Monoclonal IgM antiphosphatidylserine antibody reacts against cytoskeleton-like structures in cultured human umbilical cord endothelial cells.
OBJECTIVE
It has been proposed that antibodies against phospholipid-dependent antigens (aPLs), induce recurrent pregnancy loss and thrombosis through modulation of endothelial cell function, yet aPLs have not been conclusively shown to bind with endothelial cells.


METHODS
Using indirect immunofluorescence we investigated the anti-endothelial cell reactivity of three monoclonal antibodies that differentiate between the phospholipids cardiolipin (CL) and phosphatidylserine (PS): BA3B5C4 (CL+/PS+); 3SB9b (CL-/PS+); and D11A4 (CL+/PS-). Cultured umbilical cord endothelial cells were prepared without fixation or with cold acetone fixation.


RESULTS
None of the aPLs reacted with endothelial cells prepared without fixation. 3SB9B reacted strongly with cytoskeletal-like components in acetone-fixed cells, whereas BA3B5C4 and D11A4 were unreactive. The cytoskeletal-like binding of 3SB9b was completely blocked by a monoclonal antibody against vimentin, whereas antibodies against tubulin or actin were not inhibitory. Lipid extraction of the cells destroyed the 3SB9b reactive antigen without affecting the reactivity of anti-vimentin.


CONCLUSIONS
These results suggest that phospholipid-dependent antigenic determinants are not expressed on the surface of resting endothelial cells but that a PS-dependent antigenic determinant is associated with endothelial cell intermediate filaments.
Lin
L
L
Department of Microbiology and Immunology, Wright State University School of Medicine, Dayton, Ohio 45435, USA.
Shroyer
L
L
Walter
A
A
Lyden
T W
TW
Ng
A K
AK
Rote
N S
NS
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000918:Antibody Specificity; D016736:Antiphospholipid Syndrome / Q000276:immunology*; D002308:Cardiolipins / Q000276:immunology; D002460:Cell Line; D002478:Cells, Cultured; D003598:Cytoskeletal Proteins / Q000276:immunology; D003599:Cytoskeleton / Q000276:immunology*; D004730:Endothelium, Vascular / Q000276:immunology* / Q000648:ultrastructure; D000939:Epitopes / Q000276:immunology; D006801:Humans; D007075:Immunoglobulin M / Q000276:immunology*; D007614:Macropodidae; D008856:Microscopy, Fluorescence; D010718:Phosphatidylserines / Q000276:immunology*; D014470:Umbilical Cord; D014746:Vimentin / Q000145:classification / Q000276:immunology*
D000911:Antibodies, Monoclonal; D002308:Cardiolipins; D003598:Cytoskeletal Proteins; D000939:Epitopes; D007075:Immunoglobulin M; D010718:Phosphatidylserines; D014746:Vimentin