Biosynthesis of glycosphingolipids by human myeloid leukemia cells

Abstract

We have performed comparative studies of the neutral glycosphingolipids synthesized by three human myeloid leukemia cell lines, K562, KG1, and HL-60, which were metabolically labeled with [14C]galactose, to evaluate changes in neutral glycosphingolipid synthesis with myeloid cell differentiation. Individual neutral glycosphingolipids containing one to four sugars were purified by a combination of the following methods: diethylaminoethyl-Sephadex column chromatography, acetylation-Florisil column chromatography, and high-performance liquid chromatography using an Iatrobead column. Compounds with one sugar were analyzed by thin-layer chromatography on borate plates. This analysis showed that HL-60 cells synthesize only glucosylceramide, whereas K562 and KG1 cells synthesize predominately glucosylceramide, but also a small amount of galactosylceramide. Compounds with two to four sugars were characterized by treatment with exo- and endoglycosidases. The results showed that K562 and KG1 cells are similar to cells from patients with acute leukemia in expressing two series (globo and neolacto) of natural glycosphingolipids, whereas the HL-60 cells are similar to mature human myeloid cells in expressing only one series (neolacto). Therefore, human myeloid leukemia cells blocked at different stages of differentiation vary in their ability to synthesize neutral glycosphingolipids.

Association of Shiga toxin glycosphingolipid receptors with membrane microdomains of toxin-sensitive lymphoid and myeloid cells

“…From these early studies and following investigations using leukocyte-derived cell lines, which may be regarded as cells frozen at different stages of lymphoid and myeloid development, it became apparent that the human myeloid cell lines THP-1 and HL-60 profoundly differ in their neutral GSL repertoire. The monocyte-like THP-1 cell line produced globo-series neutral GSLs (beside others), whereas the granulocyte-like HL-60 cell line synthesized only neo lacto-series GSLs, indicating that GSLs of the globoseries are associated with the monocytic lineage (90)(91)(92)(93)(94). The lymphoid Raji and Jurkat cell lines showed a similar difference regarding globo-series GSLs.…”

“…Neutral GSLs have proved in the past to be useful cell surface markers for characterizing distinct human leukocyte subpopulations such as human blood lymphocytes, monocytes, and neutrophilic granulocytes ( 88,91 ). From these early studies and following investigations using leukocyte-derived cell lines, which may be regarded as cells frozen at different stages of lymphoid and myeloid development, it became apparent that the human myeloid cell lines THP-1 and HL-60 profoundly differ in their neutral GSL repertoire.…”

Glycosphingolipid expression in acute nonlymphocytic leukemia: common expression of shiga toxin and parvovirus B19 receptors on early myeloblasts

“…Unlike earlier studies, 27,29-31 we were able to examine GSL expression relative to myeloid differentiation, as defined by histologic and immunophenotypic markers. Based on our data and those of earlier investigators, 27,29 we present a modified model of neutral GSL differentiation during human myelopoiesis and discuss the implications for mature granulocyte function, parvovirus B19 infection, and transplantation.…”

“…The gala-series GSL, galactosylceramide (GalCer), is reportedly expressed by the myeloid leukemia cell lines K562 and KG1 27,70 and is a receptor for HIV. 17 A second gala-series GSL, galabiosylceramide (Gal 2 Cer), is a receptor for Stx's and has been reported to be a marker of mature neutrophils.…”

“…26 Gb 3 and/or Gb 4 has also been identified in 3 immortalized human myeloid leukemia cell lines, KG1, K526, and THP-1, as well as a small number of clinical acute nonlymphocytic leukemia (ANLL) samples. [27][28][29][30][31][32] In addition, there is evidence that parvovirus B19, which recognizes Gb 4 , can infect early myeloid cells, decrease myeloid progenitors, and inhibit granulocyte-monocyte colony (CFU-GM) formation in vivo. 18,[33][34][35][36] In contrast, globo GSLs are not expressed by either mature neutrophils or chronic myeloid leukemia (CML) cells.…”