Summary
We have characterized the cellular composition of preparations isolated from peripheral blood by Ficoll-Isopaque gradient centrifugation.125I-insulin binding to every cell type was measured. A highly significantly positive correlation between specific cell binding fraction and the monocyte concentration of the heterogeneous cell suspension was demonstrated. Depletion of monocytes reduced the insulin binding approximately 80%, which confirms previous findings by other investigators. The granulocytes possessed the second highest binding ability, but only one fourteenth of that of monocytes. Compared to the lymphocyte the monocyte had about 25 times greater insulin binding. Also thrombocytes bound insulin and contamination with these meant that their contribution to the total specific cell binding was not negligible. A reduction in these contaminants is essential. We found that insulin binding to erythrocytes was insignificant. A method of calculating the specific insulin binding to monocytes alone is introduced. The monocyte-insulin-receptor possesses specificity. Only an insignificant degradation of receptor bound insulin could be shown. Evidence of negative cooperativity between receptors was found. Consequently monocytes are considered a useful model for insulin receptor studies in man.
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Gavin, J.R. III, Roth, J., Jen, P., Freychet, P.: Insulin receptors in human circulating cells and fibroblasts. Proc. Natl. Acad. Sci. USA69, 747–751 (1972)
Krug, U., Krug, F., Cuatrecasas, P.: Emergence of insulin receptors on human lymphocytes during in vitro transformation. Proc. Natl. Acad. Sci. USA69, 2604–2608 (1972)
Schwartz, R.H., Bianco, A.R., Handwerger, B.S., Kahn, R.C.: Demonstration that monocytes rather than lymphocytes are the insulin binding cells in preparations of human peripheral blood mononuclear leucocytes: implications for studies of insulin-resistant states in man. Proc. Natl. Acad. Sci. USA72, 474–478 (1975)
Boyum, A.: Separation of leucocytes from blood and bone marrow. Scand. J. Clin. Lab. Invest.21 (supp. 197), 77–89 (1968)
Pedersen, O., Beck-Nielsen, H.: A study of insulin receptors in human mononuclear leucocytes. Acta Endocrinol. (Kbh)83, 556–564 (1976)
Yam, L.T., Li, C.Y., Crosby, W.N.: Cytochemical identification of monocytes and granulocytes. Am. J. Clin. Pathol.55, 283–290 (1971)
Stjernsvärdt, J., Tondal, M., Vanky, F., Wigzell, H., Sealy, R.: Lymphopenia and change in distribution of human B and T lymphocytes in peripheral blood induced by irradiation for mammary carcinoma. Lancet1972 I, 1352–2356
Greenwalt, T., Gatewski, M., McKenna, J.: A new method for preparing buffy coat-poor blood. Transfusion2, 221–225 (1962)
Boyum, A.: Separation of blood leucocytes, granulocytes and lymphocytes. Tissue Antigens4, 269–274 (1974)
Svejgård, A., Kissmeyer-Nielsen, F.: Complement-fixing platlet iso-antibodies. I. A quantitative technique for their detection. Vox Sang.14, 106–118 (1968)
Andreasen, P.A., Schaumburg, P., Østerlind, K., Vinten, J., Gammeltoft, S., Gliemann, J.: A rapid technique for separation of thymocytes from suspensions by centrifugation through silicone oil. Anal. Biochem.59, 110–115 (1974)
Rodbell, M., Michiel, H., Krans, J., Pohl, S., Birnhammer, L.: The glucagon-sensitive adenyl cyclase system in plasma membranes of rat liver. III. Binding of glucagon: Method of assay and specificity. J. Biol. Chem.246, 1861–1871 (1971)
Scatchard, G.: The attraction of proteins for small molecules and ions. Ann. N. Y. Acad. Sci.51, 660–672 (1949)
De Meyts, P. and Roth, J.: Cooperativity in ligand binding: A New graphic analysis. Biochem. Biophys. Res. Commun.66, 1118–1125 (1975)
De Meyts, P., Bianco, A.R., Roth, J.: Site-site interactions among insulin receptors. J. Biol. Chem.251, 1877–1888 (1976)
Fussganger, R.D., Kahn, R.C., Roth, J., De Meyts, P.: Binding and degradation of insulin by human peripheral granulocytes. J. Biol. Chem.251, 2761–2769 (1976)
Gavin, J.R. III, Gordon, P., Roth, J., Archer, J.A., Buell, D.N.: Characteristics of the human lymphocyte insulin receptor, J. Biol. Chem.248, 2202–2207 (1973)
Terris, S., Steiner, D.F.: Binding and degradation of125I-insulin by rat hepatocytes. J. Biol. Chem.250, 8389–8398 (1975)
Bar, R., Gordon, P., Roth, J., Kahn, C.R., De Meyts, P.: Fluctuations in the affinity and concentration of insulin receptors on circulating monocytes of obese patients. J. Clin. Invest.58, 1123–1135 (1976)
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Beck-Nielsen, H., Pedersen, O., Kragballe, K. et al. The monocyte as a model for the study of insulin receptors in man. Diabetologia 13, 563–569 (1977). https://doi.org/10.1007/BF01236308
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DOI: https://doi.org/10.1007/BF01236308