure, the majority of viable unstimulated U937 cells stained positively for actin and tubulin, although the level of fluorescence intensity was low. With an antibody specific for tyrosine-tubulin, most of the surface tubulin was also found to be tyrosinylated. For vimentin, an intermediate filament protein abundantly present in the cytoplasm of U937 cells, no staining could be detected. Confirmation of the flow cytometry data for surface actin and tubulin on unstimulated U937 cells was achieved by direct vesualization using a confocal laser scanning microscope. When U937 cells were activated with PMA and LPS, a marked reduction in the level of cell surface actin and tubulin occurred. The role of cell surface actin and tubulin on unstimulated U937 cells, in terms of monocyte function, remains to be elucidated.
ISSN:0022-1554
1551-5044
DOI:10.1177/39.7.1865114