N-glycosidase treatment of Colo 205 cells interferes with hIFN-gamma induced HLA-DR expression.

Abstract

We investigated the effects of N-glycosidase treatment on the interferon-gamma (IFN-gamma) induced HLA-DR expression of Colo 205 cells. N-glycosidase treatment resulted in a significant decrease of IFN-gamma induced HLA-DR specific immunofluorescence intensity ranging from complete reduction to approximately 30% of that of untreated control cells depending on the IFN-gamma dose. IFN-gamma binding studies showed that this was due to a severe reduction in IFN-gamma binding capacity of N-glycosidase treated cells. Since the number of cell membrane IFN-gamma receptors (IFN-gamma-Rs) was virtually unchanged as revealed by immunofluorescence analysis with a hIFN-gamma-R specific monoclonal antibody this indicates that N-linked carbohydrates play an important role in signal transduction and ligand binding capacity of the hIFN-gamma-R and strengthens the view that carbohydrate moieties of receptor proteins are of greater functional significance than originally anticipated.