This Article Full Text (PDF) References Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Alert me to new issues of the journal Add to Saved Citations Download to citation manager Request Permissions Request Reprints Add to My Marked Citations Citing Articles Citing Articles via Google Scholar Citing Articles via Scopus Google Scholar Articles by Haslberger, A.G. Articles by Lubitz, W. Search for Related Content PubMed Articles by Haslberger, A.G. Articles by Lubitz, W. Social Bookmarking                         What's this?

Bacterial cell envelopes (ghosts) and LPS but not bacterial S-layers induce synthesis of immune-mediators in mouse macrophages involving CD14

Institute of Microbiology and Genetics, Biocenter, University of Vienna, Vienna, Austria

H.J. Mader

Institute of Microbiology and Genetics, Biocenter, University of Vienna, Vienna, Austria

M. Schmalnauer

Institute of Microbiology and Genetics, Biocenter, University of Vienna, Vienna, Austria

G. Kohl

Institute of Microbiology and Genetics, Biocenter, University of Vienna, Vienna, Austria

M.P. Szostak

Institute of Microbiology and Genetics, Biocenter, University of Vienna, Vienna, Austria

P. Messner

Zentrum für Ultrastrukturforschung und Ludwig Boltzmann-Institut für Molekulare Nanotechnologie, Universität für Bodenkultur, Vienna, Austria

U.B. Sleytr

Zentrum für Ultrastrukturforschung und Ludwig Boltzmann-Institut für Molekulare Nanotechnologie, Universität für Bodenkultur, Vienna, Austria

G. Wanner

Institute of Botany, LM University of Munich, Munich, Germany

S. Fürst-Ladani

Ludwig-Boltzmann Institut für experimentelle und klinische Traumatologie, Lorenz-Böhler Krankenhaus, Vienna, Austria

W. Lubitz

Institute of Microbiology and Genetics, Biocenter, University of Vienna, Vienna, Austria

The synthesis of inflammatory mediators in human macrophages/monocytes seen after stimulation with lipopolysaccharide (LPS) involves the binding of CD14 to LPS complexed to lipopolysaccharide binding protein (LBP). The binding mechanisms of different LPS domains to LBP and CD14, as well as the interaction of the entire bacterial cell wall and its components with CD14 and LBP, are poorly understood. We, therefore, studied the effects of anti-mouse CD14 antibodies on the synthesis of TNF and PGE₂ in RAW 264.7 mouse macrophages stimulated by bacterial cell envelopes (ghosts) of Escherichia coli 026:B6 and Salmonella typhimurium C5, LPS, lipid A, and crystalline bacterial cell surface layer (S-layer) preparations. Ghosts and S-layers, with distinct activities on the immune-system, are presently under investigation for their use as vaccines. Whereas LPS and E. coli ghosts exhibited a strong endotoxic activity in the Limulus amoebocyte lysate assay, the endotoxic activity of S-layer preparations was several orders of magnitude lower. LPS, ghosts, and bacterial S-layers all induced TNF and PGE₂ synthesis as well as the accumulation of TNF mRNA. Pre-incubation with anti-mouse CD14 antibodies resulted in a dose-dependent inhibition of TNF and PGE ₂ synthesis after stimulation by LPS, lipid A (30-50%) and ghosts (40-70%). The bacterial S-layer-induced mediator synthesis remained unchanged following the addition of anti-mouse CD14 antibodies. Reproducible differences could be observed for the inhibition of TNF induced by LPS of different species by anti-CD14. Adding fetal calf serum (FCS) strongly enhanced the release of cell mediators stimulated by low doses of LPS and bacterial ghosts. These effects of the FCS may be due to the presence of LBP in the FCS. The results show that CD14 is highly relevant for the activation of mouse macrophages by bacterial cells, LPS, and lipid A. Specially defined bacterial cell wall constituents such as bacterial S-layers might act through other activation pathways.

Journal of Endotoxin Research, Vol. 4, No. 6, 431-441 (1997)
DOI: 10.1177/096805199700400607


CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    Twitter    What's this?