Grafting of triggering site onto lymphocytes; distribution of grafted dinitrophenyl groups on cell surface glycoproteins and glycolipids

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Abstract
Quantitative investigation of membrane-bound sialoglycoconjugates on lymphocyte surface was performed by chemical modification of the sialic acid residues with radioactive N4-dinitrophenyl-l-2,4-diaminobutyric acid hydrazide (DNP-DABH). This labeled both glycoproteins and glycolipids with concomitant preservation of the mitogenic activity by multivalent hapten binding protein (anti-DNP antibody). Under conditions where maximum stimulation of thymocytes occurred radioactive DNP-DABH labeled 1.1 × 107 glycolipids molecules/cell but, only 3 × 106 glycoproteins molecules/cell. When B lymphocytes, which do not undergo DNP-mediated stimulation were used, glycolipids labeling could not be detected. Major differences between stimulation committed and non-committed DNP-modified lymphocytes was the amount of ligand attached to the cell surface sialoglycolipids (gangliosides).