We have previously demonstrated that amounts of ganglioside GM3 are markedly higher in human atherosclerotic lesions compared to that in non-diseased arterial tissue. Because the fatty acid composition of GM3 in blood plasma low density lipoproteins (LDL) and the fatty acid composition of GM3 in atherosclerotic lesions differed, we hypothesized that, in addition to GM3 originating from LDL infiltrating the arterial wall from the blood, excessive GM3 may be synthesized locally in atherosclerotic lesions. In the present work, using an anti-GM3 antibody developed by us, we showed that the levels of GM3 synthase in membrane fractions isolated from the atherosclerotic intima were higher compared to those in non-diseased arterial tissue. Using an immunohistochemical approach, we examined the expression of GM3 synthase in sections of atherosclerotic plaques and non-diseased arterial wall. GM3 synthase immunopositivity was found to be low in non-diseased arterial intima but large numbers of GM3 synthase-immunopositive cells were observed in atherosclerotic plaques. GM3 synthase was overexpressed by macrophages and dendritic cells and double immunostaining demonstrated cellular co-localization of GM3 synthase and GM3. Further in vitro experiments showed that both monocyte-derived dendritic cells and macrophages expressed high levels of GM3 synthase. The findings of the present study indicate that, at least partially, excessive amounts of GM3 in atherosclerotic lesions can be synthesized by macrophages and dendritic cells directly within the arterial wall.