The intermediate filament protein vimentin is a potential target antigen for autoantibodies in some infectious and autoimmune diseases. Because endothelial cells contain an extensive interconnecting cytoplasmic network of vimentin, we examined the relationship between anti-vimentin and anti-endothelial cell antibodies (AECAs). We measured the level of anti-vimentin antibodies in patients with systemic autoimmune diseases (n=42), healthy blood donors (n=58), and patients with acute inflammation and showing anti-vimentin immunofluorescence (n=50). AECAs were detected by indirect immunofluorescence in human umbilical vein endothelial cells. Anti-vimentin antibodies were detected by indirect immunofluorescence in HEp-2 cells, and levels of anti-vimentin antibodies were measured using an enzyme-linked immunosorbent assay. We did not find significant differences in anti-vimentin levels between AECA-positive and-negative samples for any of the groups; however, serum anti-vimentin antibody levels were significantly higher in patients with acute non-autoimmune inflammatory diseases than in patients with systemic autoimmune diseases or healthy controls. There were no differences between the levels of anti-vimentin antibodies between patients with systemic autoimmune diseases and healthy individuals. The anti-vimentin antibodies levels also did not correlate with the AECA positivity.