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. 1977 Jan;15(1):156–164. doi: 10.1128/iai.15.1.156-164.1977

Binding of bacterial endotoxin to murine spleen lymphocytes.

S Kabir, D L Rosenstreich
PMCID: PMC421342  PMID: 299851

Abstract

The early events in lipopolysaccharide (LPS)-induced B-cell activation were investigated by studying the binding of 14C-labeled LPS to murine lymphocytes in vitro. In these studies we utilized intrinsically labeled 14C-labeled LPS from Salmonella minnesota or the 14C-labeled glycolipid derived from the Re mutant of S. minnesota (R595). Bone marrow-derived (B) lymphocytes bound more LPS than did thymus-derived (T) lymphocytes. Binding of LPS to murine spleen lymphocytes from strain C3H/HeN was compared with the binding to spleen lymphocytes from strain C3H/HeJ, a strain resistant to certain biological activities of LPS including mitogenesis. Spleen cells from both strains bound LPS equally well, suggesting that unresponsiveness of C3H/HeJ mice to LPS is due to factors other than a defect in binding of LPS. LPS binding to cells appeared to be due to a nonspecific interaction between the lipid moiety of LPS and the lipid components of the cell membrane. Thus, the highly lipophilic, polysaccharide-deficient glycolipid from R595 bound at least 20 times better than did LPS. Furthermore, partial removal of cell surface proteins with trypsin or sialic acids with neuraminidase enhanced glycolipid binding, suggesting that binding is not through a protein- or sialic acid-containing receptor. The binding of glycolipid to lymphocytes was only partially specific since unlabeled glycolipid R595, lipid A, and LPS did not completely inhibit the uptake of 14C-labeled glycolipid R595. In addition, binding could be inhibited by a nonmitogenic phospholipid (phosphatidyl ethanolamine), which also is consistent with a nonspecific lipid-lipid interaction. Experiments were performed to determine the relationship of LPS binding to lymphocyte activation in the lymphocytes. The process of activation of lymphocytes by LPS was a slow one, since LPS was required to be present in culture for at least 24 h in order to obtain significant lymphocyte activation, suggesting that the amounts of LPS bound earlier are either quantitatively or qualitatively insufficient to irreversibly activate the cell.

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Selected References

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