Calprotectin expression by gingival epithelial cells

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Abstract

Calprotectin, a heterodimer of MRP8 and MRP14 with antimicrobial properties, is found in the cytosol of neutrophils, monocytes, and human gingival keratinocytes. During inflammation of the oral mucosa, the expression of immunoreactive calprotectin appears upregulated. Given the possible cell sources, we sought to learn if epithelial cells upregulate calprotectin in response to proinflammmatory agents. First, human gingival keratinocytes were maintained in primary culture until senescence. At each passage, cells were harvested and analyzed for quantitative expression of MRP8 and MRP14 subunit mRNA by RNase protection assays and calprotectin complex by enzyme-linked immunosorbent assay. Calprotectin expression was constitutive in the primary gingival keratinocytes, but calprotectin-specific mRNA and protein tended to increase as the cells neared senescence. To test whether calprotectin expression was inducible, immortalized gingival keratinocyte cultures were treated for 2 to 4 h with lipopolysaccharide (LPS) or interleukin-1β (IL-1β). As a positive control for inducible expression, immortalized keratinocytes were incubated with phorbol myristate acetate (PMA) (50 ng/ml) for 24 h. Incubation with PMA stimulated increased expression of MRP8 and MRP14 mRNA within 2 h, peaking within 5 h. MRP8- and MRP14-specific mRNA expression by immortalized keratinocytes appeared to be unaffected by LPS or IL-1β. In contrast, LPS, IL-1β, and PMA each upregulated IL-8. These data show that calprotectin mRNA is expressed constitutively in cultured keratinocytes, while expression by immortalized cells appears to be independent of the exogenous proinflammatory agents LPS and IL-1β.

Original languageEnglish (US)
Pages (from-to)3248-3254
Number of pages7
JournalInfection and Immunity
Volume69
Issue number5
DOIs
StatePublished - May 2 2001

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Leukocyte L1 Antigen Complex
Keratinocytes
Epithelial Cells
Interleukin-1
Lipopolysaccharides
Tetradecanoylphorbol Acetate
Messenger RNA
Cell Aging
Mouth Mucosa
Ribonucleases
Interleukin-8
Cytosol
Monocytes
Neutrophils
Up-Regulation
Enzyme-Linked Immunosorbent Assay
Inflammation

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Calprotectin expression by gingival epithelial cells. / Ross, Karen F; Herzberg, Mark C.

In: Infection and Immunity, Vol. 69, No. 5, 02.05.2001, p. 3248-3254.

Research output: Contribution to journalArticle

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