Metabolic carbohydrate labelling of glycoproteins from mitogen-stimulated mouse lymphocytes. Glycoproteins as biochemical markers for lymphocyte subpopulations
van Eijk, R.V.; Rosenfelder, G.; Mühlradt, P.F.
European Journal of Biochemistry 101(1): 185-193
ISSN/ISBN: 0014-2956 PMID: 315872 Accession: 005880255
CBA/J mouse lymphocytes from thymus, spleen and lymph nodes were separated into fractions enriched in T (thymus-derived) or B (bone marrow-derived) cells. The cells were stimulated with T cell mitogens (concanavalin [con] A or phytohemagglutinin [PHA]), B cell mitogens (lipopolysaccharide [LPS] or lipoprotein) and pokeweed mitogen to incorporate tritiated mannose, galactose or fucose. Mitogen-induced incorporation of carbohydrates into acid-precipitable material was specific for the cell population. Carbohydrate-labeled cells were extracted with dodecyl sulfate buffer and the extracts subjected to polyacrylamide gel electrophoresis and subsequent autoradiography. Depending on the carbohydrate used for labeling, several glycoproteins specific for activated T cells (GP 200, GP 170, GP 125) and activated B cells (GP 210, GP 85 and GP 75) were noted. T or B cell subpopulations could be distinguished by their glycoprotein patterns. Con-A-reactive T cells from thymus, spleen and lymph nodes comprise different subpopulations. Con A and PHA stimulate the same T cell subpopulations in spleen and lymph nodes. LPS and lipoprotein stimulate different B cell subpopulations of the spleen. LPS stimulated or lipoprotein-stimulated B cells from lymph nodes differ from those in the spleen in that the latter show a markedly higher rate of labeling for GP 75, suggesting a higher rate of immunoglobulin(Ig)M synthesis.