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Lack of MD2 expression in human corneal epithelial cells is an underlying mechanism of lipopolysaccharide (LPS) unresponsiveness

Jing Zhang,Ashok Kumar,M. Wheater,F. Yu

Published 2008 in Immunology and Cell Biology

ABSTRACT

In the present study we tested the responsiveness of human corneal epithelial cells (HCECs) and corneal fibroblasts to lipopolysaccharide (LPS), a Toll‐like receptor (TLR) 4 ligand. Purified Pseudomonas aeruginosa LPS was used to stimulate telomerase‐immortalized HCECs (HUCL) and stromal fibroblast (THK) cell lines. Exposure of cells to LPS induced a time‐dependent activation of NF‐κB in THK but not in HUCL cells, as assessed by an increase in IκB‐α phosphorylation and degradation. Concomitant with NF‐κB activation, LPS‐treated THK cells, but not HUCL cells, produced a significantly larger number of cytokines than control untreated cells. A cell surface biotinylation assay revealed that HUCL cells express TLR4 intracellularly, whereas TLR5 is expressed on the cell surface. Furthermore, reverse transcriptase‐PCR analysis revealed that HUCL and primary HCECs, in contrast to THK cells, do not express myeloid differentiation (MD)‐2. Thus, our results demonstrate that the LPS unresponsiveness of HCECs might be due to deficient expression of MD‐2, an essential component for LPS‐TLR4 signaling.

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