Neutral and sialosyl glycosphingolipid composition and metabolism of human T-lymphoblastic cell line MOLT-3 cells: distinctive changes as markers specific for their differentiation.

Changes in the composition and metabolism of glycosphingolipid (GSL), which is one of the cell surface constituents, during cell differentiation of human T-lymphoblastic leukemia cell line MOLT-3 cells were examined with special reference to their alterations in E rosette-forming capacity and expression of surface antigens specific for T-cell lineage. Three molecular species of neutral GSL and greater than or equal to 13 molecular species of acidic sialosyl-GSL (ganglioside) were detectable on high-performance thin-layer chromatography (HPTLC) in untreated MOLT-3 cells. The major components were ceramide monohexoside and gangliosides GM3 and GD1a. When the cells were induced by 12-O-tetradecanoyl phorbol 13-acetate (TPA) to differentiate into more mature T cells, the ganglioside composition changed distinctively, and the total ganglioside content increased considerably; mono-, di-, and tri-sialosyl gangliosides concomitantly showed significant increase, but no new molecular species of GSL specific for the differentiation were detected. The activity of one sialyltransferases, CMP-sialic acid:CDH sialyltransferase, which synthesizes ganglioside GM3 and the total sialic acid content of the cell surface, parallelled the extent of cell differentiation. Examination of another human T-lymphoblastic leukemia cell line, HPB-ALL, indicated that TPA could also induce the cells to differentiate along T-cell lineage and that changes in the ganglioside pattern during differentiation are similar to those of MOLT-3 cells. The results indicate that human T-lymphoid cell differentiation intimately involves elongation of neutral oligosaccharide-moieties and the addition of sialic acid residues to gangliosides, resulting in more mature T cells containing higher gangliosides. Both the sialyltransferase activity and the sialic acid content, as well as the ganglioside pattern, might be new biochemical markers specific for human T-lymphoblastic cell differentiation.