Infect. Immun., Jun 1997, 2272-2277, Vol 65, No. 6
Copyright © 1997, American Society for Microbiology

Saturable CD14-dependent binding of fluorescein-labeled lipopolysaccharide to human monocytes

A Troelstra, P Antal-Szalmas, LA de Graaf-Miltenburg, AJ Weersink, J Verhoef, KP Van Kessel and JA Van Strijp
Eijkman-Winkler Institute for Medical Microbiology, Utrecht University, The Netherlands. a.troelstra@lab.azu.nl

We used rough lipopolysaccharide (ReLPS) to construct a fluorescein- labeled LPS (FITC-LPS) with a very high labeling efficiency that bound to isolated human monocytes in a CD14-dependent fashion and that in this respect behaved indistinctively from native LPS. The CD14- dependent binding could be inhibited either by a 1,000-fold excess of unlabeled LPS or by polymyxin B, bactericidal/permeability-increasing protein, cationic protein 18, or soluble CD14. Although this FITC-LPS preparation no longer possessed the ability to prime neutrophils for the production of reactive oxygen species or to stimulate human monocytes to produce tumor necrosis factor, activation of the Limulus amoebocyte lysate cascade was comparable to activation by native LPS. Binding to monocytes was enhanced by human pooled serum (HPS) or LPS- binding protein (LBP) for LPS concentrations up to 100 ng/ml and was completely CD14 dependent. For LPS concentrations exceeding 100 ng/ml, binding was still partially CD14 dependent, but not HPS or LBP dependent. CD14-dependent association of LPS with monocytes was shown to be totally saturable. In conclusion, we found an HPS- or LBP- dependent binding of FITC-LPS to monocytes that was CD14 dependent at up to 100 ng of LPS per ml, and saturation of binding was shown.

This article has been cited by other articles:

• El Khattabi, M., Adams, H., Heezius, E., Hermans, P., Detmers, F., Maassen, B., van der Ley, P., Tommassen, J., Verrips, T., Stam, J. (2006). Llama Single-Chain Antibody That Blocks Lipopolysaccharide Binding and Signaling: Prospects for Therapeutic Applications. CVI 13: 1079-1086 [Abstract] [Full Text]  
• Berbee, J. F. P., van der Hoogt, C. C., Kleemann, R., Schippers, E. F., Kitchens, R. L., van Dissel, J. T., Bakker-Woudenberg, I. A. J. M., Havekes, L. M., Rensen, P. C. N. (2006). Apolipoprotein CI stimulates the response to lipopolysaccharide and reduces mortality in Gram-negative sepsis. FASEB J. 20: 2162-2164 [Abstract] [Full Text]  
• Fernandez, G. C., Ramos, M. V., Gomez, S. A., Dran, G. I., Exeni, R., Alduncin, M., Grimoldi, I., Vallejo, G., Elias-Costa, C., Isturiz, M. A., Palermo, M. S. (2005). Differential expression of function-related antigens on blood monocytes in children with hemolytic uremic syndrome. J. Leukoc. Biol. 78: 853-861 [Abstract] [Full Text]  
• Arana, M. d. J., Vallespi, M. G., Chinea, G., Vallespi, G. V., Rodriguez-Alonso, I., Garay, H. E., Buurman, W. A., Reyes, O. (2003). Inhibition of LPS-responses by synthetic peptides derived from LBP associates with the ability of the peptides to block LBP-LPS interaction. Innate Immunity 9: 281-291 [Abstract]  
• Triantafilou, M., Triantafilou, K. (2003). Receptor cluster formation during activation by bacterial products. Innate Immunity 9: 331-335 [Abstract]  
• Levels, J. H. M., Abraham, P. R., van den Ende, A., van Deventer, S. J. H. (2001). Distribution and Kinetics of Lipoprotein-Bound Endotoxin. Infect. Immun. 69: 2821-2828 [Abstract] [Full Text]  
• Tanamoto, K.-i., Iida, T., Haishima, Y., Azumi, S. (2001). Endotoxic properties of lipid A from Comamonas testosteroni. Microbiology 147: 1087-1094 [Abstract] [Full Text]  
• Triantafilou, K., Triantafilou, M., Ladha, S., Mackie, A., Dedrick, R. L., Fernandez, N., Cherry, R. (2001). Fluorescence recovery after photobleaching reveals that LPS rapidly transfers from CD14 to hsp70 and hsp90 on the cell membrane. J. Cell Sci. 114: 2535-2545 [Abstract] [Full Text]  
• de Haas, C. J. C., Poppelier, M. J. J. G., van Kessel, K. P. M., van Strijp, J. A. G. (2000). Serum Amyloid P Component Prevents High-Density Lipoprotein-Mediated Neutralization of Lipopolysaccharide. Infect. Immun. 68: 4954-4960 [Abstract] [Full Text]  
• de Haas, C. J. C., van der Zee, R., Benaissa-Trouw, B., van Kessel, K. P. M., Verhoef, J., van Strijp, J. A. G. (1999). Lipopolysaccharide (LPS)-Binding Synthetic Peptides Derived from Serum Amyloid P Component Neutralize LPS. Infect. Immun. 67: 2790-2796 [Abstract] [Full Text]  
• Troelstra, A., de Graaf-Miltenburg, L. A. M., van Bommel, T., Verhoef, J., Van Kessel, K. P. M., Van Strijp, J. A. G. (1999). Lipopolysaccharide-Coated Erythrocytes Activate Human Neutrophils Via CD14 While Subsequent Binding Is Through CD11b/CD18. J. Immunol. 162: 4220-4225 [Abstract] [Full Text]  
• de Haas, C. J. C., van der Tol, M. E., Van Kessel, K. P. M., Verhoef, J., Van Strijp, J. A. G. (1998). A Synthetic Lipopolysaccharide-Binding Peptide Based on Amino Acids 27-39 of Serum Amyloid P Component Inhibits Lipopolysaccharide-Induced Responses in Human Blood. J. Immunol. 161: 3607-3615 [Abstract] [Full Text]  
• Nockher, W. A., Scherberich, J. E. (1998). Expanded CD14+ CD16+ Monocyte Subpopulation in Patients with Acute and Chronic Infections Undergoing Hemodialysis. Infect. Immun. 66: 2782-2790 [Abstract] [Full Text]