Alpha 1,4galactosyltransferase activity and Gb3Cer expression in human leukaemia/lymphoma cell lines

We have used two methods to evaluate the level of expression of Gb3Cer in several human leukaemia/lymphoma cell lines representative of the myeloid (K562, KG-1, HL-60, and lymphoid (Reh, Daudi, Raji, RPMI 8226, CCRF-CEM, MOLT-4) lineages blocked at varied stages of differentiation. TLC immunostaining of glycolipid extracts with a monoclonal antibody, 12-101, and FACS analysis with the same antibody were used to demonstrate that the expression of Gb3Cer in neoplastic myeloid and lymphoid cells is both lineage and differentiation dependent. As a possible control point in the regulated expression of Gb3Cer we have investigated the first committed step in the synthesis of globo series glycosphingolipids that involves UDP-Gal:LacCer alpha (1,4)-galactosyltransferase (alpha 1,4GalT). We present the first characterization of this enzyme in a human myeloid cell line using an ELISA-based assay, which was subsequently used to measure alpha 1,4GalT activity in the human leukaemia/lymphoma cell lines. In general, there is a positive correlation between the levels of endogenous Gb3Cer and the level of the alpha 1,4 GalT activity. However, in two cases (KG-1 and CCRF-CEM) the level of enzyme activity did not correspond to the level of Gb3Cer expression.