A simple method for the assessment of macrophage scavenger receptor-ligand interaction: adherence of erythrocytes coated with oxidized low density lipoprotein and modified albumin to macrophages.
A simple method is described for the assessment of the binding of macrophage scavenger receptor and its ligands such as oxidized low density lipoprotein (ox-LDL) and maleylated bovine serum albumin (m-BSA). In this method, the binding of ox-LDL or m-BSA to macrophages is observed as the adherence of erythrocytes precoated with ox-LDL or m-BSA under phase-contrast microscopy. Erythrocytes were coated with native LDL or ox-LDL simply by incubating mouse erythrocytes with LDL. The ox-LDL-coated erythrocytes attached to the monolayer of mouse peritoneal macrophages without phagocytosis at 37 degrees C, while native LDL-coated erythrocytes did not. The extent of the adherence of the ox-LDL-coated erythrocytes to the macrophages was conveniently expressed as the proportion of the macrophages binding the coated erythrocytes. An optimal concentration of ox-LDL for erythrocyte coating giving maximum erythrocyte adherence to macrophages was 10 micrograms/ml as protein at an erythrocyte concentration of 2%. The binding of the ox-LDL-coated erythrocytes to the macrophages could be inhibited by ligands for scavenger receptor, and reflected the extent of LDL oxidation. Mouse erythrocytes were successfully coated with m-BSA using chromium(III) ion as adsorbent. The m-BSA-coated erythrocytes attached to the macrophages, while native BSA-coated erythrocytes did not. The adherence was inhibitable with ligands for scavenger receptor, and reflected the extent of maleylation. The method would be useful particularly for measurement of the ligand-binding activity of the receptor, and the receptor-binding activity of the ligands.
Beppu
M
M
Tokyo College of Pharmacy, Japan.
Hora
M
M
Kikugawa
K
K
D000042:Absorption; D000418:Albumins / Q000737:chemistry; D000818:Animals; D001665:Binding Sites; D002448:Cell Adhesion; D004912:Erythrocytes / Q000378:metabolism*; D005557:Formaldehyde / Q000737:chemistry; D008077:Lipoproteins, LDL / Q000737:chemistry*; D017737:Macrophages, Peritoneal / Q000378:metabolism*; D051379:Mice; D008858:Microscopy, Phase-Contrast; D010084:Oxidation-Reduction; D010587:Phagocytosis; D011869:Radioligand Assay; D011971:Receptors, Immunologic / Q000378:metabolism*; D051116:Receptors, Scavenger; D012710:Serum Albumin, Bovine / Q000737:chemistry*
D000418:Albumins; D008077:Lipoproteins, LDL; D011971:Receptors, Immunologic; D051116:Receptors, Scavenger; C049877:maleylalbumin; D005557:Formaldehyde; D012710:Serum Albumin, Bovine