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(Investigative Ophthalmology and Visual Science. 2005;46:3095-3101.)
© 2005 by The Association for Research in Vision and Ophthalmology, Inc.
DOI:  10.1167/iovs.04-1365
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Potentiation of Lipopolysaccharide-Induced Chemokine and Adhesion Molecule Expression in Corneal Fibroblasts by Soluble CD14 or LPS-Binding Protein

Ken Fukuda,1 Naoki Kumagai,2 Kazutaka Yamamoto,2 Youichiro Fujitsu,2 Nobuhiko Chikamoto,2 and Teruo Nishida2

1From the Departments of Ocular Pathophysiology and 2Biomolecular Recognition and Ophthalmology, Yamaguchi University School of Medicine, Ube City, Yamaguchi, Japan.

PURPOSE. The detection of bacterial lipopolysaccharide (LPS) by human cells is facilitated by LPS-binding protein (LBP) and soluble (s)CD14. The effects of these proteins on chemokine release and adhesion molecule expression in cultured human corneal fibroblasts were examined.

METHODS. The release of chemokines into culture supernatants and the expression of the intercellular adhesion molecule (ICAM)-1 on the cell surface were determined by enzyme-linked immunosorbent assays. The intracellular abundance of chemokine and ICAM-1 mRNAs was quantitated by reverse transcription and real-time polymerase chain reaction analyses. The phosphorylation and degradation of I{kappa}B-{alpha} and the subcellular localization of NF-{kappa}B were examined by immunoblot and immunofluorescence analyses, respectively.

RESULTS. Neither sCD14 nor LBP alone affected the expression of chemokines or ICAM-1 in cultured human corneal fibroblasts. However, sCD14 or LBP enhanced the LPS-induced upregulation of ICAM-1 and the chemokines interleukin-8 and monocyte chemoattractant protein (MCP)-1 in these cells at the protein and mRNA levels. Combined stimulation with LPS and either sCD14 or LBP also induced the phosphorylation and degradation of I{kappa}B-{alpha} and the translocation of NF-{kappa}B from the cytoplasm to the nucleus of corneal fibroblasts.

CONCLUSIONS. LBP and sCD14 may play important roles in the defense of the cornea against bacterial infection, by facilitating the detection of LPS by corneal fibroblasts.




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