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JAC Advance Access published online on January 13, 2005
Journal of Antimicrobial Chemotherapy, doi:10.1093/jac/dkh526
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JAC © The British Society for Antimicrobial Chemotherapy 2005; all rights reserved
Received September 2, 2004
Revised October 29, 2004
Accepted November 7, 2004

Original article

Whole blood-mediated endothelial permeability and adhesion molecule expression: a model study into the effects of bacteria and antibiotics

Arjan Nooteboom 1*, Cees J. van der Linden 2, and Thijs Hendriks 1

1 Department of Surgery, University Medical Centre Nijmegen, PO box 9101, 6500 HB Nijmegen, The Netherlands
2 Department of Surgery, Atrium Medical Centre, Heerlen, The Netherlands

* To whom correspondence should be addressed.
Arjan Nooteboom, E-mail: a.nooteboom{at}chir.umcn.nl


  Abstract

Aim: To investigate whether the inflammatory response of cultured endothelial cells, as induced by conditioned plasma, depends on the bacterial species or type of antibiotic used for incubation with whole blood.

Materials and methods: Blood from healthy volunteers was stimulated ex vivo with different microorganisms, and with bacteria killed with different antibiotics. The resultant plasmas were incubated on monolayers of cultured human endothelial cells, followed by measurement of their permeability to albumin and expression of E-selectin and intercellular adhesion molecule-1.

Results: Incubation of Escherichia coli in blood yielded plasmas that induced a marked increase in endothelial permeability and E-selectin expression. The response to Bacteroides fragilis or Enterococcus faecalis was generally weaker. Similar effects were observed after incubation of whole blood with lipopolysaccharide (LPS). Much of the permeability and adhesion molecule response to E. coli remained after removal of intact microorganisms from the culture. Whereas antibiotic treatment of E. coli with imipenem or cefuroxime resulted in a divergent production of tumour necrosis factor-{alpha} (TNF-{alpha}) in blood, no significant differences between these treatments were observed with respect to the plasma-induced endothelial response.

Conclusion: Bacteria differ in their capacity to generate a whole blood-mediated increase of endothelial permeability and adhesion molecule expression; this response depends, at least in part, on the presence of soluble bacterial components, such as LPS. Whereas treatment with various antibiotics may generate varying amounts of TNF-{alpha}, these differences are not translated into differences in endothelial permeability or adhesion molecule expression.

Keywords: lipopolysaccharide; cytokines; sepsis; {beta}-lactams; HUVEC.
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