Differential binding of bacterial lipopolysaccharide to bovine peripheral-blood leukocytes

Abstract

The response of mammalian monocytes and macrophages to bacterial lipopolysaccharide (LPS) may be influenced by serum factors that increase or decrease the propensity of LPS to bind to cell surfaces. We used fluorescence flow cytometric analysis to investigate the capability of bovine peripheral-blood leukocytes to bind LPS in the presence or absence of bovine serum. At all concentrations of FITC-LPS tested (LPS from E. coli 0111:B4; 10 ng/ml, 100 ng/ml, 1 micrograms/ml), monocytes, lymphocytes, and granulocytes bound more FITC-LPS in the presence of 10% bovine serum than in serum-free conditions (P < 0.01). At the intermediate concentration tested (100 ng/ml), monocytes displayed a relative fluorescence intensity (RFI) of 2.27 +/- 1.24 units without serum and 17.48 +/- 8.05 with 10% serum. Values for granulocytes were similar to those of monocytes, 3.55 +/- 1.31 without and 19.24 +/- 6.93 with serum, and values for lymphocytes were 1.89 +/- 0.47 RFI units without serum and 6.27 +/- 2.61 RFI units with serum. At 10 ng/ml and 1 microgram/ml FITC-LPS the RFI of monocytes and granulocytes were also similar and not significantly different, and both bound significantly more LPS than lymphocytes (P < 0.01). When 100 ng/ml FITC-LPS was coincubated with leukocytes, 10% serum, and a 100-fold excess of unlabeled LPS, the amount of FITC-LPS bound to monocytes was reduced from 23.99 to 7.23 RFI units (P < 0.01), reduced from 22.00 to 7.30 RFI units with granulocytes (P < 0.05), and reduced from 7.51 to 2.29 RFI units (P < 0.10) with lymphocytes. These data demonstrate that factors in bovine serum significantly amplify the association of LPS with peripheral-blood leukocytes and that increased binding of LPS is greatest with monocytes and granulocytes.