Murine
interleukin-3 (IL-3)-associated expression of
gangliosides has been investigated using a gene transfection technique. A murine
IL-3 cDNA was introduced into the parental NFS60-17 cells that was exclusively dependent on
IL-3. We analyzed the
glycosphingolipids from the parental cells and the transfected cells by fast atom bombardment mass spectrometry analyses and/or immunostaining techniques using specific
antibodies. Two major
gangliosides, IV3NeuAc-GgOse4Cer (
GM1b) and IV3-NeuAc,III6NeuAc-GgOse4Cer (GD1 alpha), were expressed, in the parental cells. By contrast, in the
IL-3 gene-transfected cells, a
ganglioside IV3NeuAc,II3NeuAc-GgOse4Cer (GD1a) was strikingly expressed, in addition to
GM1b and GD1 alpha that were already present in the parental cells. In spite of various IL-3-secreting capabilities, all transfectants investigated have exhibited the same
ganglioside patterns and expressed GD1a. Furthermore, the appearance of GD1a was a consequence of the up-regulation of a single
glycosyltransferase,
CMP-NeuAc:
lactosylceramide alpha 2-->3-sialytransferase (
GM3 synthase). Activities of the other downstream
glycosyltransferases that were involved in GD1a synthesis were not significantly different between the parental and the transfected cells. According to these data, the progression of
tumor stage by the acquisition of autonomous cell growth ability after
IL-3 gene transfection resulted in dramatic changes in cell surface
gangliosides and their biosynthetic pathways. GD1a could be considered as an IL-3-associated
ganglioside and was expressed in a tight connection with a single
glycosyltransferase (
GM3 synthase) up-regulation and with
IL-3 expression in murine
myelogenous leukemia cells.