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J. Biol. Chem., Vol. 277, Issue 38, 35489-35495, September 20, 2002
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From the The alginate capsule produced by the human
pathogen Pseudomonas aeruginosa is composed mainly of
mannuronic acid polymers (poly-M) that have immunostimulating
properties. Poly-M shares with lipopolysaccharide the ability to
stimulate cytokine production from human monocytes in a
CD14-dependent manner. In the present study we examined the
role of Toll-like receptor (TLR) 2 and TLR4 in responses to poly-M.
Blocking antibodies to TLR2 and TLR4 partly inhibited tumor necrosis
factor production induced by poly-M in human monocytes, and further
inhibition was obtained by combining the antibodies. By transiently
transfecting HEK293 cells, we found that membrane CD14 together with
either TLR2 or TLR4/MD-2 could mediate activation by poly-M.
Transfection of HEK293 cells with TLR2 and fluorescently labeled TLR4
followed by co-patching of TLR2 with an antibody revealed no
association of these molecules on the plasma membrane. However,
macrophages from the Tlr4 mutant C3H/HeJ mice and TLR4
knockout mice were completely non-responsive to poly-M, whereas the
tumor necrosis factor release from TLR2 knockout macrophages was half
of that seen with wild type cells. Taken together the results suggest
that both TLR2 and TLR4 are involved in cell activation by poly-M and
that TLR4 may be required in primary murine macrophages.
Involvement of Toll-like Receptor (TLR) 2 and TLR4 in Cell
Activation by Mannuronic Acid Polymers*
,,,,
,**
Institute of Cancer Research and Molecular
Biology and the Institute of Biotechnology, Norwegian University
of Science and Technology, 7489 Trondheim, Norway, the
§ Department of Medicine, Division of Infectious Diseases,
University of Massachusetts Medical School, Worcester, Massachusetts
01655, and the ¶ Department of Host Defense, Research Institute
for Microbial Diseases, Osaka University, Osaka 565-0871, Japan
*
This study was carried out with financial support from the
Commission of the European Communities specific research, technological development, and demonstration (RTD) program "Quality of Life and
Management of Living Resources," Grant QLK2-2000-336,
HOSPATH; the Norwegian Cancer Society; and the Norwegian Research
Council.The costs of publication of this
article were defrayed in part by the
payment of page charges. The article
must therefore be hereby marked
"advertisement" in
accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
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