Endotoxin-binding substances from human leukocytes and platelets.
We have found whole human platelets, granulocytes, and mononuclear leukocytes to possess high affinity for the toxic lipopolysaccharide from all gram-negative bacteria tested. We have extracted these cells and platelets with n-butanol-water; all endotoxin-binding activity resided in the organic phase. These endotoxin-binding extracts did not block serologically active groupings on  endotoxins or receptors on the erythrocytes. The specificity of these still crude materials was less that that of the highly purified erythrocyte lipopolysaccharide receptor previously described by us, since they bound some bacterial antigens not related to endotoxins. Depending on source, the n-butanol extracts contained 40 to 52% glycerophosphatides (most active), 15 to 22% sphingomyelin, 17% cholesterol, less than 2 to 5% triglycerides, and 7 to 13% inactive peptide. The most active substances in the n-butanol extract were soluble in petroleum ether, whereas the peptide and sphingomyelin were not. Thus, no constituent protein, carbohydrate, or nucleic acid was present in the most highly active material. Polyacrylamide gel electrophoresis of the petroleum ether-soluble material showed for each extract one lipid band only, which was well defined and migrated similarly to phosphatidyllipids. Because of the lipidic nature of the inhibitory substances from leukocytes and platelets we also tested the lipid A component of bacterial endotoxins and some of its derivatives. Lipid A inhibited endotoxin coating of erythrocytes. De-O-acylation of lipid A left amide-linked 3-D-hydroxymyristic acid intact and increased the inhibitory activity of lipid A 20-fold. Complete de-O- and de-N-acylation destroyed its inhibitory effect.
D000942:Antigens, Bacterial / Q000302:isolation & purification; D001665:Binding Sites; D001667:Binding, Competitive; D001792:Blood Platelets / Q000032:analysis*; D002469:Cell Separation; D002784:Cholesterol / Q000032:analysis; D004731:Endotoxins / Q000032:analysis*; D004912:Erythrocytes / Q000276:immunology; D005227:Fatty Acids / Q000032:analysis; D006098:Granulocytes; D006385:Hemagglutination Inhibition Tests; D006801:Humans; D007962:Leukocytes / Q000032:analysis*; D008070:Lipopolysaccharides / Q000494:pharmacology; D008214:Lymphocytes; D009000:Monocytes; D010455:Peptides / Q000032:analysis; D011955:Receptors, Drug; D013109:Sphingomyelins / Q000032:analysis; D014280:Triglycerides / Q000032:analysis
D000942:Antigens, Bacterial; D004731:Endotoxins; D005227:Fatty Acids; D008070:Lipopolysaccharides; D010455:Peptides; D011955:Receptors, Drug; D013109:Sphingomyelins; D014280:Triglycerides; D002784:Cholesterol