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Receptor binding and internalization of a water-soluble (1-->3)-beta-D-glucan biologic response modifier in two monocyte/macrophage cell lines.

A Müller, P J Rice, H E Ensley, P S Coogan, J H Kalbfleish, J L Kelley, E J Love, C A Portera, T Ha, I W Browder and D L Williams
J Immunol May 1, 1996, 156 (9) 3418-3425;
A Müller
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P J Rice
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H E Ensley
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P S Coogan
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J H Kalbfleish
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J L Kelley
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E J Love
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C A Portera
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T Ha
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I W Browder
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D L Williams
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Abstract

Glucan phosphate, a water-soluble, chemically defined (1-->3)-beta-D-glucan biologic response modifier, has been reported to exert antisepsis activity and accelerate wound healing. In this study we describe the specific binding of glucan phosphate to human and murine monocyte/macrophage cell lines, U937 and J774A.1, respectively. At 37 degrees C, equilibrium binding was rapidly achieved, i.e., within 1 min. In U937 cells, binding occurred with an affinity (Kd) of 37 microM and a Bmax of 65 x 106 binding sites/cell at 37 degrees C. In J774A.1 cells, glucan phosphate bound with an affinity (Kd) of 24 microM and a Bmax of 53 x 106 binding sites/cell at 37 degrees C. In both cases there was insignificant nonspecific binding. We further demonstrated that bound glucan phosphate cannot be displaced by a 50-fold excess of unlabeled ligand, suggesting internalization of glucan phosphate. Transmission electron microscopy showed significantly increased cytoplasmic vacuolization and significantly decreased mitotic activity in glucan phosphate-treated U937 cells compared with that in untreated cells. Pullulan, a random coil alpha-(1-->4)-(1-->6)-linked glucose polymer that served as a control, did not compete for the same binding site as glucan phosphate in either cell line, indicating the specificity of the binding site for (1-->3)-beta-D-glucans. We conclude that water-soluble pharmaceutical grade (1-->3)-beta-D-glucan phosphate specifically binds to and is internalized by U937 and J774A.1 cells.

  • Copyright © 1996 by American Association of Immunologists

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The Journal of Immunology
Vol. 156, Issue 9
1 May 1996
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Receptor binding and internalization of a water-soluble (1-->3)-beta-D-glucan biologic response modifier in two monocyte/macrophage cell lines.
A Müller, P J Rice, H E Ensley, P S Coogan, J H Kalbfleish, J L Kelley, E J Love, C A Portera, T Ha, I W Browder, D L Williams
The Journal of Immunology May 1, 1996, 156 (9) 3418-3425;

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Receptor binding and internalization of a water-soluble (1-->3)-beta-D-glucan biologic response modifier in two monocyte/macrophage cell lines.
A Müller, P J Rice, H E Ensley, P S Coogan, J H Kalbfleish, J L Kelley, E J Love, C A Portera, T Ha, I W Browder, D L Williams
The Journal of Immunology May 1, 1996, 156 (9) 3418-3425;
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Print ISSN 0022-1767        Online ISSN 1550-6606