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Infection and Immunity, December 2000, p. 6770-6776, Vol. 68, No. 12
0019-9567/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Involvement of CD14 and beta 2-Integrins in Activating Cells with Soluble and Particulate Lipopolysaccharides and Mannuronic Acid Polymers

Trude H. Flo,1,* Liv Ryan,1 Lars Kilaas,2 Gudmund Skjåk-Bræk,3 Robin R. Ingalls,4 Anders Sundan,1 Douglas T. Golenbock,4 and Terje Espevik1

Department of Cancer Research and Molecular Biology1 and Department of Biotechnology, Norwegian University of Science and Technology,3 and SINTEF, Division of Applied Chemistry,2 Trondheim, Norway, and Maxwell Finland Laboratory for Infectious Diseases, Department of Medicine, Boston Medical Center and Boston University School of Medicine, Boston, Massachusetts 021184

Received 26 June 2000/Returned for modification 24 August 2000/Accepted 18 September 2000

Lipopolysaccharide (LPS) and related bacterial products can be recognized by host inflammatory cells in a particulate, bacterium-bound form, as well as in various soluble, released forms. In the present study we have compared the mechanisms used by LPS, detoxified LPS (DLPS), and mannuronic acid polymers (M-polymers), in solution or covalently linked to particles, in stimulating monocytes to tumor necrosis factor (TNF) production. The addition of recombinant LPS binding protein (LBP) and/or soluble CD14 (sCD14) enhanced the production of TNF from monocytes stimulated with soluble LPS, DLPS, or M-polymer, but did not affect the response to M-polymer or DLPS attached to particles. Treatment of monocytes with antibody to CD14, CD18, or CD11b showed that CD14, but not CR3 (CD11b/CD18), mediated monocyte TNF production in response to the soluble antigens. In contrast, anti-CD14, anti-CD11b and anti-CD18 monoclonal antibodies all inhibited the response to the particulate stimuli. On the other hand, B975, a synthetic analog of Rhodobacter capsulatus lipid A, completely abrogated the monocyte TNF response induced by LPS but did not affect the TNF induction by DLPS or M-polymer, either in soluble or particulate forms. These data demonstrate that the engagement of immune receptors by bacterial products such as LPS, DLPS, and M-polymer is dependent upon the presentation form of their constituent carbohydrates, and that factors such as aggregation state, acylation, carbohydrate chain length, and solid versus liquid phase of bacterial ligands influence the mechanisms used by cells in mediating proinflammatory responses.

* Corresponding author. Mailing address: Department of Cancer Research and Molecular Biology, Norwegian University of Science and Technology, University Medical Center, N-7489 Trondheim, Norway. Phone: 47 73 59 88 41. Fax: 47 73 59 88 01. E-mail: trude.flo{at}medisin.ntnu.no.

Infection and Immunity, December 2000, p. 6770-6776, Vol. 68, No. 12
0019-9567/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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