Heparin-binding protein (CAP37) is internalized in monocytes and increases LPS-induced monocyte activation.
Previous studies have shown that the neutrophil-derived heparin-binding protein (HBP), also known as CAP37 or azurocidin, potentiates the LPS-induced release of proinflammatory cytokines (TNF-alpha, IL-1, and IL-6) from isolated human monocytes. To date, the mechanisms by which HBP enhances LPS-induced monocyte activation have not been elucidated, and it is not known whether HBP also increases the LPS-induced production of other bioactive substances. We studied human monocytes activated by recombinant human HBP and LPS and their interaction with the LPS receptor CD14. We hypothesized that the stimulatory effect of HBP on the LPS-induced release of proinflammatory mediators from monocytes was mediated by specific binding of HBP to monocytes, which resulted in an up-regulation of CD14. Our results demonstrated that HBP alone (10 microg/ml) stimulated the production of TNF-alpha from isolated monocytes. In addition, HBP had an additive effect on LPS-induced production of TNF-alpha and PGE2, suggesting a generalized monocyte activation. We used flow cytometry to demonstrate that HBP had a high affinity to monocytes but not to the LPS receptor CD14, and experiments performed at 4 degrees C indicated an energy-dependent step in this process. Confocal microscopy showed that monocytes internalize HBP within 30 min. These data suggest that mechanisms other than increased CD14 expression are responsible for the enhanced release of TNF-alpha or PGE2 in response to HBP and LPS.
Heinzelmann
M
M
Department of Physiology and Biophysics, The Price Institute of Surgical Research, University of Louisville, School of Medicine, KY 40292, USA. mheinzelmann@bluewin.ch
Mercer-Jones
M A
MA
Flodgaard
H
H
Miller
F N
FN
D000276:Adjuvants, Immunologic / Q000502:physiology*; D023181:Antimicrobial Cationic Peptides; D001798:Blood Proteins / Q000378:metabolism* / Q000502:physiology; D018821:CD18 Antigens / Q000096:biosynthesis / Q000502:physiology; D002352:Carrier Proteins / Q000378:metabolism* / Q000502:physiology; D006493:Heparin / Q000378:metabolism* / Q000494:pharmacology; D006801:Humans; D007962:Leukocytes / Q000378:metabolism; D018950:Lipopolysaccharide Receptors / Q000096:biosynthesis / Q000502:physiology; D008070:Lipopolysaccharides / Q000494:pharmacology*; D008262:Macrophage Activation / Q000187:drug effects*; D009000:Monocytes / Q000378:metabolism*; D011485:Protein Binding / Q000276:immunology; D013696:Temperature
C062991:AZU1 protein, human; D000276:Adjuvants, Immunologic; D023181:Antimicrobial Cationic Peptides; D001798:Blood Proteins; D018821:CD18 Antigens; D002352:Carrier Proteins; D018950:Lipopolysaccharide Receptors; D008070:Lipopolysaccharides; D006493:Heparin