scholarly journals Inhibition of protein N-glycosylation by 2-deoxy-2-fluoro-d-galactose

1992 ◽  
Vol 285 (3) ◽  
pp. 821-826 ◽  
Author(s):  
V Gross ◽  
W E Hull ◽  
U Berger ◽  
T Andus ◽  
W Kreisel ◽  
...  
Keyword(s):  
Interleukin 6 ◽  
Ex Vivo ◽  
Primary Cultures ◽  
Blood Monocytes ◽  
Galactose Metabolism ◽  
Rat Hepatocyte ◽  
Peripheral Blood Monocytes ◽  
Efficient Inhibitor ◽  
Acid Glycoprotein ◽  
Alpha 1 Antitrypsin

The effects of 2-deoxy-2-fluoro-D-galactose (dGalF) on N- and O-glycosylation of proteins was studied in rat hepatocyte primary cultures and in human monocytes. In hepatocytes, dGalF at concentrations of 1 mM or higher completely inhibited N-glycosylation of alpha 1-antitrypsin and alpha 1-acid glycoprotein, whereas 4 mM-2-deoxy-D-galactose (dGal) only slightly impaired N-glycosylation. In monocytes, 1 mM- or 4 mM-dGalF blocked N-glycosylation of alpha 1-antitrypsin and of interleukin-6, while O-glycosylation of interleukin-6 remained unaffected. In monocytes, dGal had no effect on protein N-glycosylation. Addition of uridine effectively prevented the UTP deficiency induced by dGalF, but had no effect on the inhibition of protein N-glycosylation by dGalF. Using 19F-n.m.r. spectroscopy, 2-deoxy-2-fluoro-D-galactose 1-phosphate (dGalF-1-P), UDP-dGalF and UDP-dGlcF could be identified as the major metabolites of dGalF in hepatocytes as well as in monocytes. In conclusion, compared with dGal, dGalF is a more efficient inhibitor of protein N-glycosylation. The effect is not caused by the depletion of UTP induced by dGalF, but rather by metabolites of dGalF. dGalF is metabolized not only in hepatocytes but also in peripheral blood monocytes, which can be used for ex vivo studies of disturbances in D-galactose metabolism.

Effect of Steroid Administration Ex Vivo on the IκB/NF-κB Pathway in Human Peripheral Blood Monocytes

2003 ◽  
Vol 54 (5) ◽  
pp. 542
Author(s):  
Ho Il Yoon ◽  
Hee Seok Lee ◽  
Chang Hoon Lee ◽  
Choon Taek Lee ◽  
Young Whan Kim ◽  
...  
Keyword(s):  
Peripheral Blood ◽  
Ex Vivo ◽  
Human Peripheral Blood ◽  
Blood Monocytes ◽  
Peripheral Blood Monocytes ◽  
Steroid Administration

scholarly journals Interleukin-1 induces interleukin-6 production in peripheral blood monocytes

Blood ◽  
1990 ◽  
Vol 75 (6) ◽  
pp. 1305-1310 ◽  
Author(s):  
G Tosato ◽  
KD Jones
Keyword(s):  
Endothelial Cells ◽  
Interleukin 6 ◽  
Peripheral Blood ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Relative Molecular Mass ◽  
Blood Monocytes ◽  
Peripheral Blood Monocytes ◽  
Potent Inducer ◽  
Size Heterogeneity

Abstract Interleukin-6 (IL-6), a multifunctional cytokine produced in monocytes, fibroblasts, endothelial cells, and keratinocytes, is induced by a variety of stimulating signals, including lipopolysaccharide (LPS), poly (I), poly (C), IL-1, tumor necrosis factor (TNF), and platelet- derived growth factor. Some of these signals induce IL-6 effectively only in one cell type, and this selectivity of induction may explain selectivity of biologic effects. In the present study, we show that IL- 1 beta, previously known to be a potent inducer of IL-6 in fibroblasts, endothelial cells, and keratinocytes, but not in monocytes, is also a potent inducer of IL-6 in peripheral blood monocytes. High level IL-6 activity that could be neutralized by specific antibodies to IL-6 was detected in supernatants of IL-1-stimulated monocytes. Maximal induction required IL-1 concentrations of 10 ng/mL. As judged by sodium dodecyl sulfate-polyacrylamide gel electrophoresis under reducing conditions, IL-6 species of relative molecular mass of 19 to 26 Kd could be specifically immunoprecipitated from supernatants of IL-1- induced monocytes. Size heterogeneity is a reported feature of IL-6 produced in a variety of cell types, and monocyte-derived IL-6 induced by either IL-1 or LPS displayed similar size heterogeneity. The highly purified recombinant IL-1 beta preparation used contained little, if any, LPS. In addition, monocyte production of IL-6, induced by IL-1 beta, was specifically neutralized by anti-IL-1 beta antibodies, demonstrating that IL-1, rather than a contaminant in the IL-1 preparation, was responsible for IL-6 induction. A number of biologic activities have been ascribed both to IL-1 and IL-6. The finding that IL-1 induced IL-6 in monocytes may help in defining the spectrum of biologic activities of each of these interactive cytokines.

scholarly journals Interleukin-1 induces interleukin-6 production in peripheral blood monocytes

Blood ◽  
1990 ◽  
Vol 75 (6) ◽  
pp. 1305-1310 ◽  
Author(s):  
G Tosato ◽  
KD Jones
Keyword(s):  
Endothelial Cells ◽  
Interleukin 6 ◽  
Peripheral Blood ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Relative Molecular Mass ◽  
Blood Monocytes ◽  
Peripheral Blood Monocytes ◽  
Potent Inducer ◽  
Size Heterogeneity

Interleukin-6 (IL-6), a multifunctional cytokine produced in monocytes, fibroblasts, endothelial cells, and keratinocytes, is induced by a variety of stimulating signals, including lipopolysaccharide (LPS), poly (I), poly (C), IL-1, tumor necrosis factor (TNF), and platelet- derived growth factor. Some of these signals induce IL-6 effectively only in one cell type, and this selectivity of induction may explain selectivity of biologic effects. In the present study, we show that IL- 1 beta, previously known to be a potent inducer of IL-6 in fibroblasts, endothelial cells, and keratinocytes, but not in monocytes, is also a potent inducer of IL-6 in peripheral blood monocytes. High level IL-6 activity that could be neutralized by specific antibodies to IL-6 was detected in supernatants of IL-1-stimulated monocytes. Maximal induction required IL-1 concentrations of 10 ng/mL. As judged by sodium dodecyl sulfate-polyacrylamide gel electrophoresis under reducing conditions, IL-6 species of relative molecular mass of 19 to 26 Kd could be specifically immunoprecipitated from supernatants of IL-1- induced monocytes. Size heterogeneity is a reported feature of IL-6 produced in a variety of cell types, and monocyte-derived IL-6 induced by either IL-1 or LPS displayed similar size heterogeneity. The highly purified recombinant IL-1 beta preparation used contained little, if any, LPS. In addition, monocyte production of IL-6, induced by IL-1 beta, was specifically neutralized by anti-IL-1 beta antibodies, demonstrating that IL-1, rather than a contaminant in the IL-1 preparation, was responsible for IL-6 induction. A number of biologic activities have been ascribed both to IL-1 and IL-6. The finding that IL-1 induced IL-6 in monocytes may help in defining the spectrum of biologic activities of each of these interactive cytokines.

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