Lipoprotein lipase in heart cell cultures is suppressed by bacterial lipopolysaccharide: an effect mediated by production of tumor necrosis factor.
Exposure of rat heart cell cultures, consisting mainly of nonbeating mesenchymal cells, to 50 ng/ml of bacterial lipopolysaccharide (LPS) for 24 h resulted in a more than 80% reduction in lipoprotein lipase activity. The loss of enzymic activity was accompanied by a concomitant reduction in enzyme protein, as shown by immunoblotting. Addition of LPS to the culture medium resulted also in the production of tumor necrosis factor (TNF), and the fall in lipoprotein lipase in LPS-treated cultures could be prevented by an antibody to TNF. Addition of recombinant human TNF to the heart cell cultures also depressed lipoprotein lipase activity. LPS treatment of preadipocytes in culture resulted in a fall in lipoprotein lipase activity and TNF production. Since TNF is known as a macrophage product, the cultures were tested for phagocytic capacity, and only 0.2-1.3% of the cells were shown to engulf Staphylococcus albus. Immunofluorescent staining with monoclonal antibodies OX-1, which identify leukocyte common antigen, was negative, and only 0.1 +/- 0.07% of the cells were positive after staining with OX-42 antibody to iC3b receptor. Both antibodies stained more than 98% of rat peritoneal macrophages used as controls. Since LPS treatment of macrophages at numbers comparable to or exceeding the number of phagocytic cells present in the heart cell cultures did not induce measurable amounts of TNF, it is suggested that in the heart cell cultures, TNF may be produced by cells other than macrophages.
Friedman
G
G
Department of Medicine B, Hadassah University Hospital, Jerusalem, Israel.
Gallily
R
R
Chajek-Shaul
T
T
Stein
O
O
Shiloni
E
E
Etienne
J
J
Stein
Y
Y
D000818:Animals; D000911:Antibodies, Monoclonal; D002478:Cells, Cultured; D005455:Fluorescent Antibody Technique; D007930:Leucine / Q000378:metabolism; D008070:Lipopolysaccharides / Q000494:pharmacology*; D008071:Lipoprotein Lipase / Q000378:metabolism*; D009206:Myocardium / Q000201:enzymology*; D010587:Phagocytosis; D051381:Rats; D013210:Staphylococcus; D014409:Tumor Necrosis Factor-alpha / Q000096:biosynthesis*
D000911:Antibodies, Monoclonal; D008070:Lipopolysaccharides; D014409:Tumor Necrosis Factor-alpha; D008071:Lipoprotein Lipase; D007930:Leucine