The effects of monensin on blood-borne arrest and glycosylation of BL/VL3 lymphoma cells.
We have previously shown that inhibitors of N-glycan processing alter both the cell surface carbohydrates and the homing properties in lymphoid cells. We have now studied the effects of the ionophore monensin (MON) on these parameters. Arrest in the spleen of [111In]-labelled BL/VL3 murine T lymphoma cells, injected intravenously was clearly reduced if the cells had been cultured for 24 h in the presence of monensin (0.1-1.0 microgram ml-1). We have characterized glycopeptides from BL/VL3 murine T lymphoma cells. Following labelling with tritiated precursors (fucose, mannose, galactose, glucosamine), surface glycopeptides from BL/VL3 murine T lymphoma cells, were released by trypsin and separated by gel filtration on Bio-Gel P6 and by affinity chromatography on immobilized lectins. After treatment with MON, a class of high molecular mass glycopeptides was no longer found. There were less complex and more high mannose glycans, as a consequence of a reduction of terminal glycosylation (sialylation, fucosylation or incorporation of N-acetyl-glucosamine). Similar findings were obtained with immunoprecipitated Thy-1 antigen. However, as estimated by flow cytometry analysis, the cell surface expression of Thy-1 was not reduced in MON-treated cells. Taken together our results show that cell surface oligosaccharides are modified dramatically, but that at least, certain cell surface antigens are present in normal amounts. It is tempting to speculate that changes in glycosylation account for the abnormal homing properties of MON-treated cells.
Department of General Chemistry I, Université Libre de Bruxelles, Belgium.
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Di Virgilio
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Rampelberg
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Greimers
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G
Schnek
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Hooghe
D000818:Animals; D000954:Antigens, Surface / Q000096:biosynthesis; D001775:Blood Circulation; D002240:Carbohydrate Sequence; D002462:Cell Membrane / Q000737:chemistry / Q000187:drug effects / Q000378:metabolism*; D002465:Cell Movement / Q000187:drug effects*; D002846:Chromatography, Affinity; D006020:Glycopeptides / Q000187:drug effects / Q000302:isolation & purification / Q000378:metabolism*; D006031:Glycosylation / Q000187:drug effects; D016399:Lymphoma, T-Cell / Q000378:metabolism*; D008562:Membrane Glycoproteins / Q000096:biosynthesis; D051379:Mice; D008969:Molecular Sequence Data; D008985:Monensin / Q000494:pharmacology*; D011134:Polysaccharides / Q000302:isolation & purification; D018800:Thy-1 Antigens; D014407:Tumor Cells, Cultured
D000954:Antigens, Surface; D006020:Glycopeptides; D008562:Membrane Glycoproteins; D011134:Polysaccharides; D018800:Thy-1 Antigens; D008985:Monensin